Proceedings, 2018, Cell Death 2018

International Cell Death Research Congress 2018, the II edition of the conference series, was held in IZMIR at Wyndham Grand Izmir Özdilek Hotel, from November 1 to 4, 2018 for the second time in Turkey, after Izmir 2016. This conference series were organized by Cell Death Research Society of Turkey (CDRS-Turkey). [...] Full article

In the study, we aimed to show the role of autophagy which acting as a seesaw between apoptosis and necroptosis increasing and decreasing changes in certain vital organs under effects of envenomation by Mesobuthus nigrocinctus and anti-venom administration in mice. In group design, we previously classified totally 42 Albino mice into two main categories including 1st, 3th and 6th hours of envenomation and anti-venom administrated animals (at dosages of 20LD50, 30LD50 and 40LD50) before standard envenomation procudure. These were sub-divided into six groups including envenomed and anti-venom administrated mice (n = 6 at each one) were studied as well as not being evenomed and not anti-venom-administrated mice (n = 6) were used as control group (n = 6). At the end of 24 h after envenomation, all animals were sacrificed. Liver, kidneys, spleen, heart and lungs tissues were collected from all groups. After routine histopathological examination, expressions of mTOR as an autophagy activator, expressions of RIPK3 as a necroptosis activator, and caspase-3, caspase-9 as the markers of apoptotic cell death signals were evaluated by immunoperoxidase method in addition to DNA in-situ fragmentations by TUNEL method in aforementioned tissues. In envenomed groups, caspases and TUNEL reactions were low after envenomation in contrast to high RIPK3 expressions. mTOR expressions were remained at stable. In anti-venom administration groups, mTOR and RIPK3 expressions were more decreased in contrast to higher caspases exspressions and TUNEL expressions. In conclusion, we think that the scorpion envenomation drift solely the cells to necroptosis. And, under presence of anti-venom supply, the decreased mTOR expressions triggered autophagy and dependently that apoptosis was developed instead of necroptosis. We believed that this venom-antivenom administration study is a usefull role-model for better understanding of switch effect of autophagy between apoptosis and necroptosis. Full article

Flavonoids have chemo-preventive and chemotherapeutic properties against different human cancers including chronic myeloid leukemia. Quercetin and curcumin are two polyphenols with potential anti-carcinogenic and pro-apoptotic properties. We have previously demonstrated the synergistic protective effect of quercetin and curcumin on chronic myeloid leukemia cells (K562) cells. Anti-proliferative and apoptotic effects of these polyphenols were examined by apoptosis and cell viability assays. Oxidative status of the cells was analyzed by determining the level of reactive oxygen species, mitochondrial permeability and intracellular glutathione. Obtained data showed that quercetin and curcumin had beneficial and synergistic effects on K562 cells. On the basis of the above-mentioned data, herein we aimed to clarify signaling pathways involved in synergistic combination of quercetin and curcumin on K562 cells. Normal peripheral blood mononuclear cell line was used as controls. The mRNA and protein expressions of the signaling pathways were detected by Human Signal Transduction Pathway Finder-RT2 PCR Array system and Western blotting, respectively. The results of PCR array were evaluated by DAVID v6.8 and database for KEGG pathways. Our data revealed that synergistic combination of curcumin quercetin was effective on genes that were particularly related to P53, NF and TGF. We believe that our findings will lead to new research in this area and will contribute to the chronic myeloid leukemia treatment protocols. Full article

Gastric cancer (GC) is one of the most prevalent malignant types in the world and an aggressive disease with a poor 5-year survival. GC is the third greatest global cause of cancer-related deaths. It is well recognized that tumor heterogeneity, a fundamental feature of malignancy, plays an essential role in the cancer development and chemoresistance. In this study, we investigated the apoptotic effects of Pistacia vera nut skin methanol extract on human gastric cancer cell line HGC-27. The results of this study demonstrate for the first time the protective effect of Pistacia vera nut skin methanol extract on gastric carcinoma. Full article

Fas is the receptor of tumor necrosis family receptors (TNFR) and involves in apoptosis. Since discovery of T helper 17 cells (Th17) in 2005, which are defined as a new type of helper T cells, it has become clear that the dysregulated function Th17 cells and their cytokines could contribute to pathology of diseases including autoimmune diseases and cancer. There is not much known about apoptotic and survival mechanisms of Th17 cells in the literature. Therefore, the players of apoptotic cell death in Th17 cells were investigated in the study. To carry out designed experiments, venous blood were drawn from the healthy volunteers with approval from the Noninvasive Ethics Committee. Peripheral blood mononuclear cells (PBMCs) were isolated from blood with Ficoll separation method. The naïve CD4⁺ T cells were sorted from the PBMC. Sorted naive T cells were cultured under Th17 polarizing conditions. The activation, differentiation and apoptosis related molecules of cultured cells were monitored by Flow cytometry. Data showed that naive CD4⁺ T cells were activated and differentiated into Th17 cells. Activated Th17 cells were Fas positive. Activated, Fas positive Th17 cells did not underwent significant plasma membrane changes. Furthermore, it was also observed that there was not much change in the Bcl-2 protein level. Bcl-2 protein is belongs to B-cell-lymphoma-2 (Bcl-2) family proteins and is major regulator of intrinsic apoptotic pathway as promoting cell survival. In addition to that the expression of Bclx-L, is an anti-apoptotic protein, were increased in these cells. Data indicates that Th17 cells (under Th17 polarization condition) were increased expression of anti-apoptotic Bcl-2 family members. Full article

Breast cancer is the leading type of cancer in women. The majority of cells in the diagnosis of breast cancer are estrogen receptor alpha (ER α) positive and the growth of these tumors is due to estrogen. Tamoxifen is used as a supportive treatment method in breast cancer patients. Vitamin D is a group of sterols with hormone-like functions. Vitamin D is known to have anti-proliferative effect and is known to induce cell arrest and apoptosis in the G0/G1 phase in the cell cycle. This study aims to investigate the potential anti-carcinogenic effect of different concentrations of Tamoxifen and vitamin D, which are thought to have anti-proliferative effect on breast cancer cell line. In our study, combination of different concentrations of Tamoxifen and vitamin D was evaluated. As a result of the data obtained, cell cycle was studied in FACS ARIA III device. In addition, the expression of p53, BcL-2 and Bax proteins was examined in qRT PCR. In our study, low concentrations of Tamoxifen and vitamin D increased proliferation in cancer cells. An effective concentration was found for Tamoxifen. Vitamin D alone did not reduce cell proliferation but decreased cell proliferation with combination. As a result, it was found that therapies using these two agents separately decreased the proliferation of cancer cells and induced cell arrest in the G0/G1 phase in the cell cycle and changed the expressions of p53, BcL-2 and Bax proteins. Full article

In this study, the fruits and leaves of wild blueberries grown naturally in our country will be evaluated by using different infusion and boiling methods. Blueberry teas; leaves, raw fruit, dried and shredded raw fruit, fruit beans and seedless raw fruit of different infusions were used after boiling them for 1 min, 3 min, 5 min, 7 min, 10 min. Phenolic levels were determined by LC MS/MS technique. The antioxidant and activities of all products in the vitro HCT-116 colon cancer cell line were analyzed by spectrophotometric methods. MDA and TEAC were evaluated for antioxidant activity. Cytotoxicity and viability tests were performed by adding WST-8 (Water Soluble Tetrazolium Salt-8) solution. For apoptosis, TRAIL and Apaf-1 ELISA Kit were used for the activation of caspases of intrinsic and extrinsic pathways. Full article

Premature Ovarian Failure; is characterized by the dysfunction or early depletion of ovarian reserves due to follicular loss in the ovary in women under age of 40. POF is the important cause of infertility and its etiology is still not clearly understood. Investigation of cell death mechanisms (CDM) that play a role in the follicular atresia (FA) triggered by excessive loss of granulosa cells (GCs) that provide metabolic support for oocyte and follicle development in the ovary will help to understand POF etiology. It was known that apoptosis and autophagy play a role in FA. Recent studies have shown that paraptosis, associated with endoplasmic reticulum stress (ERS), also exist in FA. POF model was established in C57BL/6 female mice by CTX and it was confirmed by increased follicle stimulating hormone (FSH), luteal hormone (LH) and decreased estradiol (E2) blood levels and follicle count. According to the results of the immunohistochemistry (IHC) cell death markers were significantly more expressed than control (C) and sham (S) groups in the POF model. In addition, more apoptotic cells were observed in the POF group compared to C and S in the TUNEL analysis. In consequence of this study apoptosis and autophagy as well as paraptosis play a role in the FA leading to POF, will help to develop new treatment protocols. Full article

