A Fungicide, Fludioxonil, Formed the Polyploid Giant Cancer Cells and Induced Metastasis and Stemness in MDA-MB-231 Triple-Negative Breast Cancer Cells
Abstract
:1. Introduction
2. Results
2.1. Effects of Fludioxonil on the Cell Viability of Breast Cancer Cell Lines
2.2. Transformation to Polyploid Giant Cancer Cells
2.3. Cell Migration and Resistance to Anticancer Drugs in PGCCs Formed by Exposure to Fludioxonil
2.4. Effects of Fludioxonil on Change to Cell Pathways
2.5. Characteristics of the Daughter Cells Formed by Exposure to Fludioxonil
3. Materials and Methods
3.1. Chemicals
3.2. Cell Media, Culture, and Seeding
Cell Lines | ER | PR | HER2 | BRCA1 Mutation | Subtype | Tumor Type | p53 Gene |
---|---|---|---|---|---|---|---|
MDA-MB-231 | − | − | − | WT | TNB | AC | Mut |
T47D | + | + | − | WT | LA | IDC | Mut |
MCF-7 | + | + | − | WT | LA | IDC | WT |
3.3. WST Assay for Identification of Cell Viability
3.4. Morphology Staining
3.5. Live-Cell Image System for Identification of Cell Division
3.6. Cell Cycle Analysis Using the FACS
3.7. Scratch Assay to Evaluate Cell Migration
3.8. Evaluation of Chemo-Resistance
3.9. mRNA Sequencing and Analyses
3.10. Quantitative Reverse Transcription Polymerase Chain Reaction (qPCR)
3.11. Western Blot Analysis
3.12. Acquisition of Daughter Cells
3.13. Statistical Analysis
4. Discussion
Limitations of the Study
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Davoli, T.; de Lange, T. The causes and consequences of polyploidy in normal development and cancer. Annu. Rev. Cell Dev. Biol. 2011, 27, 585–610. [Google Scholar] [CrossRef] [PubMed]
- Liu, Z.; Yue, S.; Chen, X.; Kubin, T.; Braun, T. Regulation of Cardiomyocyte Polyploidy and Multinucleation by CyclinG1. Circ. Res. 2010, 106, 1498–1506. [Google Scholar] [CrossRef]
- Song, Y.; Zhao, Y.; Deng, Z.; Zhao, R.; Huang, Q. Stress-Induced Polyploid Giant Cancer Cells: Unique Way of Formation and Non-Negligible Characteristics. Front. Oncol. 2021, 11, 724781. [Google Scholar] [CrossRef]
- Vasan, N.; Toska, E.; Scaltriti, M. Overview of the relevance of PI3K pathway in HR-positive breast cancer. Ann. Oncol. 2019, 30 (Suppl. 10), x3–x11. [Google Scholar] [CrossRef]
- Erenpreisa, J.; Ivanov, A.; Wheatley, S.P.; Kosmacek, E.A.; Ianzini, F.; Anisimov, A.P.; Mackey, M.; Davis, P.J.; Plakhins, G.; Illidge, T.M. Endopolyploidy in irradiated p53-deficient tumour cell lines: Persistence of cell division activity in giant cells expressing Aurora-B kinase. Cell Biol. Int. 2008, 32, 1044–1056. [Google Scholar] [CrossRef]
- Zhang, S.; Mercado-Uribe, I.; Xing, Z.; Sun, B.; Kuang, J.; Liu, J. Generation of cancer stem-like cells through the formation of polyploid giant cancer cells. Oncogene 2014, 33, 116–128. [Google Scholar] [CrossRef]
- Saini, G.; Joshi, S.; Garlapati, C.; Li, H.; Kong, J.; Krishnamurthy, J.; Reid, M.D.; Aneja, R. Polyploid giant cancer cell characterization: New frontiers in predicting response to chemotherapy in breast cancer. Semin. Cancer Biol. 2022, 81, 220–231. [Google Scholar] [CrossRef] [PubMed]
