Endoplasmic Reticulum Stress Contributes to Gefitinib-Induced Apoptosis in Glioma
Abstract
:1. Introduction
2. Results
2.1. Gefitinib Induced ER Stress in H4 Cells
2.2. 4-Phenylbutyrate and IRE1 Silencing Alleviated Gefitinib-Induced Glioma Apoptosis in H4 Cells
2.3. BAPTA-AM and N-Acetyl-Cysteine (NAC) Alleviated Gefitinib-Induced Glioma Apoptosis in H4 Cells
2.4. Gefitinib Induced ER Stress and Apoptosis in U87 Cells
2.5. Gefitinib Caused Noxa Upregulation in Glioma Cells
3. Discussion
4. Materials and Methods
4.1. Cell Cultures
4.2. Cell Viability Assay
4.3. Caspase 3 Activity Assay
4.4. Measurement of ROS
4.5. Measurement of Cytosolic Ca2+
4.6. Western Blot
4.7. Small Interfering RNA (siRNA) Transfection
4.8. Statistical Analyses
Author Contributions
Funding
Conflicts of Interest
References
- Han, B.; Wang, H.; Zhang, J.; Tian, J. FNDC3B is associated with ER stress and poor prognosis in cervical cancer. Oncol. Lett. 2020, 19, 406–414. [Google Scholar] [CrossRef] [Green Version]
- Kwon, D.; Koh, J.; Kim, S.; Go, H.; Min, H.S.; Kim, Y.A.; Kim, D.K.; Jeon, Y.K.; Chung, D.H. Overexpression of endoplasmic reticulum stress-related proteins, XBP1s and GRP78, predicts poor prognosis in pulmonary adenocarcinoma. Lung Cancer 2018, 122, 131–137. [Google Scholar] [CrossRef]
- Kim, S.J.; Jegal, K.H.; Im, J.H.; Park, G.; Kim, S.; Jeong, H.G.; Cho, I.J.; Kang, K.W. Involvement of ER stress and reactive oxygen species generation in anti-cancer effect of CKD-516 for lung cancer. Cancer Chemother. Pharmacol. 2020, 85, 685–697. [Google Scholar] [CrossRef]
- Oakes, S.A. Endoplasmic reticulum stress signaling in cancer cells. Am. J. Pathol. 2020, 190, 934–946. [Google Scholar] [CrossRef] [Green Version]
- Stupp, R.; Taillibert, S.; Kanner, A.; Read, W.; Steinberg, D.; Lhermitte, B.; Toms, S.; Idbaih, A.; Ahluwalia, M.S.; Fink, K.; et al. Effect of tumor-treating fields plus maintenance temozolomide vs maintenance temozolomide alone on survival in patients with glioblastoma: A randomized clinical trial. JAMA 2017, 318, 2306–2316. [Google Scholar] [CrossRef] [Green Version]
- Hao, Z.; Guo, D. EGFR mutation: Novel prognostic factor associated with immune infiltration in lower-grade glioma; an exploratory study. BMC Cancer 2019, 19, 1184. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Muñoz-Hidalgo, L.; San-Miguel, T.; Megías, J.; Monleón, D.; Navarro, L.; Roldán, P.; Cerdá-Nicolás, M.; López-Ginés, C. Somatic copy number alterations are associated with EGFR amplification and shortened survival in patients with primary glioblastoma. Neoplasia 2020, 22, 10–21. [Google Scholar] [CrossRef] [PubMed]
- Liu, X.; Chen, X.; Shi, L.; Shan, Q.; Cao, Q.; Yue, C.; Li, H.; Li, S.; Wang, J.; Gao, S.; et al. The third-generation EGFR inhibitor AZD9291 overcomes primary resistance by continuously blocking ERK signaling in glioblastoma. J. Exp. Clin. Cancer Res. 2019, 38, 219. [Google Scholar] [CrossRef] [PubMed]
- Wu, S.; Gao, F.; Zheng, S.; Zhang, C.; Martinez-Ledesma, E.; Ezhilarasan, R.; Ding, J.; Li, X.; Feng, N.; Multani, A.; et al. EGFR amplification induces increased DNA damage response and renders selective sensitivity to Talazoparib (PARP inhibitor) in glioblastoma. Clin. Cancer Res. 2020, 26, 1395–1407. [Google Scholar] [CrossRef]