Neuroblastoma (NB) is the most common extracranial solid tumor in children with incidence rate 10.2 per 1 million children under 15 years old. NB has great diversity in clinical behavior while some tumors are easily curable and spontaneously regressed, the others are resistant to chemotherapies and exhibiting high relapse rates. Despite intensive usage of chemotherapies, still 5-year survival rate is about 40% for advanced stage bad prognostic patient group. Hence, there is need for more effective therapeutics and novel biomarkers for early detection of high-risk patients. In the present study we aimed to study molecular mutation profile of both good and bad prognostic NB patients with whole exome sequencing (WES). The NB patients whose risk group previously was confirmed according to TPOG-2009 protocol. 5 patients with good prognosis (low risk & intermediate with good histology) and 5 patients with bad prognosis (intermediate poor histology & high risk) were chosen. As a result, 23 common mutations were found in both prognostic group. While 7 mutations were found to be specific to bad prognostic group, 2 mutations were only observed in good prognostic group patients. The commonly mutated genes were located commonly in cell membrane and extracellular space and mostly interfering with cell signaling and cell communication pathways. Full article

The objectives of this study are to identify changes in gene expression, which involved in apoptosis pathways induced by gamma radiation, Cervical cancer cells were exposed to various doses of a single fraction of gamma radiation. After incubation for different periods, the proliferation of C-4 I and HeLa cells were investigated by MTT assay, wile morphological features were assessed by fluorescent microscopy to measure the Apoptotic Index (AI). In addition, gene expression was evaluated using micro-array molecular processes, and signalling pathway analysis were performed. Gamma irradiation inhibits proliferation of HeLa and C-4 I cells in a time- and dose- dependent manner. From our results a significant difference was observed between HeLa and C-4 I cell lines (p < 0.01), whereas, HeLa cells seemed to be radio resistant, while C-4 I cells radio-sensitive. IC50 and AI dose for C-4 I and HeLa cells were 16 Gy and 32 Gy respectively. Microarray results monitored the expression of some factors that are known apoptosis activators were up regulated by gamma radiation treatment, whereas some anti-apoptosis members were down regulated. Pathway analysis identified that significant pathways related to apoptosis, WNT, cell cycle and P53 were significantly reinforced. These results give evidence that ionized radiation directly induces anti proliferative effects by converting the expression of genes related to apoptosis and cell proliferation pathways in HeLa and C-4 I cervical cancer cells. Full article

In both healthy cells and cancer cells a number of different cellular responses influence the progress of cancer. In this study, we have investigated the effect of stress conditions on human hepatocyte cancer cells. Different concentrations of hydrogen peroxide, quercetin and N-acetyl cysteine were used to induce the acute cell stress. We have detected apoptosis, measured proteasome activity and performed a cell death array in both treated and control cells. Treatment of human hepatocyte cancer cells with hydrogen peroxide and quercetin resulted in decreased cell viability and increased apoptosis. On the other hand, proteasome activity was increased with hydrogen peroxide treatment but decreased with quercetin. N-acetyl cysteine supported cell viability and increased proteasome activity. In conclusion, cell death related genes were affected in quite different manner following hydrogen peroxide, quercetin and N-acetyl cysteine treatments. Full article

Acute myeloid leukemia (AML) is the most common form of acute leukemia and the overall 5-year survival in AML patients is 40–45% in young patients and less than 10% in elderly patients. Therefore, there is still a need to improve therapeutic approaches. After development of boron-based anticancer drugs (such as Bortezomib), boron compounds have gained attention with possible anti-leukemic feature. In this study, we evaluated anti-tumoral effects of borax pentahydrate (BP) and disodium pentaborate decahydrate (DPD) on leukemic cell line (HL-60). Cell viability was assessed by MTT assay and apoptotic effect of the compounds were evaluated by flow cytometry and cleaved PARP level detection by western blot. We observed that BP (24 mM) and DPD (6 mM) exhibit anti-leukemic effect via induction of apoptosis. Our study suggest that BP and DPD have potential anti-leukemic effect, which needs to be confirmed by further wide-spectrum studies. Full article

Breast cancer is the most common female death in women. Different studies are being done for treatment. The aim of this study was to investigate the antiproliferative, apoptotic and antioxidant effects of oleuropein and vitamin D both individually and in combination. Apoptosis is a programmed cell death pattern with genetic regulation that requires energy, allowing cells to be safely removed from the environment after performing their specified biological task. The olive tree has important biological properties and is rich in phenolic substances. These phenolic substances are mainly oleuropein. Oleuropein has many pharmacological effects such as antioxidant, antimicrobial, antiviral, anticancer, antiinflammatory, antiageing. Vitamin D is a hormone the kidneys produce that controls blood calcium concentration and impacts the immune system. Full article

Hematopoetic Stem Cell Transplantation (HSCT) has benefical effects for the treatment of leukemia but one of the adverse effect is Graft versus Host Disease (GVHD) that causes high mortality and morbidity. Mesenchymal Stem Cells (MSCs) are one of the promising tools which are used for the treatment of steroid refractory GVHD. The success of cellular therapies depends on so many factors associated with the donor and host characteristics. We designed a study concerning that blood group reagents at different titres can affect the survival of MSCS in blood as they were injected intravenously when used for the treatment of GVHD. We concluded that proliferation rates of MSCs are decreasing in relation with the high reagent titres. Flow-cytometric analysis also revealed that high titers of reagent in culture medium could lead to the induction of apoptotic pathways in MSCs. For individualized treatment strategies, donor and host characteristics must be taken in to consideration in details and experimental studies in cell culture laboratories could be essential to gain insight into clinical trials. Full article

Although the endothelial dysfunction is related with tumor development, there is no consensus on the suppressive or supportive effect on tumor growth. The goal of the present study was to evaluate endothelial dysfunction related factors in animal breast cancer model that was developed by administrating endothelial nitric oxide synthase blocking agent, Nitro-L-arginine methyl ester hydrochloride (L-NAME). Endothelial dysfunction related main factors such as nitric oxide synthase, interleukin-6, vascular endothelial growth factor receptor-2 and vascular endothelial cadherin were investigated by immunohistochemically in tumor and carotid artery tissues. In tumor tissues apoptosis was determined by TUNEL assay. The level of endothelin-1 in blood was measured by ELISA. İntima-media thickness of carotid artery was evaluated with Doppler-USG measurements. As a result, in this study it was shown that vascular endothelial growth factor receptor-2, endothelin-1, endothelial nitric oxide synthase, interleukin-6, vascular endothelial cadherin and E-selectin molecules play a role in reducing breast tumor growth based on endothelial dysfunction. Full article

In this study, the wound healing effect of ozonated hazelnut oil was investigated on 54 male Sprague-Dawley rats. 7 experimental groups were designed. A circular area with a diameter of approximately 2 cm on dorsal surface was drawn using a coin and full thickness of the marked area was cut approximately 3 cm away from the ears by sterile scissors and forceps. All oils were daily applied on wound surface of experimental animals in volume of 0.2 mLduring 15 days. In this study, statistical analysis of the data was performed using the SPSS 22.0 package program. Wounds of all experimental animals were photographed by a digital camera on days 0, 3, 6, 12 and 15 after wounding. Ozonated oil treated wounds had significantly higher than the other groups on the day 3 after wounding. There is a quantitatively healing is determined on 3 day on rats. It has been observed in the stability period that ozonated oils must be stored at a temperature below 4 °C. Vitamin E should be added as an additive in the medical use of ozonated hazelnut oil in wound healing. Full article

Recent studies have shown that cancer cells can deceive phagocytosing macrophage cells through the CD47 protein which gives the message “don’t eat me” or “don’t kill me” to immune components. The efficacy of anti-CD47 treatment approach was shown in cancers such as, non-small cell lung cancer, non-Hodgkin lymphoma, ovarian cancer, and breast cancer. The studies on the immunobiology of neuroblastoma has increased as monoclonal antibody based immunotherapy has shown to be effective in high-risk patients such as anti disialoganglioside. Therefore, the aim of this study was to evaluate the levels of CD47 protein expression among neuroblastoma patients with different risk groups and genetic alterations. This study included paraffin-embedded tumor tissues of 66 neuroblastoma patients (28 girls, 38 boys) with an age range of 0.5 to 108 months with a mean value of 24.9 (±23.5). According to risk classifications 21 were at low risk (31, 8%), 24 were at intermediate risk (36, 4%) and 19 were at high-risk (28, 8%) groups. These samples were evaluated for MYCN amplification, 1p36 LOH, 11q23 deletion and 17q25 gain by real-time PCR. In addition, CD47 expression status (positive or negative) was detected by immunohistochemical analysis. All data was analyzed with Chi-Square and Mann-Whitney U non-parametric tests within SPSS program, version 22.0. p value lower than 0, 5 was found statistically significant. According to the results, patients at low risk did not express CD47, while patients at high-risk group were mostly expressing CD47 (p = 0.049). MYCN amplification positive patients were expressing CD47 protein (p = 0.046). Patients without 17q25 gain were found to be expressing CD47 protein (p = 0.006). In addition, CD47 expression was increasing as age was getting higher in terms of months (p = 0.018). The findings of this study suggest that positive expression pattern of CD47 may be a poor prognostic biomarker especially in high risk 17q gain negative neuroblastoma patients. Full article