- Chen, J.; Niu, N.; Zhang, J.; Qi, L.; Shen, W.; Donkena, K.V.; Feng, Z.; Liu, J. Polyploid Giant Cancer Cells (PGCCs): The Evil Roots of Cancer. Curr. Cancer Drug Targets 2019, 19, 360–367. [Google Scholar] [CrossRef]
- White-Gilbertson, S.; Voelkel-Johnson, C. Giants and monsters: Unexpected characters in the story of cancer recurrence. Adv. Cancer Res. 2020, 148, 201–232. [Google Scholar] [CrossRef]
- Steeg, P.S. Targeting metastasis. Nat. Rev. Cancer 2016, 16, 201–218. [Google Scholar] [CrossRef] [PubMed]
- Ashrafizadeh, M.; Hushmandi, K.; Hashemi, M.; Akbari, M.E.; Kubatka, P.; Raei, M.; Koklesova, L.; Shahinozzaman, M.; Mohammadinejad, R.; Najafi, M.; et al. Role of microRNA/Epithelial-to-Mesenchymal Transition Axis in the Metastasis of Bladder Cancer. Biomolecules 2020, 10, 1159. [Google Scholar] [CrossRef]
- Pijuan, J.; Barcelo, C.; Moreno, D.F.; Maiques, O.; Siso, P.; Marti, R.M.; Macia, A.; Panosa, A. In vitro Cell Migration, Invasion, and Adhesion Assays: From Cell Imaging to Data Analysis. Front. Cell Dev. Biol. 2019, 7, 107. [Google Scholar] [CrossRef]
- Diepenbruck, M.; Christofori, G. Epithelial-mesenchymal transition (EMT) and metastasis: Yes, no, maybe? Curr. Opin. Cell Biol. 2016, 43, 7–13. [Google Scholar] [CrossRef] [PubMed]
- Hashemi, M.; Arani, H.Z.; Orouei, S.; Fallah, S.; Ghorbani, A.; Khaledabadi, M.; Kakavand, A.; Tavakolpournegari, A.; Saebfar, H.; Heidari, H.; et al. EMT mechanism in breast cancer metastasis and drug resistance: Revisiting molecular interactions and biological functions. Biomed. Pharmacother. 2022, 155, 113774. [Google Scholar] [CrossRef] [PubMed]
- Campbell, L.L.; Polyak, K. Breast tumor heterogeneity: Cancer stem cells or clonal evolution? Cell Cycle 2007, 6, 2332–2338. [Google Scholar] [CrossRef] [PubMed]
- Clarke, M.F.; Dick, J.E.; Dirks, P.B.; Eaves, C.J.; Jamieson, C.H.; Jones, D.L.; Visvader, J.; Weissman, I.L.; Wahl, G.M. Cancer stem cells--perspectives on current status and future directions: AACR Workshop on cancer stem cells. Cancer Res. 2006, 66, 9339–9344. [Google Scholar] [CrossRef]
- Peiris-Pages, M.; Martinez-Outschoorn, U.E.; Pestell, R.G.; Sotgia, F.; Lisanti, M.P. Cancer stem cell metabolism. Breast Cancer Res. 2016, 18, 55. [Google Scholar] [CrossRef]
- Mauro-Lizcano, M.; Sotgia, F.; Lisanti, M.P. SOX2-high cancer cells exhibit an aggressive phenotype, with increases in stemness, proliferation and invasion, as well as higher metabolic activity and ATP production. Aging 2022, 14, 9877–9889. [Google Scholar] [CrossRef]
- Go, R.E.; Kim, C.W.; Jeon, S.Y.; Byun, Y.S.; Jeung, E.B.; Nam, K.H.; Choi, K.C. Fludioxonil induced the cancer growth and metastasis via altering epithelial-mesenchymal transition via an estrogen receptor-dependent pathway in cellular and xenografted breast cancer models. Environ. Toxicol. 2017, 32, 1439–1454. [Google Scholar] [CrossRef]
- Medjakovic, S.; Zoechling, A.; Gerster, P.; Ivanova, M.M.; Teng, Y.; Klinge, C.M.; Schildberger, B.; Gartner, M.; Jungbauer, A. Effect of nonpersistent pesticides on estrogen receptor, androgen receptor, and aryl hydrocarbon receptor. Environ. Toxicol. 2014, 29, 1201–1216. [Google Scholar] [CrossRef]