- Ye, C.; Pan, B.; Xu, H.; Zhao, Z.; Shen, J.; Lu, J.; Yu, R.; Liu, H. Co-delivery of GOLPH3 siRNA and gefitinib by cationic lipid-PLGA nanoparticles improves EGFR-targeted therapy for glioma. J. Mol. Med. 2019, 97, 1575–1588. [Google Scholar] [CrossRef]
- Prados, M.D.; Yung, W.K.; Wen, P.Y.; Junck, L.; Cloughesy, T.; Fink, K.; Chang, S.; Robins, H.I.; Dancey, J.; Kuhn, J. Phase-1 trial of gefitinib and temozolomide in patients with malignant glioma: A North American brain tumor consortium study. Cancer Chemother. Pharmacol. 2008, 61, 1059–1067. [Google Scholar] [CrossRef] [PubMed]
- Auf, G.; Jabouille, A.; Guérit, S.; Pineau, R.; Delugin, M.; Bouchecareilh, M.; Magnin, N.; Favereaux, A.; Maitre, M.; Gaiser, T.; et al. Inositol-requiring enzyme 1alpha is a key regulator of angiogenesis and invasion in malignant glioma. Proc. Natl. Acad. Sci. USA 2010, 107, 15553–15558. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Chen, Y.H.; Cimino, P.J.; Luo, J.; Dahiya, S.; Gutmann, D.H. ABCG1 maintains high-grade glioma survival in vitro and in vivo. Oncotarget 2016, 7, 23416–23424. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Peñaranda-Fajardo, N.M.; Meijer, C.; Liang, Y.; Dijkstra, B.M.; Aguirre-Gamboa, R.; den Dunnen, W.F.A.; Kruyt, F.A.E. ER stress and UPR activation in glioblastoma: Identification of a noncanonical PERK mechanism regulating GBM stem cells through SOX2 modulation. Cell Death Dis. 2019, 10, 690. [Google Scholar] [CrossRef] [PubMed]
- Chang, C.Y.; Li, J.R.; Wu, C.C.; Wang, J.D.; Liao, S.L.; Chen, W.Y.; Wang, W.Y.; Chen, C.J. Endoplasmic reticulum stress contributes to indomethacin-induced glioma apoptosis. Int. J. Mol. Sci. 2020, 21, 557. [Google Scholar] [CrossRef] [Green Version]
- Chang, C.Y.; Pan, P.H.; Li, J.R.; Ou, Y.C.; Wang, J.D.; Liao, S.L.; Chen, W.Y.; Wang, W.Y.; Chen, C.J. Aspirin induced glioma apoptosis through Noxa upregulation. Int. J. Mol. Sci. 2020, 21, 4219. [Google Scholar] [CrossRef]
- Wang, J.; Qi, Q.; Zhou, W.; Feng, Z.; Huang, B.; Chen, A.; Zhang, D.; Li, W.; Zhang, Q.; Jiang, Z.; et al. Inhibition of glioma growth by flavokawain B is mediated through endoplasmic reticulum stress induced autophagy. Autophagy 2018, 14, 2007–2022. [Google Scholar] [CrossRef] [Green Version]
- Ye, T.; Wei, L.; Shi, J.; Jiang, K.; Xu, H.; Hu, L.; Kong, L.; Zhang, Y.; Meng, S.; Piao, H. Sirtuin1 activator SRT2183 suppresses glioma cell growth involving activation of endoplasmic reticulum stress pathway. BMC Cancer 2019, 19, 706. [Google Scholar] [CrossRef]
- Zhang, Y.; Pusch, S.; Innes, J.; Sidlauskas, K.; Ellis, M.; Lau, J.; El-Hassan, T.; Aley, N.; Launchbury, F.; Richard-Loendt, A.; et al. Mutant IDH sensitizes gliomas to endoplasmic reticulum stress and triggers apoptosis via miR-183-mediated inhibition of semaphorin 3E. Cancer Res. 2019, 79, 4994–5007. [Google Scholar] [CrossRef] [Green Version]
- Li, Y.L.; Hu, X.; Li, Q.Y.; Wang, F.; Zhang, B.; Ding, K.; Tan, B.Q.; Lin, N.M.; Zhang, C. Shikonin sensitizes wild-type EGFR NSCLC cells to erlotinib and gefitinib therapy. Mol. Med. Rep. 2018, 18, 3882–3890. [Google Scholar] [CrossRef] [Green Version]