Psoriasis is a chronic and relapsing skin disease that affects approximately 2–3% of the world population and is considered as an inflammatory disease. Defective keratinocyte proliferation and differentiation programs, aberrant immune responses are the major factors that have been implicated in its pathogenesis. Thick scaly plaques, which are psoriasis characteristic formed by the abnormal maturation of keratinocytes and epidermal hyperplasia. The aim of this study is to establish the initial stage of psoriasis model by LPS (lipopolysaccharide) treatment to HaCaT (transformed keratinocyte) cells. MTT assay of LPS was performed to hallmark the initial stage of psoriasis. ELISA assay was performed to determine the cytokine release as response to LPS for psoriasis initial stage. According to the MTT assay results 200 ng/mL used as the test dose and 8µg/mL LPS determined as the toxic dose at 72th hour. LL-37 was the first cytokine whose expression increased in the formation of the immune response. Besides, TNF-α release occurs soon after. The presence of immune system cells is needed to maintain IL-17 and IL-1β release. The cytokine release at the 72th hour decreased to control level due to the lack of immune cells in the culture medium. Full article

Infection and chronic inflammation contribute to about 1 in 4 of all cancer cases. Emerging evidence suggests that the host factors in the tumor microenvironment may interact with underlying inflammatory prostate cancer cells to make them aggressive. In this study, we hypothesized that soluble factors secreted by immune cells activated by inflammation can induce EMT in prostate cancer and thus promote metastasis. We identify that macrophage-secreted cytokines including TNFα act as mediators for potentiating LNCaP cell proliferation in migration assay. Hence, our data indicate a mechanistic insight of how inflammation may contribute to development of prostatic disease at an early stage through increasing cell proliferation and metastasis of LNCaP cells. Full article

IL-17 producing T helper 17 (Th17) cells are identified as a distinct subset of CD4⁺ T helper cells. They play a role in immune response. Abnormal regulation of Th17 cells can play a role in different type of pathologies such autoimmune diseases and cancer. The apoptotic and survival mechanisms of Th17 cells are not well known in different type of diseases. Therefore, the aim of the study was to investigate Bcl-2 family proteins to understand the regulation network of apoptosis in human Th17 cells. To do that, Peripheral blood (PB) were drawn from the healthy volunteers. Peripheral Blood Mononuclear Cells (PBMCs) were isolated from blood by Ficoll gradient isolation method. The naïve CD4⁺ T cells were isolated from PBMC. Sorted naive CD4⁺ T cells were cultured under Th17 polarizing conditions. Th17 cells were characterized by Flow cytometry. Cell lysates were obtained from negative controls and Th17 cells. Bcl-2 family members (Bik, BID and Puma) in Th17 cells were detected by western blot. Data showed that naive T cells were differentiated into Th17 cells. Then, cell lysate of this cells were used for western blot experiments. In the Th17 cell lysate, BH3 family members Bik and Puma were not detectable but Mcl-1 expression was increased. Overall data indicated that the pro-apoptotic BH3-only subgroup proteins Bik and Puma was not detectable, however anti-apoptotic Mcl-1 protein expression was increased. Full article

The aim of this study was to investigate the anticancer activities of five chloroform extracts prepared from the aerial parts of Centeura kilaea Boiss. (CKC), C. cuneifolia Sm. (CCC), C. salicifolia M.Bieb. ex Willd. (CSAC), C. stenolepis Kerner (CSC1 and CSC2) against hepatocellular cancer HepG2 cell line. At the same time, antioxidant and anti-inflammatory activities of the extracts were examined. Anticancer, anti-inflammatory, antioxidant activities of extracts were carried out by MTT, lipoxygenase, DPPH-ABTS methods, respectively. CCC, CSAC and CKC have the highest anticancer activity when compared to other extracts. Similarly, CSAC and CCC had the best antioxidant capacity, while CSAC and CKC showed strong anti-lipoxygenase activity. Among all the tested extracts, the highest amounts of total phenolic were found in the CSAC. The study revealed that CSAC and CCC displays remarkable antioxidant, anticancer and anti-inflammatory activity. Full article

Colchicum pusillum Sieber is a plant from Colchicaceae family that particularly rich in tropolonic alkaloids, colchicine. The aim of the study is to determine the apoptosis induction effect of Colchicum pusillum extract in Colo-741, metastatic colon cancer cells and if so, to determine by which apoptotic pathway. Colchicum pusillum was collected and extracted with ethanol. Apoptotic activities of Colchicum pusillum were investigated by immunocytochemistry using antibodies directed against to caspase-3, cytochrome-c and FasLigand (FasL) in Colo-741 cells. Immunocytochemical staining results stated that Colchicum pusillum extract at 20 µg/mL dose, increased caspase-3 and FasL activity significantly but was not able to activate cytochrome-c activity. The results suggested that Colchicum pusillum extract induced apoptosis in metastatic colon cancer cells via extrinsic apoptotic pathway. Full article

Helicobacter pylori, a helix-shaped gram-negative microaerophilic bacteria has been classified as a human carcinogen (Group I carcinogen) by the International Agency for Research on Cancer (IARC) on the basis of numerous animal and epidemiological studies. Chronic Helicobacter pylori infection can cause lead to high levels of intracellular reactive oxygen species (ROS) and genomic instability both directly by genetic and/or epigenetic pathways. It is known that high levels of intracellular ROS can trigger apoptosis and induce DSBs. In this study, we aimed to investigate intracellular ROS levels and apoptotic effects of Helicobacter pylori infection on AGS cells in the presence of a DSB repair inhibitor [8-(4-dibenzothienyl)-2-(4-morpholinyl)-4H-1-benzopyran-4-one, Nu7441]. After incubation of AGS cells with different multiplicities of infection (MOIs; 25, 50, 75, 100, 200 and 400), we observed that Helicobacter pylori causes MOI-dependent cytotoxicity and intracellular ROS levels significantly increased in all study groups vs. control. Both caspase 3 and caspase 8 levels were higher all of the study group when compared to control group. In HP+Nu7441 group, caspase 8 levels 3.5 times higher vs. control; indicating unrepaired DSBs in Helicobacter pylori infection can highly be induce apoptosis in AGS cells. Other cell death mechanisms, including autophagy, should be studies in order to fully understand the cytotoxicity mechanisms of the bacterium. Moreover, we can state that in organisms with unrepaired DSBs or with high DSB levels, Helicobacter pylori can cause more oppressive adverse effects, the importance of which should be elucidated with mechanistic studies. Full article

Metoclopramide, used as an anti-emetic drug in clinical practice, has recently also begun being used to establish hyperprolactinemic effects in the breastfeeding. The purpose of this study was to investigate the potential side-effects of metoclopramide applied in the lactation period in the central nervous system of infant rats. 18 female albino Wistar rats that had just given birth were divided into 3 groups together with their pups: Healthy controls, low-dose metoclopramide (10 mg/kg, twice per day i.p.) and a high-dose metoclopramide group (45 mg/kg, twice per day i.p.). Brain tissues from 6 pups from each mother were harvested at the end of the 21st day. Immunohistochemical technique was performed using dopamine D2 receptor (DRD2), brain derived neurotrophic factor (BDNF) and neural growth factor (NGF), markers of extrapyramidal reaction in the brain, as signal molecules. Based on immunohistochemical results, DRD2 expression decreased only in the external pyramidal layer neurons in the high-dose infant group. Strong BDNF reaction was determined in pyramidal neurons in all layers in the control infant group, and decreased reaction was observed in the high- and low-dose groups. No significant difference was observed in NGF expression between the three groups. Since high-dose metoclopramide caused a decrease in DRD2 expression in the external pyramidal layer in the prefrontal cortex, and since both high and low doses reduced BDNF expression, care needs to be taken with the use of metoclopramide in the lactation period due to the possibility of extrapyramidal reactions in infants. Full article

Nicotine, is an alkaloid compound consisting of pyridine and pyrolidine ring. Its closed formula is C10H4N2. During smoking peak plasma concentration changes from 25 to 50 ng/mL. Its half-life is 1–2 h. Nicotine is metabolized primarily in the liver and excreted by the kidneys. The plasma concentration of the nicotine metabolite cotinine is 10 times more than nicotine and its half-life is longer around 15 to 20 h. Cotinine can be found in both amniotic fluid and umbilical cord blood given that it passes across the placental barrier. Adverse effects of nicotine and its metabolites on the fetus are suggested but have not been proven by scientific explanations till now. Sialic acid (Sia) is a modified nine-carbon sugar. They are located on the last end of the glycan chains located in the glycoconjugate structures. They organize a wide range of relationships between cells and their environment such as cellular recognition, adhesion, transmission, differentiation and aging. The aim of this study is to determine the possible changes in the sialic acid levels and types in the plasma after different levels of nicotine applied to Swiss Albino rats in order to assess the effect of long-term per oral nicotine administration. Full article