- Teng, Y.; Manavalan, T.T.; Hu, C.; Medjakovic, S.; Jungbauer, A.; Klinge, C.M. Endocrine disruptors fludioxonil and fenhexamid stimulate miR-21 expression in breast cancer cells. Toxicol. Sci. 2013, 131, 71–83. [Google Scholar] [CrossRef]
- Kugathas, S.; Audouze, K.; Ermler, S.; Orton, F.; Rosivatz, E.; Scholze, M.; Kortenkamp, A. Effects of Common Pesticides on Prostaglandin D2 (PGD2) Inhibition in SC5 Mouse Sertoli Cells, Evidence of Binding at the COX-2 Active Site, and Implications for Endocrine Disruption. Environ. Health Perspect. 2016, 124, 452–459. [Google Scholar] [CrossRef]
- Graiet, I.; Hamdi, H.; Abid-Essefi, S.; Eyer, J. Fludioxonil, a phenylpyrrol pesticide, induces Cytoskeleton disruption, DNA damage and apoptosis via oxidative stress on rat glioma cells. Food Chem. Toxicol. 2022, 170, 113464. [Google Scholar] [CrossRef]
- Lee, G.H.; Hwang, K.A.; Choi, K.C. Effects of Fludioxonil on the Cell Growth and Apoptosis in T and B Lymphocytes. Biomolecules 2019, 9, 500. [Google Scholar] [CrossRef] [PubMed]
- Karadag, H.; Ozhan, F. Effect of cyprodinil and fludioxonil pesticides on bovine liver catalase activity. Biotechnol. Biotechnol. Equip. 2015, 29, 40–44. [Google Scholar] [CrossRef] [PubMed]
- Lee, S.M.; Ko, E.B.; Go, R.E.; Lee, H.K.; Choi, K.C. Effect of the phenylpyrrole fungicide fludioxonil on cell proliferation and cardiac differentiation in mouse embryonic stem cells. Reprod. Toxicol. 2021, 104, 76–84. [Google Scholar] [CrossRef]
- Graillot, V.; Takakura, N.; Hegarat, L.L.; Fessard, V.; Audebert, M.; Cravedi, J.P. Genotoxicity of pesticide mixtures present in the diet of the French population. Environ. Mol. Mutagen. 2012, 53, 173–184. [Google Scholar] [CrossRef] [PubMed]
- Gstraunthaler, G.; Lindl, T. Zell-und Gewebekultur; Springer Spectrum: Berlin, Germany, 2013; pp. 69–97. [Google Scholar] [CrossRef]
- Wasielewski, M.; Elstrodt, F.; Klijn, J.G.; Berns, E.M.; Schutte, M. Thirteen new p53 gene mutants identified among 41 human breast cancer cell lines. Breast Cancer Res. Treat. 2006, 99, 97–101. [Google Scholar] [CrossRef]
- Dai, X.; Cheng, H.; Bai, Z.; Li, J. Breast Cancer Cell Line Classification and Its Relevance with Breast Tumor Subtyping. J. Cancer 2017, 8, 3131–3141. [Google Scholar] [CrossRef]
- Kim, M.; Moon, A. A curcumin analog CA-5f inhibits urokinase-type plasminogen activator and invasive phenotype of triple-negative breast cancer cells. Toxicol. Res. 2022, 38, 19–26. [Google Scholar] [CrossRef]
- Pathak, V.M.; Verma, V.K.; Rawat, B.S.; Kaur, B.; Babu, N.; Sharma, A.; Dewali, S.; Yadav, M.; Kumari, R.; Singh, S.; et al. Current status of pesticide effects on environment, human health and it’s eco-friendly management as bioremediation: A comprehensive review. Front. Microbiol. 2022, 13, 962619. [Google Scholar] [CrossRef] [PubMed]
- U.S. Environmental Protection Agency (EPA). Exposure Assessment Tools by Chemical Classes—Pesticides. Available online: https://www.epa.gov/expobox/exposure-assessment-tools-chemical-classes-pesticides (accessed on 29 July 2024).