- Liao, C.H.; Tzeng, Y.T.; Lai, G.M.; Chang, C.L.; Hu, M.H.; Tsai, W.L.; Liu, Y.R.; Hsia, S.; Chuang, S.E.; Chiou, T.J.; et al. Omega-3 fatty acid-enriched fish oil and selenium combination modulates endoplasmic reticulum stress response elements and reverses acquired gefitinib resistance in HCC827 lung adenocarcinoma cells. Mar. Drugs 2020, 18, 399. [Google Scholar] [CrossRef]
- Chang, C.Y.; Shen, C.C.; Su, H.L.; Chen, C.J. Gefitinib induces apoptosis in human glioma cells by targeting Bad phosphorylation. J. Neurooncol. 2011, 105, 507–522. [Google Scholar] [CrossRef]
- Chang, C.Y.; Kuan, Y.H.; Ou, Y.C.; Li, J.R.; Wu, C.C.; Pan, P.H.; Chen, W.Y.; Huang, H.Y.; Chen, C.J. Autophagy contributes to gefitinib-induced glioma cell growth inhibition. Exp. Cell Res. 2014, 327, 102–112. [Google Scholar] [CrossRef]
- Chang, C.Y.; Li, J.R.; Wu, C.C.; Ou, Y.C.; Chen, W.Y.; Kuan, Y.H.; Wang, W.Y.; Chen, C.J. Valproic acid sensitizes human glioma cells to gefitinib-induced autophagy. IUBMB Life 2015, 67, 869–879. [Google Scholar] [CrossRef] [Green Version]
- Zhang, H.; Nakajima, S.; Kato, H.; Gu, L.; Yoshitomi, T.; Nagai, K.; Shinmori, H.; Kokubo, S.; Kitamura, M. Selective, potent blockade of the IRE1 and ATF6 pathways by 4-phenylbutyric acid analogues. Br. J. Pharmacol. 2013, 170, 822–834. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lee, H.Y.; Kim, H.K.; Hoang, T.H.; Yang, S.; Kim, H.R.; Chae, H.J. The correlation of IRE1α oxidation with Nox4 activation in aging-associated vascular dysfunction. Redox Biol. 2020, 7, 101727. [Google Scholar] [CrossRef] [PubMed]
- Zhao, Q.; Zhong, J.; Bi, Y.; Liu, Y.; Liu, Y.; Guo, J.; Pan, L.; Tan, Y.; Yu, X. Gambogenic acid induces Noxa-mediated apoptosis in colorectal cancer through ROS-dependent activation of IRE1alpha/JNK. Phytomedicine 2020, 78, 153306. [Google Scholar] [CrossRef] [PubMed]
- Rodvold, J.J.; Xian, S.; Nussbacher, J.; Tsui, B.; Cameron Waller, T.; Searles, S.C.; Lew, A.; Jiang, P.; Babic, I.; Nomura, N.; et al. IRE1alpha and IGF signaling predict resistance to an endoplasmic reticulum stress-inducing drug in glioblastoma cells. Sci. Rep. 2020, 10, 8348. [Google Scholar] [CrossRef] [PubMed]
- Auf, G.; Jabouille, A.; Delugin, M.; Guérit, S.; Pineau, R.; North, S.; Platonova, N.; Maitre, M.; Favereaux, A.; Vajkoczy, P.; et al. High epiregulin expression in human U87 glioma cells relies on IRE1α and promotes autocrine growth through EGF receptor. BMC Cancer 2013, 13, 597. [Google Scholar] [CrossRef] [PubMed]
- Dadey, D.Y.; Kapoor, V.; Khudanyan, A.; Urano, F.; Kim, A.H.; Thotala, D.; Hallahan, D.E. The ATF6 pathway of the ER stress response contributes to enhanced viability in glioblastoma. Oncotarget 2016, 7, 2080–2092. [Google Scholar] [CrossRef] [Green Version]
- Hou, X.; Liu, Y.; Liu, H.; Chen, X.; Liu, M.; Che, H.; Guo, F.; Wang, C.; Zhang, D.; Wu, J.; et al. PERK silence inhibits glioma cell growth under low glucose stress by blockage of p-AKT and subsequent HK2’s mitochondria translocation. Sci. Rep. 2015, 5, 9065. [Google Scholar] [CrossRef] [Green Version]