Ewing sarcoma is a bone and soft tissue tumor either neuroectodermal or mesenchymal originated and affecting children and adolescents. In the present study, we aimed to find out prognostic and predictive biomarkers for Ewing sarcoma. Hence, we examined the copy number alterations (and related possible genes) among ten Ewing sarcoma patient samples and possible associations with the clinical outcome. DNA extraction from formalin fixed paraffin embedded archive tissues was performed. Whole genome Comparative Genomic Hybridization (CGH) was performed by NimbleGen and recorded as single Panel Rainbow through chromosomes 1–22, X and Y. Data was interpreted by SignalMap software and genetic regions matching the deletion or amplification loci were recorded. The mean age of the patients was 8.6 years. Three of the cases were male, while seven were female. According to CGH analysis, the most common DNA copy number alterations were found in SLIT-ROBO Rho GTPase activating protein (srGAP2), RANBP2 like GRIP domain (RGPD5), nephrocystin 1 (NPHP1), GTF2I repeat domain containing 2 (GTF2IRD2), pyridoxal dependent decarboxylase domain containing 1 (PXDC1), which were found down-regulated among 7 of 10 patients. In conclusion, in our dataset the copy number alterations are mostly found in genes related with cytoskeletal elements, migration and protein trafficking among our patient group. Gene functional studies are required for better understanding the role of these genes in Ewing Sarcoma pathogenesis Full article

Anti-apoptotic Bcl-2 and proapoptotic Bax genes are the most significant genes that are involved in the regulation of apoptosis. Abnormal apoptotic activity in preeclampsia and gestational diabetes is caused by dysregulation of these genes. In this study; we examined Bcl-2 and Bax protein expressions using immunohistochemical techniques in human placental tissue samples from cases of gestational diabetes (n: 20) and preeclampsia (n: 20). It was observed that Bax expression showed positive reaction compared to Bcl-2 expression so; Bax protein was thought to be an effective marker in determining apoptotic changes in placentas with gestational diabetes and preeclampsia. Full article

Beta-thalassemiais one of most common autosomal recessive hematologic disorder. Alterations in the lipid profile have been reported consistent with β-thalassemia, but the pathogenesis is not clear. Genetics variations within in both the coding and noncoding regions of the PPAR-alfa gene locus contribute to dyslipidemias and imply hematologic risks. Therefore, this study is designed to investigate the lipid profiles in patients with β-thalassemia major and intermediate and examine the possible association with dyslipidemias causing susceptibility PPAR-α gene variants in the population of Turkish Cypriots, where the prevalence of thalassemia and carriers are very high. Our results showed the strong association with the PPAR-α rs4253778 CC genotype and triglyceride in beta-thalassemia major and intermediate patient. It is tempting to speculate here that the PPAR-α gene rs4253778 CC genotype could possibly impair triglyceride metabolism more effectively in beta thalassemia major and intermediate subjects. Overall, the results from our study establish the homozygous wild-type C allele at the PPAR-α rs4253778 locus as a genetic risk factor with a riglyceride-elevating effect in a population of Turkish Cypriot β-thalassemia patients. Full article

Although cancer research undergone a rapid expansion, there is no potential cure and the disease remains one of the leading causes of mortality worldwide. Breast cancer continues as the female malignancy and a major cause of death in middle-aged women. Calmodulin can interact with large number of different target proteins and modulate their activity in different ways. Calmodulin antagonists have been reported to induce apoptosis and inhibit tumor cell growth. However, the potential effects of these target agents on various cancer within the molecular levels are poorly understood. The main objective of this study is to evaluate the potency of a number of calmodulin antagonists on the growth of human breast cancer cell lines. In addition, preliminary trials were carried out to find the role of these agents within the molecular levels. The finding indicates that Calmodulin antagonists might be the key for future breast cancer treatment strategy. Full article

Mutations in the gene for fibroblast growth factor receptor 3 (FGFR3) are implicated in achondroplasia, an autosomal-dominant form of short-limbed dwarfism. The present study involves a combination of clinical exome sequencing, targeted resequencing and protein modeling methods to decipher the pathobiology of achondroplasia with psychomotor delay in a two-year-old child. Accordingly, the resulting genetic information establishes the frequent FGFR3 c.1138G > A (p.G380R) mutation as the single hit causing pediatric achondroplasia with psychomotor delay, while the predicted model stresses the importance of a phenylalanyl residue (F384) in enhancing the dimerization potential of the receptor’s transmembrane domain via a cation‒π interaction with the newly introduced arginyl residue. Overall, the likely involvement of FGFR3^G380R in psychomotor delay calls for comprehensive clinical assessment in achondroplastic children, although the precise mechanism by which the mutant receptor results in the development of neurological manifestations awaits further investigation. Full article

After physical or surgical damage of corneal epithelium, most of analgesic drugs, like non-selective opioid agonists and non-steroid anti-inflammatory drugs, cannot be used because of their negative effects on wound healing process. Biphalin is selective µ and Δ opioid receptor agonist which has proven analgesic effects on rodents. Our purpose of study is finding effects of biphalin on wound healing of corneal epithelium. We used primary culture of human corneal epithelial cells (HCECs) for examining effects of biphalin on wound healing. Firstly, we measured toxicity of Biphalin in various concentrations with MTT assay and we showed biphalin has no toxic effects on HCECs in lower concentrations than 100 µM in various incubation times. After MTT assay, we administered 1 µM and 10 µM biphalin at in vitro scratch assay of HCECs, biphalin increased wound closure process significantly at 1 µM concentration (p < 0.05). Then we tested effects of biphalin on cell migration and proliferation separately. Bifalin increased migration of HCECs significantly (p < 0.01) at transwell migration assay. But we did not observe any significant difference between groups in Ki67 proliferation assay. In all these experiments, we also used naloxone to inhibiting effects of biphalin. In biphalin plus naloxone groups, effects of biphalin decrease partially. Our study results suggest, biphalin has positive effects on epithelial wound healing via opioid receptors. This effect because of increased migration of HCECs under influence of biphalin. With these findings, we propose biphalin as a new analgesic agent for post-surgical and post-traumatic care of corneal epithelial wounds. Full article

Exposure to 3,5-dimethylaminophenol (3,5-DMAP), the metabolite of the 3-5-dimethylaniline, was shown to cause high levels of oxidative stress in different cells. However, we have shown that this alkylaniline metabolite was non-mutagenic to different strains of Salmonella typhimurium in Ames test and also was found to be not mutagenic to CHO cells in HPRT test. Concerning all the available data, we aimed to observe whether this metabolite may have anti-carcinogenic potential in human non-small cell lung cancer line (A549 cells). 3,5-DMAP caused a dose-dependent increase in cytotoxicity and generation of superoxide (O₂-.) and reactive oxygen species (ROS). 3,5-DMAP did not produce significant cytotoxicity to human lung fibroblasts even at very high concentrations; however showed higher cytotoxic effect on A549 lung cancer cells at the same concentrations. 3,5-DMAP also led to molecular events, like increases in apoptotic markers (i.e., p53, Bad, Bax and cytochrome and decreases anti-apoptotic proteins (Bcl-2). Furthermore, 3,5-DMAP provided significant decreases in cell viability of A549 cells and eventually inhibited growth of A549 cells in an in vivo mouse model. Tumor sections showed that 3,5-DMAP down-regulated c-Myc expression but up-regulated p53 and cytochrome c, all of which might result in tumor growth arrest. In conclusion, our findings demonstrate 3,5-DMAP is not mutagenic to Salmonella typhimurium and CHO cells; toxic to A549 cells and therefore may have anti-cancer properties, the importance of which should be elucidated with further mechanistic studies. Full article

Cisplatin is prevalently used as a chemotherapeutic agent for testis cancer, ovarian cancer, bladder cancer, brain tumors and many cancers such as those. Infertility and sterility may emerge, after treatment with chemotherapeutic agents, depending on patient characteristics and the dose of chemotherapeutic agent. Fucoidan is a sulfated polysaccharide, which is derived from brown seaweeds. It is first isolated in 1913 from marine brown algae by Henrik Kylin. Fucoidan is especially found in the cell-wall of marine brown algae. Many effects of fucoidan such as anti-inflammatory, anti-viral, anti-complement, anti-cancer, anti-oxidant and anti-coagulant effects have been revealed by many studies in the literature. Furthermore, some studies suggest that fucoidan may enhance the antineoplastic effects of anticancer drugs. Besides beneficial effects of fucoidan, no toxic or adverse effect has been shown to date. In this study, we aimed to investigate the effects of fucoidan on testicular tissue and its effects on cisplatin induced testicular damage when used concomitantly with cisplatin. Full article

The studies on the relationship between inflammation and cancer progression have been mostly carried out with monolayer cell cultures in vitro, which can be insufficient to mimic tumor tissue. Here, we established a three-dimensional (3D) cell culture model of inflammatory microenvironment for prostate cancer cells to better evaluate the role of inflammation in prostate carcinogenesis. Formation of the cell spheroids has been achieved for LNCaP, Du145, LNCaP-104r2 prostate cancer cell lines but not for RWPE1 normal prostate epithelial cell and PC3 by using 3D Petri Dish^®. We also showed that cells in inflammatory conditioned media might have a different response based on the culturing method. Overall, we are suggesting that 3D cell culture model can be a useful tool to study molecular alterations on proliferation and migration/invasion of tumor cells related to inflammation. Full article