- Syafrudin, M.; Kristanti, R.A.; Yuniarto, A.; Hadibarata, T.; Rhee, J.; Al-Onazi, W.A.; Algarni, T.S.; Almarri, A.H.; Al-Mohaimeed, A.M. Pesticides in Drinking Water—A Review. Int. J. Environ. Res. Public. Health 2021, 18, 468. [Google Scholar] [CrossRef]
- Kalyabina, V.P.; Esimbekova, E.N.; Kopylova, K.V.; Kratasyuk, V.A. Pesticides: Formulants, distribution pathways and effects on human health—A review. Toxicol. Rep. 2021, 8, 1179–1192. [Google Scholar] [CrossRef] [PubMed]
- Beranger, R.; Hardy, E.M.; Binter, A.C.; Charles, M.A.; Zaros, C.; Appenzeller, B.M.R.; Chevrier, C. Multiple pesticides in mothers’ hair samples and children’s measurements at birth: Results from the French national birth cohort (ELFE). Int. J. Hyg. Environ. Health 2020, 223, 22–33. [Google Scholar] [CrossRef]
- Hainaut, P.; Hollstein, M. p53 and human cancer: The first ten thousand mutations. Adv. Cancer Res. 2000, 77, 81–137. [Google Scholar] [CrossRef]
- Walerych, D.; Napoli, M.; Collavin, L.; Del Sal, G. The rebel angel: Mutant p53 as the driving oncogene in breast cancer. Carcinogenesis 2012, 33, 2007–2017. [Google Scholar] [CrossRef]
- Oren, M.; Rotter, V. Mutant p53 gain-of-function in cancer. Cold Spring Harb. Perspect. Biol. 2010, 2, a001107. [Google Scholar] [CrossRef]
- Muller, P.A.; Vousden, K.H. p53 mutations in cancer. Nat. Cell Biol. 2013, 15, 2–8. [Google Scholar] [CrossRef] [PubMed]
- Asher, G.; Lotem, J.; Tsvetkov, P.; Reiss, V.; Sachs, L.; Shaul, Y. P53 hot-spot mutants are resistant to ubiquitin-independent degradation by increased binding to NAD(P)H:quinone oxidoreductase 1. Proc. Natl. Acad. Sci. USA 2003, 100, 15065–15070. [Google Scholar] [CrossRef]
- Hui, L.; Zheng, Y.; Yan, Y.; Bargonetti, J.; Foster, D.A. Mutant p53 in MDA-MB-231 breast cancer cells is stabilized by elevated phospholipase D activity and contributes to survival signals generated by phospholipase D. Oncogene 2006, 25, 7305–7310. [Google Scholar] [CrossRef]
- Zeng, J.; Hills, S.A.; Ozono, E.; Diffley, J.F.X. Cyclin E-induced replicative stress drives p53-dependent whole-genome duplication. Cell 2023, 186, 528–542.e514. [Google Scholar] [CrossRef] [PubMed]
- Davoli, T.; Denchi, E.L.; de Lange, T. Persistent telomere damage induces bypass of mitosis and tetraploidy. Cell 2010, 141, 81–93. [Google Scholar] [CrossRef]
- Narkar, A.; Johnson, B.A.; Bharne, P.; Zhu, J.; Padmanaban, V.; Biswas, D.; Fraser, A.; Iglesias, P.A.; Ewald, A.J.; Li, R. On the role of p53 in the cellular response to aneuploidy. Cell Rep. 2021, 34, 108892. [Google Scholar] [CrossRef]
- Bortner, D.M.; Rosenberg, M.P. Induction of mammary gland hyperplasia and carcinomas in transgenic mice expressing human cyclin E. Mol. Cell. Biol. 1997, 17, 453–459. [Google Scholar] [CrossRef]
- Parisi, T.; Beck, A.R.; Rougier, N.; McNeil, T.; Lucian, L.; Werb, Z.; Amati, B. Cyclins E1 and E2 are required for endoreplication in placental trophoblast giant cells. EMBO J. 2003, 22, 4794–4803. [Google Scholar] [CrossRef]