- Le Reste, P.J.; Pineau, R.; Voutetakis, K.; Samal, J.; Jégou, G.; Lhomond, S.; Gorman, A.M.; Samali, A.; Patterson, J.B.; Zeng, Q.; et al. Local intracerebral inhibition of IRE1 by MKC8866 sensitizes glioblastoma to irradiation/chemotherapy in vivo. Cancer Lett. 2020, 494, 73–83. [Google Scholar] [CrossRef]
- Paiva, C.; Godbersen, J.C.; Soderquist, R.S.; Rowland, T.; Kilmarx, S.; Spurgeon, S.E.; Brown, J.R.; Srinivasa, S.P.; Danilov, A.V. Cyclin-dependent kinase inhibitor P1446A induces apoptosis in a JNK/p38 MAPK-dependent manner in chronic lymphocytic leukemia B-cells. PLoS ONE 2015, 10, e0143685. [Google Scholar] [CrossRef] [PubMed]
- Quan, J.H.; Gao, F.F.; Lee, M.; Yuk, J.M.; Cha, G.H.; Chu, J.Q.; Wang, H.; Lee, Y.H. Involvement of endoplasmic reticulum stress response and IRE1-mediated ASK1/JNK/Mcl-1 pathways in silver nanoparticle-induced apoptosis of human retinal pigment epithelial cells. Toxicology 2020, 442, 152540. [Google Scholar] [CrossRef]
- Foster, K.A.; Jane, E.P.; Premkumar, D.R.; Morales, A.; Pollack, I.F. NVP-BKM120 potentiates apoptosis in tumor necrosis factor-related apoptosis-inducing ligand-resistant glioma cell lines via upregulation of Noxa and death receptor 5. Int. J. Oncol. 2015, 47, 506–516. [Google Scholar] [CrossRef] [Green Version]
- Liu, X.; Zhao, P.; Wang, X.; Wang, L.; Zhu, Y.; Gao, W. Triptolide induces glioma cell autophagy and apoptosis via upregulating the ROS/JNK and downregulating the Akt/mTOR signaling pathways. Front. Oncol. 2019, 9, 387. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Orcutt, K.P.; Parsons, A.D.; Sibenaller, Z.A.; Scarbrough, P.M.; Zhu, Y.; Sobhakumari, A.; Wilke, W.W.; Kalen, A.L.; Goswami, P.; Miller, F.J., Jr.; et al. Erlotinib-mediated inhibition of EGFR signaling induces metabolic oxidative stress through NOX4. Cancer Res. 2011, 71, 3932–3940. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Gupta, P.; Jagavelu, K.; Mishra, D.P. Inhibition of NADPH oxidase-4 potentiates 2-deoxy-d-glucose-induced suppression of glycolysis, migration, and invasion in glioblastoma cells: Role of the Akt/HIF1α/HK-2 signaling axis. Antioxid. Redox Signal. 2015, 23, 665–681. [Google Scholar] [CrossRef] [PubMed]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 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
Chang, C.-Y.; Pan, P.-H.; Wu, C.-C.; Liao, S.-L.; Chen, W.-Y.; Kuan, Y.-H.; Wang, W.-Y.; Chen, C.-J. Endoplasmic Reticulum Stress Contributes to Gefitinib-Induced Apoptosis in Glioma. Int. J. Mol. Sci. 2021, 22, 3934. https://doi.org/10.3390/ijms22083934
Chang C-Y, Pan P-H, Wu C-C, Liao S-L, Chen W-Y, Kuan Y-H, Wang W-Y, Chen C-J. Endoplasmic Reticulum Stress Contributes to Gefitinib-Induced Apoptosis in Glioma. International Journal of Molecular Sciences. 2021; 22(8):3934. https://doi.org/10.3390/ijms22083934
Chicago/Turabian StyleChang, Cheng-Yi, Ping-Ho Pan, Chih-Cheng Wu, Su-Lan Liao, Wen-Ying Chen, Yu-Hsiang Kuan, Wen-Yi Wang, and Chun-Jung Chen. 2021. "Endoplasmic Reticulum Stress Contributes to Gefitinib-Induced Apoptosis in Glioma" International Journal of Molecular Sciences 22, no. 8: 3934. https://doi.org/10.3390/ijms22083934