In this work, we focused on the development of a novel point of care (POC) paper-based analytical microfluidic chip. The system has been developed for paper-based extraction, non-enzymatic amplification and electroanalytical detection of human papillomavirus (HPV). The device comprises paper extraction support material as well as paper-based detection tool. Herein, a special DNA modification method has been utilized to allow non-enzymatic amplification by using microbeads and silver nanoparticles labeled primers. The device is capable of extracting more than 10–100 copies per mL of DNA approximately in 15 min along with a single step extraction process from patient samples. In addition, it is able to detect low concentrations taking just less than 10 min with high selectivity to HPV kinds of 16&18. It only takes the low-cost point of care device less than 40 min with a low limit of detection (LOD) to complete the whole process. Full article

Metal oxides have been widely used in various applications such as biomedical, commercial and environmental, due to their unique physicochemical properties. As the use of metal oxides increase worldwide, their exposure to the living systems also increases. It is therefore necessary to understand their potential harmful effects on human and environment health. In this study, dermal fibroblasts were exposed to bulk zinc oxide (0.1, 1, 10, 50, and 100 μg/mL) for 6 and 48 h. After exposure, changes in cell viability, morphology, membrane damage and zinc oxide uptake were investigated. The response of dermal fibroblasts exposed to different concentrations of bulk zinc oxide was monitored in real-time using an impedance-based assay system. Results demonstrated that zinc oxide at 50 and 100 μg/mL showed significant toxic effects compared to the control cell cultures. Full article

Apoptosis, a programmed cell death, has a vital role in various cellular processes. Apoptotic cells exhibit morphological and biochemical changes, detected by a variety of assays (caspases, mitochondrial dyes, DNA laddering). Flow cytometry is a powerful tool for detection of apoptotic cell death and allows information about the cell size and molecules associated with cell-bound antibodies. Recently, Fourier transform infrared (FTIR) spectroscopy as rapid and low-cost tool has been extensively used for cellular studies, providing information on cellular structures. The aim of this study was to detect early apoptosis and obtain further insights into the capability of FTIR spectroscopy, comparing the results with flow cytometry. In this study, apoptotic cell death was induced in human Jurkat T cells with Camptothecin (CPT), a DNA topoisomerase I inhibitor. Cells were cultured with 4µM CPT in RPMI (with 5% FCS) for 24 h. Immunoflourescence labeling for multicolor flow cytometry was accomplished with Annexin V concomitantly with 7-AAD. The same cells were also analyzed with ATR-FTIR spectroscopy. Flow cytometry data represents that the cells are Annexin V positive but 7AAD negative. This indicates that cells are in the early apoptotic stage, only externalization of phosphatidylserine exists on the plasma membrane. FTIR data reveals that membrane phospholipids and proteins undergo changes; fatty acid acyl chains are disordered and increased in mobility after treatment, which result from the early apoptosis process after CPT-treatment, confirmed by the flow cytometry. A combined study of flow cytometry and FTIR spectroscopy for analysis of apoptosis in human T cells exhibited compatible and complementary results. Existence of biophysical and biochemical changes in T cells after treatment were also demonstrated. Full article

Parkinson’s disease (PD) is the second most common neurodegenerative disease with an estimation of 10 million people living with the disease and it is increasing in prevalence every year. Familial cases of PD are source of valuable information to determine genetical risk factors yet sporadic cases can emerge from distinct mechanisms so, identifying common pathways of familial and sporadic cases of PD may provide worthwhile insights to determine underlying mechanisms through the progression. LRRK2 mutations are the most common indicators of both sporadic and familial cases of PD and α-synuclein aggregation is one of the hallmarks of both cases of PD as well as in other synucleinopathies. As in the case of most neurological diseases, human studies addressing the molecular basis of pathology are generally restricted to post-mortem materials. For this reason, cell culture systems and animal models are widely used. There are two main approaches for modelling PD: genetically constructed PD models and neuroxin-based models. In this study, we aim to construct and compare both approaches by overexpressing wild-type (WT) and A53T mutant α-synuclein and treating cells with well-known neurotoxin 6-hydroxidopamine (6-OHDA) using dopaminergic human neuroblastoma SH-SY5Y cell line. Our findings suggest that WT or A53T α-synuclein overexpression by itself is not sufficient to cause significant toxicity in SH-SY5Y cells in the presented time scale. As expected, 6-OHDA treatment caused toxicity with an IC50 value of ~100 µM. In addition, 6-OHDA treatment causes 3- and 2.5-fold increase in SNCA and LRRK2 expression respectively. Full article

Flow cytometry (FC) is a powerful and reliable system for cell death studies. It allows to study the molecular changes in both the surface and cytoplasmic part of the cells. Depending on the laser range of flow cytometry, it is possible to collect a large number of information about a single cell by combining multiple labeling strategies. With these assets, flow cytometry allows scientists to accelerate their research. Flow cytometry is also widely used in clinical studies. Therefore, we used this powerful tool to study human T helper 17 (Th17) cell apoptosis. Newly discovered Th17 cells are important players of immune response regulation. They are also involved in different types of pathologies including autoimmune diseases and cancer. There are intensive data in the literature about molecules that are involved in Th17 differentiation signaling networks, but the apoptotic and survival molecular mechanisms of this cells are not fully understood yet. Therefore, apoptosis of Th17 cells were measured by Flow cytometry. Healthy human subjects have been invited to study with the informed consent which is approved by the ethics committee. Human Peripheral Blood Mononuclear Cells (PBMCs) were isolated from peripheral blood of healthy volunteers. Phenotypically characterized and sorted naïve CD4+ T cells from PBMC were cultured under Th17 polarizing conditions. IL-17, IL-22 or CCR6 molecules were used to monitor Th17 cells. Apoptotic cell death of Th17 cells were measured by plasma membrane changes and DNA fragmentation. During differentiation stages, plasma membrane changes were monitored by Annexin V concomitantly with 7AAD. At day 7, there were Annexin V positive cells in Th17 cells. Apoptotic cell death occurs through sequential events including caspase activation and DNA fragmentation. DNA fragmentation data showed that Th17 cells were not apoptotic compared to negative control cultures. This finding suggested that survival molecules of Th17 cells interferes with this apoptotic process. Full article

Irisin is a protein, which is encoded by the FNDC5 gene. Irisin secreted as a hormone secret via Fibronectin type III domain-containing protein 5. Irisin is always provide beneficial effect of exercise on cellular metabolism as well as obesity and metabolic diseases. In addition, it plays a role in cancer. Therefore, in this study, aimed that to an overview of clinical research and explain the molecular mechanisms related to cancer of Irisin. This review will be providing a systematic analysis of recent work on Irisin and future work will provide a different perspective. Full article

PTEN is inactivated in a subset of lung cancer; therefore, we investigated the involvement of PTEN inactivation in invasiveness of lung cancer cells. AKT at Ser473 was phosphorylated in several lung cancer cell lines with loss of PTEN expression. Therefore, we created a tetracycline inducible expression system of wild-type PTEN (PTEN-WT) as well as catalytically (PTEN-G129R) and lipid phosphatase (PTEN-G129E) inactive PTEN mutants using the PC14, lung adenocarcinoma cell lines, in which endogenous PTEN expression was not detected and AKT at Ser473 was phosphorylated by Western blot analysis. These results suggest that the PTEN gene cannot induce apoptosis alone, but apoptosis can be achieved by the suppression of other pathways. The absence of PTEN expression gives an invasive and metastatic phenotype to cancer cells, and also while staging of cancer cells that allows the transition to a further stage of cancer. Full article

Diabetes mellitus (DM) is a complex disease characterized by absolute insulin deficiency or resistance leading to hyperglycemia. Type 2 DM is caused by development of cellular resistance to insulin combined with insufficient insulin production. Type 2 DM is a common disorder causing hyperglycemia, impaired homeostatic process, inhibition of inflammatory response, generation of reactive oxygen species. Apoptosis is a crucial process that influence normal development and tissue homeostasis. Apoptosis is known to be reduced in diabetics. Apoptotic proteins such as bcl-2 and Bax proteins regulate apoptosis. Bcl-2 serves as anti-apoptotic protein while Bax is pro-apoptotic. The balance between these apoptotic proteins determine the apoptotic process. The aim of this study was to examine the expression of apoptotic proteins Bax and Bcl-2 with immunohistochemical methods in the blood samples of type 2 diabetic patients. Full article

Endometrial cancer is the most common cancer of the female reproductive system. Perifosine has shown anti-tumor activity in a variety of cancers by inhibition of AKT phosphorylation. The active metabolite of vitamin D (1,25(OH)2D3) has anti-proliferative and pro-apoptotic properties and is recognized as an agent with the great potential for cancer intervention. Combination treatment with specific agents has been widely used as targeted therapy for cancer. In this study, we aimed to investigate the anti-apoptotic effect of varying concentrations of perifosine and vitamin D on endometrial cancer cell line (HEC1A). The expression levels of P53, BCL2 and BAX were assessed in cultured HEC1A cells at 10 and 30 μM concentrations of perifosine, 50 and 200 nM of vitamin D and combination groups using qRT PCR. Apoptosis evaluation in HEC-1A cells was performed using flow cytometry after 48 and 72 h of treatment with perifosine, vitamin D and combinations of both. According to the obtained data, treatment with different doses of perifosine and vitamin D significantly induced apoptosis in a time and dose-dependent manner in HEC-1A cells. The mRNA expression levels of P53, BAX, BCL2 in HEC-1A cells were found to be significant in the experimental groups as compared with the control. Perifosine displayed substantial anti-tumor activity in HEC-1A. These effects were increased with vitamin D. Therefore, perifosine and vitamin D combinations should be further evaluated in clinical studies in endometrial cancer. Full article