- Geng, Y.; Yu, Q.; Sicinska, E.; Das, M.; Schneider, J.E.; Bhattacharya, S.; Rideout, W.M.; Bronson, R.T.; Gardner, H.; Sicinski, P. Cyclin E ablation in the mouse. Cell 2003, 114, 431–443. [Google Scholar] [CrossRef] [PubMed]
- Gao, S.; Ma, J.J.; Lu, C. Prognostic value of cyclin E expression in breast cancer: A meta-analysis. Tumour Biol. 2013, 34, 3423–3430. [Google Scholar] [CrossRef]
- Teixeira, L.K.; Wang, X.; Li, Y.; Ekholm-Reed, S.; Wu, X.; Wang, P.; Reed, S.I. Cyclin E deregulation promotes loss of specific genomic regions. Curr. Biol. 2015, 25, 1327–1333. [Google Scholar] [CrossRef] [PubMed]
- Kim, C.W.; Go, R.E.; Choi, K.C. Treatment of BG-1 Ovarian Cancer Cells Expressing Estrogen Receptors with Lambda-cyhalothrin and Cypermethrin Caused a Partial Estrogenicity via an Estrogen Receptor-dependent Pathway. Toxicol. Res. 2015, 31, 331–337. [Google Scholar] [CrossRef]
- Etemadmoghadam, D.; Au-Yeung, G.; Wall, M.; Mitchell, C.; Kansara, M.; Loehrer, E.; Batzios, C.; George, J.; Ftouni, S.; Weir, B.A.; et al. Resistance to CDK2 inhibitors is associated with selection of polyploid cells in CCNE1-amplified ovarian cancer. Clin. Cancer Res. 2013, 19, 5960–5971. [Google Scholar] [CrossRef]
- Yeudall, W.A.; Vaughan, C.A.; Miyazaki, H.; Ramamoorthy, M.; Choi, M.Y.; Chapman, C.G.; Wang, H.; Black, E.; Bulysheva, A.A.; Deb, S.P.; et al. Gain-of-function mutant p53 upregulates CXC chemokines and enhances cell migration. Carcinogenesis 2012, 33, 442–451. [Google Scholar] [CrossRef]
- Solomon, H.; Buganim, Y.; Kogan-Sakin, I.; Pomeraniec, L.; Assia, Y.; Madar, S.; Goldstein, I.; Brosh, R.; Kalo, E.; Beatus, T.; et al. Various p53 mutant proteins differently regulate the Ras circuit to induce a cancer-related gene signature. J. Cell Sci. 2012, 125 Pt 13, 3144–3152. [Google Scholar] [CrossRef] [PubMed]
- Solomon, H.; Madar, S.; Rotter, V. Mutant p53 gain of function is interwoven into the hallmarks of cancer. J. Pathol. 2011, 225, 475–478. [Google Scholar] [CrossRef]
- Cooks, T.; Pateras, I.S.; Tarcic, O.; Solomon, H.; Schetter, A.J.; Wilder, S.; Lozano, G.; Pikarsky, E.; Forshew, T.; Rosenfeld, N.; et al. Mutant p53 prolongs NF-kappaB activation and promotes chronic inflammation and inflammation-associated colorectal cancer. Cancer Cell 2013, 23, 634–646. [Google Scholar] [CrossRef]
- Karin, M.; Greten, F.R. NF-kappaB: Linking inflammation and immunity to cancer development and progression. Nat. Rev. Immunol. 2005, 5, 749–759. [Google Scholar] [CrossRef]
- Taniguchi, K.; Karin, M. NF-kappaB, inflammation, immunity and cancer: Coming of age. Nat. Rev. Immunol. 2018, 18, 309–324. [Google Scholar] [CrossRef]
- Elinav, E.; Nowarski, R.; Thaiss, C.A.; Hu, B.; Jin, C.; Flavell, R.A. Inflammation-induced cancer: Crosstalk between tumours, immune cells and microorganisms. Nat. Rev. Cancer 2013, 13, 759–771. [Google Scholar] [CrossRef] [PubMed]
- Godwin, P.; Baird, A.M.; Heavey, S.; Barr, M.P.; O’Byrne, K.J.; Gately, K. Targeting nuclear factor-kappa B to overcome resistance to chemotherapy. Front. Oncol. 2013, 3, 120. [Google Scholar] [CrossRef]