Breast cancer is the most common cancer among women and in order to create alternative treatments different types of in vivo and in vitro studies have used various plant-based therapeutic agents. Humic acid (HA) induces apoptosis and has various pharmacological properties including anti-inflammatory and anti-proliferative effects. In our study, we examined the cytotoxic effects of HA at concentrations of 5, 10, 20, 50 and 100 μg/mL in human breast adenocarcinoma MCF-7 cell line for 24 and 48 h. By using MTT method, it has been found out that HA 100 g/mL had cytotoxic effect on human breast adenocarcinoma MCF-7 cell line at both 24 and 48 h; also found out that the effective dose of HA at the same time (24 and 48 h) was 50 μg/mL. The results of our study will shed light on the development of alternative therapeutic approaches in the treatment of cancer by evaluating the cytotoxic effect of HA. Full article

Opuntia ficus indica L. fruit (cactus pear) seed oil is used in traditional and complementary therapies for its numerous health benefits. The aim of this study was to analyse of the fatty acid content and apoptotic induction effects of spiny and thornless Opuntia ficus indica L. seed (CPS) oils. Spiny and thornless Opuntia ficus indica L. seed oils obtained by supercritical CO₂ extraction method and analyzed by GC-MS. Different concentrations of almond oils were incubated for 24 h and 48 h with Colo-320 and Colo-741 cells. Cell growth and cytotoxicity were measured by MTT assays. TUNEL assay was used to detect DNA fragmentation in both cell lines. Linoleic acid was dominant fatty acid followed by oleic acid, palmitic acid and elaidic acid in both type of seed oil. In MTT, spiny CPS oil was found to be active against Colo-320 and Colo-741 cells with 1:16 dilution for 48 h. Also, 1:8 and 1:16 dilutions of thornless CPS oil showed significant reduction in the number of viable cells in Colo-320 and Colo-741 cells, respectively. The number of TUNEL positive cells were significantly higher in Colo-320 cells treated with thornless CPS when compare with control group (p < 0.05).We conclude that thornless CPS oil may have anticancer effect on primer colon adenocarcinoma cell lines. The effect can be explained by inducing apoptosis. Thus, they could be a potential novel therapeutic agent in colon cancer therapy. Full article

Breast cancer treatments continue to be investigated with supported by new treatment methods. Melatonin is a hormone that can be effective in the treatment of breast cancer due to its anti-oxidant effect. Melatonin had previously shown to inhibit proliferation of cancer cells. In this study, we aimed to determine the effect of melatonin on the proliferation of metastatic breast cancer cells in comparison to doxorubicin, a well-known chemotherapeutic agent. Doxorubicin inhibited proliferation of metastatic breast cancer cells while melatonin has no effect. We are currently examining the effects of melatonin and doxorubicin combination therapy on metastatic breast cancer cells. Full article

Autocrine growth hormone (GH) induced cell proliferation, invasion-metastasis and drug resistance in breast cancer cells. Curcumin has an apoptotic effect on colon, melanoma, cervix, and breast cancer cells. Autophagy and endoplasmic reticulum (ER) stress are essential cellular processes activated under nutrient deprivation, pathogen infection and drug exposure. Our aim in this study is to investigate the time-dependent effect of curcumin on ER stress and autophagy and potential increase of curcumin efficiency by bafilomycin treatment. Autocrine GH expression triggered resistant profile against curcumin-induced cell viability loss in MCF-7 cells. However, this effect was prevented by the time-dependent manner in MCF-7 cells. In GH+ breast cancer cells bafilomycin increase curcumin-induced cell viability loss by MTT cell viability assay. In conclusion, autocrine GH-triggered curcumin resistance was overcome by autophagy inhibition condition by bafilomycin treatment in a dose-dependent manner in MCF-7 GH+ breast cancer cells. Full article

Curcumin, a plant derived natural compound, has anti-oxidant, anti-proliferative and apoptotic effect on various cancer cells such as prostate, colon and breast cancer. Autocrine growth hormone (GH) expression induced breast cancer invasion-metastasis has been reported in vivo and in vitro cancer models. Autophagy is a vesicule-mediated clearance mechanism and one of the handicap against drug-induced apoptotic cell death. In this study, our aim was to investigate the molecular machinery of curcumin induced apoptotic cell death under autophagy inhibition conditions in autocrine GH expressing MDA-MB-231 and T47D breast cancer cells. Although autocrine GH induced curcumin resistance, this effect was slightly prevented by time-dependent curcumin treatment in MDA-MB-231 and T47D breast cancer cells. In addition, curcumin induced autophagy vacuole formation was determined by acridine orange staining in MDA-MB-231 and T47D wt/GH+ breast cancer cells. Moreover, curcumin triggered autophagy through upregulating Beclin-1, Atg3, Atg12 expressions and LC3 cleavage in each cell line. Concomitantly, BiP, IRE1α and Calreticulin expressions were upregulated following 3 h curcumin exposure in MDA-MB-231 wt and GH+ cells. According to MTT cell viability assay, autocrine GH-mediated curcumin resistance was overcome by bafilomycin and curcumin co-treatment in MDA-MB-231 and T47D GH+ cells. Moreover, curcumin and bafilomycin co-treatment induced cell cycle arrest at G1 phase in MDA-MB-231 GH+ cells, G2/M arrest in T47D GH+ breast cancer cells. In conclusion, autocrine GH-triggered curcumin resistance was overcome by autophagy inhibition condition by bafilomycin treatment in a dose-dependent manner in MDA-MB-231 and T47D GH+ breast cancer cells. Full article

The aim of this project was the determination of the effect of bee bread supplement in diets of obese rats on interleukin 6 immunreactivity. This study has been shown that bee bread against obesity with a high-fat diet reduces IL-6 expression in the stomach. Full article

Pterygium is a benign fibrovascular proliferation that develops from the conjunctiva and invades the cornea. The etiology of this disorder remains unclear. Current treatment of pterygium is surgical. The postoperative recurrence rate of pterygium is reported be high. To the best of our knowledge, these results are the first to demonstrate the contribution of proteins expressions of the small GTPase Rho proteins in patients with pterygium. Our data showed that leukocyte RhoA, RhoB, RhoD, and RhoE protein expressions were markedly elevated in primary pterygium. However, no significant modifications were noted in pterygial tissues. Full article

Pterygium is one of the most common ocular surface diseases, and characterized by inflammatory infiltrates, proliferation, fibrosis, angiogenesis, and extracellular matrix breakdown. We investigated the association of polymorphisms in the RHO genes RHOA, RHOB, RHOC, RHOD, and RND3 (RHOE). The results of this study demonstrate for the first time the association of RHO genes with the pterygium. We displayed that the RHO gene polymorphisms were significantly associated with pterygium in the Turkish population. Full article

Nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly used as anti-inflammatory and analgesic agents. This family of drugs suppresses prostaglandin synthesis through inhibition of cyclooxygenase (COX) enzymes. Recent studies showed that anti-carcinogenic effects of these drugs are especially mediated by COX-2 enzyme. Etodolac is a COX-2 inhibitor and though not perfectly selective, it exhibits “preferential selectivity” for COX-2. In this study, the anti-proliferative and apoptotic effects of etodolac and its hydrazone or triazole derivatives (SGK 206 and SGK 242, respectively), were investigated on prostate cancer cell line PC-3 and human colorectal carcinoma cell line HT-29. Our data showed that SGK 206 and SGK 242 were more effective in the inhibition of proliferation and induction of apoptosis compared to etodolac in both cell lines. Full article

Lung cancer is one of the most diagnosed cancers worldwide and the development of anticancer agents for the treatment of lung cancer is an important task for researchers. For this purpose, herein pyrazoline-based compounds 1–18 were investigated for their cytotoxic effects on A549 human lung adenocarcinoma and CCD-19Lu human lung fibroblast cell lines using MTT assay. According to the results, 1-(4-methoxyphenyl)-3-(2-furanyl)-5-(4-methylphenyl)-2-pyrazoline (7) was identified as the most promising anticancer agent due to its selective inhibitory effect on A549 cell line with an IC₅₀ value of 138.63 µg/mL when compared with cyclophosphamide (IC₅₀ = 309.97 µg/mL). Full article