- Pires, B.R.; Mencalha, A.L.; Ferreira, G.M.; de Souza, W.F.; Morgado-Diaz, J.A.; Maia, A.M.; Correa, S.; Abdelhay, E.S. NF-kappaB Is Involved in the Regulation of EMT Genes in Breast Cancer Cells. PLoS ONE 2017, 12, e0169622. [Google Scholar] [CrossRef] [PubMed]
- Shigdar, S.; Li, Y.; Bhattacharya, S.; O’Connor, M.; Pu, C.; Lin, J.; Wang, T.; Xiang, D.; Kong, L.; Wei, M.Q.; et al. Inflammation and cancer stem cells. Cancer Lett. 2014, 345, 271–278. [Google Scholar] [CrossRef]
- Shimizu, T.; Marusawa, H.; Endo, Y.; Chiba, T. Inflammation-mediated genomic instability: Roles of activation-induced cytidine deaminase in carcinogenesis. Cancer Sci. 2012, 103, 1201–1206. [Google Scholar] [CrossRef]
- Lee, G.H.; Jin, S.W.; Kim, S.J.; Pham, T.H.; Choi, J.H.; Jeong, H.G. Tetrabromobisphenol A Induces MMP-9 Expression via NADPH Oxidase and the activation of ROS, MAPK, and Akt Pathways in Human Breast Cancer MCF-7 Cells. Toxicol. Res. 2019, 35, 93–101. [Google Scholar] [CrossRef] [PubMed]
- Weisz, L.; Damalas, A.; Liontos, M.; Karakaidos, P.; Fontemaggi, G.; Maor-Aloni, R.; Kalis, M.; Levrero, M.; Strano, S.; Gorgoulis, V.G.; et al. Mutant p53 enhances nuclear factor kappaB activation by tumor necrosis factor alpha in cancer cells. Cancer Res. 2007, 67, 2396–2401. [Google Scholar] [CrossRef]
- Scian, M.J.; Stagliano, K.E.; Anderson, M.A.; Hassan, S.; Bowman, M.; Miles, M.F.; Deb, S.P.; Deb, S. Tumor-derived p53 mutants induce NF-kappaB2 gene expression. Mol. Cell. Biol. 2005, 25, 10097–10110. [Google Scholar] [CrossRef]
- Schneider, G.; Henrich, A.; Greiner, G.; Wolf, V.; Lovas, A.; Wieczorek, M.; Wagner, T.; Reichardt, S.; von Werder, A.; Schmid, R.M.; et al. Cross talk between stimulated NF-kappaB and the tumor suppressor p53. Oncogene 2010, 29, 2795–2806. [Google Scholar] [CrossRef] [PubMed]
- Rivlin, N.; Brosh, R.; Oren, M.; Rotter, V. Mutations in the p53 Tumor Suppressor Gene: Important Milestones at the Various Steps of Tumorigenesis. Genes. Cancer 2011, 2, 466–474. [Google Scholar] [CrossRef]
- Brosh, R.; Rotter, V. When mutants gain new powers: News from the mutant p53 field. Nat. Rev. Cancer 2009, 9, 701–713. [Google Scholar] [CrossRef]
- Salmina, K.; Vainshelbaum, N.M.; Kreishmane, M.; Inashkina, I.; Cragg, M.S.; Pjanova, D.; Erenpreisa, J. The Role of Mitotic Slippage in Creating a “Female Pregnancy-like System” in a Single Polyploid Giant Cancer Cell. Int. J. Mol. Sci. 2023, 24, 3237. [Google Scholar] [CrossRef] [PubMed]
- Weihua, Z.; Lin, Q.; Ramoth, A.J.; Fan, D.; Fidler, I.J. Formation of solid tumors by a single multinucleated cancer cell. Cancer 2011, 117, 4092–4099. [Google Scholar] [CrossRef]
- Liu, J. The “life code”: A theory that unifies the human life cycle and the origin of human tumors. Semin. Cancer Biol. 2020, 60, 380–397. [Google Scholar] [CrossRef]
- Liu, J. The dualistic origin of human tumors. Semin. Cancer Biol. 2018, 53, 1–16. [Google Scholar] [CrossRef] [PubMed]