The present study investigates the total phenol amount, radical scavenging and anticancer activity of methanolic extract of Phlomis brevibracteata Turrill (PBT) leaves; an endemic plant of North Cyprus. The antiradical activity and cytotoxicity of 70% methanolic extract of PBT leaves were analyzed by the implementation of DPPH, ABTS radical scavenging activity tests and MTT assay, respectively. Our results showed significant antiradical scavenging activity in the total phenolic content of the PBT leaves. MTT assay proved potent cytotoxic activity against SK-HEP-1 cell line not only for 70% methanolic extract of PBT, but also for gallic acid and caffeic acid constituents of the total phenolic extract. Full article

Acute lymphoblastic leukemia (ALL) is the most common cancer type observed in childhood. Bone marrow (BM) microenvironment has significant role both at the beginning and the progress of leukemic period. Mesenchymal stem cells (MSCs) that make up microenvironment, regulate growth factors, cytokines and induction of survival of ALL cells by generating intracellular signals and establishing drug resistance. p97/VCP is a type of protein and it is responsible for also intracellular ubiquitin proteasome pathway, endoplasmic-reticulum-associated protein degradation, cell cycle, apoptosis and autophagy. It is shown that p97/VCP is overexpressed in the most of the cancer types including the ALL. p97/VCP inhibitors serve as therapeutic agents. Amongst p97/VCP inhibitors, DBeQ (dibenzylquinazoline-2,4-diamine) provide to undergo apoptosis and prevents cell proliferation. In this study, MSCs obtained from ALL patients and healthy donors were isolated by density-gradient method and then these cells were cultured. Cells were incubated with DBeQ. Cell apoptosis was determined by Annexin V/PI method (in flow cytometry). According to obtained data, availability of p97/VCP inhibitor which was recommended to be used as therapeutic agent in the treatment of other cancer types, in ALL treatment will be evaluated and this study will provide a basis for future studies. Full article

Human SH-SY5Y cell line has been used as an in vitro model in neuroscience research. However, many researchers emphasized that there are many differences between differentiated and undifferentiated characteristics of SH-SY5Y. Although Retinoic Acid (RA) generally have been used for differentiation of SH-SY5Y cells, which protocol provides better differentiation have not been cleared yet. Therefore, we compared RA and the other mediums in different treatment periods for obtaining better differentiated, carrying a neuron-like phenotype (N-type) characteristics, of SH-SY5Y human cells. The cells were pre-treated with different mediums for different treatment periods and compared with both for each other and the control group. When we pre-treated the cells with RA for 5-day and followed by the Mix medium (Neurobasal, B27, db-cAMP, KCI) and BDNF, neurite length and MAP2 expression of the cells were found significantly higher than the control group. In conclusion, we showed that more than one agent (RA) is clearly necessary to reach better differentiation of SH-SY5Y cells. Full article

OGG1 (8-oxo-G DNA glycosylase 1) is a BER (base excision repair) enzyme responsible for the repair of 8-oxo-G base damage related to oxidative stress through glycosylation. Determination of the interaction of DNA repair enzymes with other proteins that contributes to repairing base damages in the cell, is highly relevant in cancer treatment strategies and therapeutic targeting studies. In a previous study, it is shown that OGG1 associated with the homeobox protein NKX3.1 and androgen receptor (AR) in the 8-OHdG-repair complex in prostate cell line LNCaP, when treated with etoposide. Additionally, it is still uncertain that whether S326C, a polymorphic variant of OGG1 formed by a single amino acid change, might harbor functional loss, which might cause a deficiency in repair activity augmenting the susceptibility to (PCa). Therefore, in our study, we investigated the interaction of OGG1 and its polymorphic variant S326C with prostate specific proteins in LNCaP cells under oxidative conditions. Thus, we confirmed the previously determined interactions, however oxidative induction via menadione treatment impairs these interactions. Full article

Glioblastoma is the most common high-grade brain tumor in adults, which has a rapidly developing radiotherapy/chemotherapy resistance and high recurrence rate. The aim of our study is to determine the effects of bortezomib that is a quite important proteasome inhibitor on glioblastoma tumors in the 3D environment. In this study, we have performed bortezomib and temozolomide drug treatments and radiation applications to U251 cell line in 2D and 3D cultures with PCL/gelatin/graphene nanofibrous scaffolds. The best result was reached in the combination of drugs with radiation therapy that has the most decreased viability in the experiment group. Full article

Prostate cancer cells need androgens to grow and maintain like normal prostate cells, both utilize that Androgen Receptor (AR) function. Androgen receptor (AR) is expressed throughout the prostate cancer progression plays a critical role as a transcription factor in castration-dependent stages of disease. AR also interacts to many cellular proteins, including p53, to regulate apoptosis. Further, as the stabilization of p53 protein triggers apoptosis, p53 interacting small molecules such as Nutlin3a, are interpreted as cancer therapeutics. In this study, to find out how Nutlin3a-mediated p53 stabilization effect on AR signaling. Here, we investigated the dynamics of p53 binding to transcriptional targets of AR, and further investigated the variations of AR intracellular localization as well as transactivation in the presence of Nutlin3a. To do this, the changes in AR transactivation were investigated via luciferase reporter assay, which was performed by treating LNCaPs with different doses of Nutlin3a and resulted that transactivation was suppressed by Nutlin3a in a dose dependent manner. AR transactivation and sub-cellular localization were also studied by immunofluorescence assay and found that cytoplasmic-nuclear fractionation-coupled western blot analysis showed that Nutlin3a inhibits AR phosphorylation and nuclear translocation regardless of androgens. Full article

Increased neopterin levels reflect the activation of the cellular immune system which is of importance in the pathogenesis and progression of various diseases such as cancer. However, the effects of neopterin on the biological activity of Hepatocellular carcinoma cells are not yet illuminated. In this study we aim to find the effects of neopterin on viability/cytotxicity and motility of five different HCC cell lines with MTT, SRB and wound healing assays. According to our results, HCC cell lines can be grouped into two categories based on their proliferative and motility response to neopterin, as sensitive and resistant. This data provides us with variable results concerning the proliferation and motility of the cell lines under the effect of neopterin. The underlying molecular mechanisms will be examined in further studies, with a focus on the effect of neopterin on the signalling pathways. Overall, neopterin might have an important role in the hepatocarcinogenesis. Full article

Neuroblastomas are solid tumors and mostly seen in the adrenal medulla and sympathetic ganglia. It is known that neuroblastoma cell proliferation is inhibited by cisplatin and vincristine. The aim of this study was to investigate the effect of oxytocin on cell viability in human neuroblastoma SH-SY5Y cell line and primary cerebral cortex cell culture exposed to cisplatin and vincristine. In this direction, SH-SY5Y cell line and cortex neurons were obtained from the medical pharmacology department, Ataturk University. Both cells were grown in the appropriate cell culture media. Cisplatin (5, 10, 15 μg), vincristine (0.5, 1 and 2 ng) and oxytocin (1 μM) were applied to SH-SY5Y cell line and primary cortex cell culture for 24 h. MTT and TAC-TOS tests were performed 24 h after the application. As a result of the MTT assay, the combination of cisplatin and vincristine reduced cell viability in both cultures approximately 25% and 22%, respectively, compared to the control group. It appears that oxytocin increases neuroblastoma and cortex neuron viability, 112% and 95%, respectively. In this relation, we need to investigate why oxytocin increases cell viability and what are the possible implications in women in lactation stage. Full article

The ubiquitin proteasomal system and autophagic pathway are two main protein degradation systems in eukaryotic cells. Inhibition of the proteasomal system with proteasome inhibitors for cancer treatment can cause neurotoxic side effects. In this study, we investigated neurotoxic side effects of bortezomib (BTZ) and carfilzomib (CFZ) in a human neuronal cell model. Inhibition of proteasome with BTZ upregulated autophagy receptor protein p62 level. BTZ caused reduced mitochondrial mass per cell in a greater extent than CFZ. BTZ caused more clustering of mitochondria than CFZ. In conclusion, mitochondrial toxicity and autophagic upregulation with BTZ may be the reason for more severe neurotoxic profile than CFZ. Full article

Undesirable side effects that result from the random use of herbicides in developing countries are widespread. Imazamox is a widely used herbicide and has toxic effect on humans. Momordica charantia has been reported to possess many benefits and medicinal properties. Pycnogenol (PYC) is a natural plant extract from the bark of Pinus pinaster Aiton and has potent antioxidant activities. The aim of this study was to evaluate the protective effects of pycnogenol and momardica on induced imazamox toxicity effects on L929 fibroblast cell line. L929 fibroblast cells were cultured in the appropriate culture medium. Toxic concentration of imazamox 250 μM, were administered 30 min prior to momordica and pycnogenol (10−1–10−5 concentration) on L929 fibroblast cell line for 24 h. The cell viability assay was determined by using MTT test. TAC-TOS analysis were used to evaluate antioxidant and oxidant status. According to our study pycnogenol high dose showned protective effect whereas momordica low dose showned protective effect p < 0.05. In current study pycnogenol increased TAC capacity in high dose but in lower dose pycnogenol did not show any promise (p < 0.05). Momordica results showed correlation with MTT result. According to this analyse momordica only in low dose increased anti oxidant capacity and in addition, only in low dose TOS level were decreased (p > 0.05). In conclusion momordica and pycnogenol showed promise to reduced imazamox toxicity. Full article