- Solari, F.; Domenget, C.; Gire, V.; Woods, C.; Lazarides, E.; Rousset, B.; Jurdic, P. Multinucleated cells can continuously generate mononucleated cells in the absence of mitosis: A study of cells of the avian osteoclast lineage. J. Cell Sci. 1995, 108 Pt 10, 3233–3241. [Google Scholar] [CrossRef] [PubMed]
- Zhang, D.; Yang, X.; Yang, Z.; Fei, F.; Li, S.; Qu, J.; Zhang, M.; Li, Y.; Zhang, X.; Zhang, S. Daughter Cells and Erythroid Cells Budding from PGCCs and Their Clinicopathological Significances in Colorectal Cancer. J. Cancer 2017, 8, 469–478. [Google Scholar] [CrossRef]
- Zhang, S.; Mercado-Uribe, I.; Liu, J. Generation of erythroid cells from fibroblasts and cancer cells in vitro and in vivo. Cancer Lett. 2013, 333, 205–212. [Google Scholar] [CrossRef]
- Zhang, S.; Mercado-Uribe, I.; Hanash, S.; Liu, J. iTRAQ-based proteomic analysis of polyploid giant cancer cells and budding progeny cells reveals several distinct pathways for ovarian cancer development. PLoS ONE 2013, 8, e80120. [Google Scholar] [CrossRef] [PubMed]
- Salmina, K.; Jankevics, E.; Huna, A.; Perminov, D.; Radovica, I.; Klymenko, T.; Ivanov, A.; Jascenko, E.; Scherthan, H.; Cragg, M.; et al. Up-regulation of the embryonic self-renewal network through reversible polyploidy in irradiated p53-mutant tumour cells. Exp. Cell Res. 2010, 316, 2099–2112. [Google Scholar] [CrossRef] [PubMed]
- Fei, F.; Liu, K.; Li, C.; Du, J.; Wei, Z.; Li, B.; Li, Y.; Zhang, Y.; Zhang, S. Molecular Mechanisms by Which S100A4 Regulates the Migration and Invasion of PGCCs with Their Daughter Cells in Human Colorectal Cancer. Front. Oncol. 2020, 10, 182. [Google Scholar] [CrossRef] [PubMed]
- Fei, F.; Zhang, D.; Yang, Z.; Wang, S.; Wang, X.; Wu, Z.; Wu, Q.; Zhang, S. The number of polyploid giant cancer cells and epithelial-mesenchymal transition-related proteins are associated with invasion and metastasis in human breast cancer. J. Exp. Clin. Cancer Res. 2015, 34, 158. [Google Scholar] [CrossRef]
- Wang, X.; Zheng, M.; Fei, F.; Li, C.; Du, J.; Liu, K.; Li, Y.; Zhang, S. EMT-related protein expression in polyploid giant cancer cells and their daughter cells with different passages after triptolide treatment. Med. Oncol. 2019, 36, 82. [Google Scholar] [CrossRef]
Media Composition | Cell Culture Media Cell Seeding Media | Phenol-Free Media |
---|---|---|
Medium | DMEM | DMEM |
Phenol red | Present | Absent |
Glucose concentration | High (4500 mg/L) | Low (1000 mg/L) |
FBS | 5% | 10% CD-FBS |
Penicillin/streptomycin | 2% | 2% |
HEPES | 2% (20 mM) | 2% (20 mM) |
L-glutamine | 4 mM | 1 mM |
Day | 1 | 2 | 3 | 5 | 6 | 7 | 9 | 10 |
---|---|---|---|---|---|---|---|---|
WST assay Staining Cell cycle analysis RNA extraction Protein extraction | Seeding (plate) | MC (p/f) | CT | MC | Exp | - | - | - |
Live-cell image | Seeding (plate) | MC (p/f) | CT and live start | MC | End | - | - | - |
Migration assay | Seeding (90 mm) | MC (p/f) | CT | MC | Tryp&S (plate) | MMC and scratch | Exp | - |
Anticancer drug resistance | Seeding (90 mm) | MC (p/f) | CT | MC | Tryp&S (plate) | ACD | - | Exp |
Genes | Primer Bank ID | Sequence (5′-3′) | Tm (°C) | Size (bp) | |
---|---|---|---|---|---|
TNF | 25952110c1 | F | CCTCTCTCTAATCAGCCCTCTG | 60.8 | 220 |
R | GAGGACCTGGGAGTAGATGAG | 60.2 | |||
CCL20 | 4759076a1 | F | TGCTGTACCAAGAGTTTGCTC | 60.2 | 220 |
R | CGCACACAGACAACTTTTTCTTT | 60.4 | |||
IL-1β | 27894305c1 | F | ATGATGGCTTATTACAGTGGCAA | 60.0 | 132 |
R | GTCGGAGATTCGTAGCTGGA | 60.8 | |||
MMP3 | 73808272c1 | F | AGTCTTCCAATCCTACTGTTGCT | 61.0 | 226 |
R | TCCCCGTCACCTCCAATCC | 63.0 | |||
CXCL3 | 4504157a1 | F | CGCCCAAACCGAAGTCATAG | 60.8 | 109 |
R | GCTCCCCTTGTTCAGTATCTTTT | 60.2 | |||
TRAF1 | 300193044c1 | F | TCCTGTGGAAGATCACCAATGT | 61.0 | 117 |
R | GCAGGCACAACTTGTAGCC | 61.3 | |||
IL-6 | 224831235c1 | F | ACTCACCTCTTCAGAACGAATTG | 60.2 | 149 |
R | CCATCTTTGGAAGGTTCAGGTTG | 61.3 | |||
PTGS2 | 223941909c1 | F | CTGGCGCTCAGCCATACAG | 62.8 | 94 |
R | CGCACTTATACTGGTCAAATCCC | 61.0 | |||
ICAM | 167466197c1 | F | ATGCCCAGACATCTGTGTCC | 61.6 | 112 |
R | GGGGTCTCTATGCCCAACAA | 61.3 | |||
ETV7 | 333470740c1 | F | CTGCTGTGGGATTACGTGTATC | 60.5 | 138 |
R | GTTCTTGTGATTTCCCCAGAGTC | 60.8 | |||
SLC12A7 | 123701899c1 | F | CTGGCGGGTCCTACTACATGA | 62.8 | 127 |
R | AAAATCTCGATGGTCCCCAAAAT | 60.2 | |||
S100P | 45827727c1 | F | AAGGATGCCGTGGATAAATTGC | 61.3 | 79 |
R | ACACGATGAACTCACTGAAGTC | 60.0 | |||
MMP13 | 296010793c1 | F | ACTGAGAGGCTCCGAGAAATG | 61.3 | 103 |
R | GAACCCCGCATCTTGGCTT | 62.7 | |||
18s-rRNA | 14165467c1 | F | GCGGCGGAAAATAGCCTTTG | 62.3 | 139 |
R | GATCACACGTTCCACCTCATC | 60.4 |
Protein | Manufacturer | Protein Size | Cat. No. | Dilution Ratio |
---|---|---|---|---|
Cyclin D1 | Biolegend | 36 | 681902 | 1:20 |
Cyclin E1 | Santacruz | 53 | sc-377100 | 1:20 |
Cyclin A1 | Bioss | 51 | bs-5739R | 1:10 |
Cyclin B1 | Biolegend | 48 | 647901 | 1:20 |
p53 | Santacruz | 53 | sc-126 | 1:10 |
E-cadherin | Biolegend | 100 | 324102 | 1:20 |
Vimentin | Thermo Fisher | 58 | MA5-11883 | 1:20 |
Nanog | Bioss | 34 | bs-0829R | 1:10 |
SOX2 | Biolegend | 35 | 656102 | 1:20 |
NF-κB p50(p105) | Biolegend | 50,100 | 616701 | 1:20 |
ERK1/2 | Cell signaling | 42,44 | 4695T | 1:10 |
p-ERK1/2 (Thr202/Tyr204) | Biolegend | 42,44 | 369501 | 1:10 |
GAPDH | Abcam | 40 | ab8245 | 1:100 |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Go, R.-E.; Seong, S.-M.; Choi, Y.; Choi, K.-C. A Fungicide, Fludioxonil, Formed the Polyploid Giant Cancer Cells and Induced Metastasis and Stemness in MDA-MB-231 Triple-Negative Breast Cancer Cells. Int. J. Mol. Sci. 2024, 25, 9024. https://doi.org/10.3390/ijms25169024
Go R-E, Seong S-M, Choi Y, Choi K-C. A Fungicide, Fludioxonil, Formed the Polyploid Giant Cancer Cells and Induced Metastasis and Stemness in MDA-MB-231 Triple-Negative Breast Cancer Cells. International Journal of Molecular Sciences. 2024; 25(16):9024. https://doi.org/10.3390/ijms25169024
Chicago/Turabian StyleGo, Ryeo-Eun, Su-Min Seong, Youngdong Choi, and Kyung-Chul Choi. 2024. "A Fungicide, Fludioxonil, Formed the Polyploid Giant Cancer Cells and Induced Metastasis and Stemness in MDA-MB-231 Triple-Negative Breast Cancer Cells" International Journal of Molecular Sciences 25, no. 16: 9024. https://doi.org/10.3390/ijms25169024