Technological devices such as mobile phones, wi-fi and bluetooth applications used widely in daily life emit low-dose electromagnetic fields (EMFs). EMFs influence metabolic processes in the body and exert various harmful biological effects on cells. This study aims to evaluate acute and chronic effects of 900 MHz EMF exposure on the microscopic structure of rat liver. Twenty-four adult Wistar albino rats were randomly divided into sham-control (Group I), acutely EMF-exposed (Group II) and chronically EMF-exposed (Group III) groups (n = 8 in each). The Rats of Group II were exposed to 900 MHz EMF using a microwave test transmitter during 24 h continuously just for one day. Group III-rats were exposed to 900 MHz EMF for 60 minutes a day for 30 days. Sham-controls were not applied EMF. After the rats were sacrificed under the anesthesia, their livers were removed and processed for light microscopic evaluation. Liver sections stained with histochemical dyes (H&E, PAS and Masson’s trichrome) displayed many histopathological alterations in both of the EMF-exposed groups, including foci of necrosis, inflammation, excessive vacuolar degenerations and apoptosis in hepatocytes, apparent vascular expansions and haemorrhage. Additionally, mononuclear cell infiltrations, biliary hyperplasia, fibrosis in periportal and centrilobular areas and decreased Kupffer cell population were determined in the chronic EMF exposure group. In contrast, the amount of the Kupffer cells were much more in the acute exposure group. Our findings suggested that both acute and chronic exposure to 900 MHz EMFs can lead to hepatic injury in rats. Full article

Cervical cancer is one of the frequent types of cancer seen in females. It has been suggested that natural compounds can be used effectively for cancer treatment. Apoptosis and autophagy related cell death play important roles in suppression of tumorigenesis. Apigenin and curcumin are natural products isolated from plant extracts known to have antitumoral, antibacterial and antiviral effects. Varying doses of curcumin and apigenin were applied to HeLa cancer cell lines. The expression of the genes related to apoptosis and/or autophagy related cell death were measured using qRT-PCR and cell viability was measured using MTT assay. Our results showed that curcumin and apigenin are effective on apoptosis and autophagy related cell death in HeLa cells. We suggested that these natural products seem to be a new promising therapeutic approach in cancer. Full article

Cyclophosphamide (CTX), also known as cytophosphane among other, is a medication used as chemotherapy and to suppress the immune system. The PI3K/AKT/mTOR pathway is involved in the regulation of diverse cellular functions, including cell growth, protein synthesis, cell cycle regulation, glucose metabolism, and motility. In our study eight weeks old C57BL/6 female mice were divided into 3 groups as control (C), sham (S) and experimental group. The experimental group has been established with CTX treatment. No treatment was applied to the C group. The S group were given an equal amount of saline. CTX was administered intraperitoneally one every 2 days for 3 weeks; the first dose was 70 mg/kg, the ongoing doses were 30 mg/kg. At the end of 3 weeks mice were sacrificed and kidneys were taken for investigation. In order to show the effect of cyclophosphamide in kidney tissue, the tissues were stained via indirect immunohistochemistry with PI3K, AKT and mTOR primary antibodies. In our study, PI3K, AKT and mTOR expression levels were found to be significantly decreased in CTX-mediated mechanisms indicating that the mechanisms of CTX might involve in the inhibition of PI3K/AKT/mTOR signaling pathway. Full article

Lung cancer, among all cancer types around the world, is listed as the major cause of death with high mortality rate. ERG transcription factor has an important role in the Epithelial Mesenchymal Transition (EMT), which is one of the most important mechanisms in lung cancer progression. It is known that a small molecule inhibitor YK-4-279 is a potential inhibitor of ERG expression in prostate cancer. In this study, the possible synergistic effect of YK-4-279 and another anticancer drug, Paclitaxel is analysed in non-small cell lung cancer (NSCLC) cell line, A549. Full article

Lung cancer is listed as the most fatal type of cancer around the world. ERG, a transcription factor, is an ETS family member protein. It is involved in Epithelial Mesenchymal Transition (EMT), metastasis and DNA repair defects. Etoposide is a chemotherapeutic agent used in treatment of a variety of different cancer types, including lung cancer. Moreover, Etoposide has been tested alone and in combination with other drugs in order to inhibit DNA synthesis and exhibit antitumor activity. The purpose of this study is to examine the effect of Etoposide treatment on ERG transfected non-small cell lung cancer (NSCLC) cell line, H358. Full article

Genetic alterations affect carcinogenesis, and recent studies demonstrated that long non-coding RNAs (lncRNAs) have critical roles during this process. Hox transcript antisense intergenic RNA (HOTAIR) is a lncRNA molecule that affects proliferation, survival, migration, genomic stability, and drug resistance of cancer cells. The purpose of this study is to examine the possible effects of Gemcitabine treatment on HOTAIR expression level in non-small cell lung cancer (NSCLC) cell lines, H1944 and H596. Full article

Neuroblastoma is an embryonic tumor originating from the neural crest. It accounts for 8–10% of all childhood cancers. Although Cisplatin is used in neuroblastoma treatment, it has many side effects, such as ototoxicity, nephrotoxicity, and neurotoxicity. One herbal agent that has attracted attention in recent years is oleuropein (OLE), the active component of olive leaf. This component belongs to the polyphenol group and it has antioxidant, anti-microbial, anti-inflammatory, anti-hypertensive and anti-carcinogenic effects. It has beneficial effects against neurodegeneration in both culture cells and model organisms. Oleuropein has been shown to be increased apoptosis in SH-SY5Y neuroblastoma cell line in one study. Cisplatin (cis-diaminedichloroplatinum II (CDDP) is a widely used agent for the treatment of many different human cancers in childhood and adults with antimitotic and antineoplastic properties. CDDP is the most effective chemotherapeutic agent in specially treatment of neuroblastoma. Purpose of this study was to determine whether oleuropein and CDDP have possible anti-proliferative activity in different types of human neuroblastoma cells as representing different clinical features (bone marrow metastatic LAN-5 cells and treated with chemotherapy and beam therapy CHP-134 cells representing late relapse) investigated. Human bone marrow metastatic LAN-5 and treated with chemotherapy and beam therapy CHP-134 neuroblastoma cells representing late relapse were used in this study. The effects of OLE and CDDP on LAN-5 and CHP-134 neuroblastoma cell proliferation and apoptotic cell death was investigated using WST-1 cell proliferation and Annexin-V/PI flow cytometric assays. Oleuropein and CDDP have been shown to inhibit proliferation of LAN-5 and CHP-134 neuroblastoma cells. In further studies, it is planned to investigate different cell death mechanisms by using combination of oleuropein and cisplatin in different kind of human neuroblastoma cells. Full article

Bone marrow mesenchymal stem cells (BM-MSCs) are used to repair hypoxic or ischemic tissue. After hypoxic the level of ATP is decreases, cellular functions do not continue and apoptosis or necrosis occur. Apoptosis is a progress of programmed cell death that occurs in normal or pathological conditions. In this study, we were investigated the hypoxic effect on apoptosis in mesenchymal stem cell. Bone marrow-derived stem cells were cultured in hypoxic (1% or 3%) or normoxic conditions 24, 96 well plates for 36 h. Cell viability was shown by MTT assay on 36 h. After fixation of cells with 4% paraformaldehyde, distributions of caspase-3, Bcl-2 and Bax with indirect immunoperoxidase technique, apoptotic cells with TUNEL assay were investigated. All staining results were evaluated using H-score analyses method with ANOVA, statistically. As a result, hypoxic condition was toxic for human mesenchymal stem cells and the number of death cell was higher in that than normoxic condition. Full article

Among chemotherapeutic agents, cisplatin and the other platinum-based drugs have occupied for 35 years an enviable position. The limitations of platinum-based drugs, dose dependent side effects and development of drug resistance mechanisms, have boosted the research for finding other metal-based drugs. Among metals, ruthenium is probably the one showing the greatest promises. Ruthenium (Ru) appears to be less toxic than platinum and several biological studies have indicated that ruthenium complexes possess diverse modes of action. The redox chemistry of ruthenium is rich and compatible with biological media, and the overall toxicity of ruthenium is lower than platinum, thus allowing higher doses of treatment. In this study we aimed that, analyses of different type of ruthenium complexes in cancer cell lines. Six Ru complexes were determined by elemental analysis, FTIR, NMR, UV-visible spectroscopy, electron density on the metal was measured by cyclic voltammetry. After that, the cellular properties of this complexes were analyses on PC-3, HT-29, Du-145 and Vero cell lines. DNA damage was analyzed H2AX staining, apoptotic cell analyses were performed flow cytometry and western blotting. After 48 h incubation of Ru complexes three of them more effective for cell lines. Especially Ru3 was more effective in cancer cell lines. Apoptotic pathway was triggered after Ru complexes incubation in PC-3, Du-145 and Ht-29 cancer cell lines. Our study suggest that Ru complexes may be used for cancer cell cytotoxicity as a drugs in patients. Full article