Inhibition of Autophagy on Melatonin‑Induced Apoptosis in MCF-7 and MDA-MB-231 Cell Lines

Yıl 2024, Cilt: 6 Sayı: 1, 107 - 114, 31.01.2024

Gözde Özge Önder , Adnan Bayram , Oğuz Galip Yıldız , Arzu Yay

https://doi.org/10.37990/medr.1400485

Öz

Aim: We looked at the connection between autophagy and apoptosis after our prior research indicated that melatonin could cause MCF-7 and MDA-MB-231 cells in the present study.
Material and Method: In order to investigate the autophagy inhibition’s effect on the melatonin-induced BC cells’ apoptosis, melatonin and/or 3-methyladenine (3-MA, autophagy inhibitor) have been utilized. Melatonin was applied to the cells following a 5-mM 3-MA pre-cultivation. Then, apoptosis was detected by the TUNEL method. The technique for double immunofluorescence labeling was used to identify the molecular alterations in Bax/Bcl-2 expression. To evaluate the cell viability, the MTT test was used.
Results: When an autophagy inhibitor, 3-MA, and melatonin treatment were co-administered in MCF-7 cells, apoptosis was decreased, compared to melatonin treatment alone, but it was not significant. In addition, 3-MA application downregulated Bax expression compared with melatonin alone treatment. Combined therapy markedly elevated apoptosis and significantly up-regulated Bax protein in MDA-MB-231 cells.
Conclusion: Taken together, in MCF-7 cells, autophagy’s inhibition contributes to the downregulation of apoptosis, whereas increased apoptosis is seen in MDA-MB-231 cells. Inhibiting autophagy in these cells treated with melatonin could serve as a self-defense mechanism, and This might be a good strategy for breast cancer adjuvant treatment.

Anahtar Kelimeler

Autophagy , Breast cancer , Melatonin , 3-MA

Destekleyen Kurum

Health Sciences Research Support Group (SBAG) of TUBITAK

Proje Numarası

220S879

Kaynakça

• Gluz O, Liedtke C, Gottschalk N, et al. Triple-negative breast cancer-current status and future directions. Ann Oncol. 2009;20:1913-27.
• Murphy CG, Dickler MN. Endocrine resistance in hormone responsive breast cancer: mechanisms and therapeutic strategies. Endocr Relat Cancer. 2016;23:R337-52.
• Wang C, Kar S, Lai X, et al. Triple negative breast cancer in Asia: an insider’s view. Cancer Treat Rev. 2018;62:29-8.
• La porta E, Welsh J. Modelling vitamin D in triple negative/basal likebreast cancer-Review. J Steroid Biochem Mol Bio. 2014;144:65-73.
• Comsa S, Cimpean AM, Raica M. The story of MCF-7 breast cancer cell line: 40 years of experience in research. Anticancer Res. 2015;35:3147-54.
• Theodossiou TA, Ali M, Grigalavicius M, et al. Simultaneous defeat of MCF7 and MDA-MB-231 resistances by a hypericin PDT–tamoxifen hybrid therapy. NPJ Breast Cancer. 2019;5:13.
• Yin L, Duan JJ, Bian XW, Yu SC. Triple-negative breast cancer molecular subtyping and treatment progress. Breast Cancer Res. 2020;22:61.
• da Silva JL, Cardoso Nunes NC, Izetti P, et al. Triple negative breast cancer: a thorough review of biomarkers. Crit Rev Oncol Hematol. 2020;145:102855.
• Reis-Filho JS, Pusztai L. Gene expression profiling in breast cancer: Classification, prognostication, and prediction. Lancet. 2011;378:1812-23.
• González-Martínez S, Pérez-Mies B, Carretero-Barrio I, et al. Molecular features of metaplastic breast carcinoma: an infrequent subtype of triple negative breast carcinoma. Cancers. 2020;12:1832.
• Acuña-Castroviejo D, Escames G, Venegas C, et al. Extrapineal melatonin: sources, regulation, and potential functions. Cell Mol Life Sci. 2014;71:2997-3025.
• Luchetti F, Canonico B, Betti M, et al. Melatonin signaling and cell protection function. FASEB J. 2010;24:3603-24.
• Sainz RM, Mayo JC, Tan DX, et al. Melatonin reduces prostate cancer cell growth leading to neuroendocrine differentiation via a receptor and PKA independent mechanism. Prostate. 2005;63:29-43.
• Sainz RM, Mayo JC, Tan DX, et al. Antioxidant activity of melatonin in Chinese hamster ovarian cells: changes in cellular proliferation and differentiation. Biochem Biophys Res Commun. 2003;302:625-34.
• Bizzarri M, Proietti S, Cucina A, Reiter RJ. Molecular mechanisms of the pro-apoptotic actions of melatonin in cancer: a review. Expert Opin Therap Targets. 2013;17:1483-96.
• Juhasz G, Sass M. Hid can induce, but is not required for autophagy in polyploid larval Drosophila tissues. Eur Cell Biol. 2005;84:491-502.
• Wu Z, Wu LJ, Li LH, et al. p53- mediated cell cycle arrest and apoptosis induced by shikonin via a caspase-9-dependent mechanism in human malignant melanoma A375-S2 cells. J Pharmacol Sci. 2004;94:166-76.
• Mizushima N, Levine B, Cuervo AM, Klionsky DJ. Autophagy fights disease through cellular self-digestion. Nature. 2008;451:1069-75.
• Han X, Mo J, Yang Y, et al. Crucial roles of LncRNAs-mediated autophagy in breast cancer. Int J Med Sci. 2022;19:1082-92.
• Petiot A, Ogier-Denis E, Blommaart EFC, et al. Distinct classes of phosphatidylinositol 30 kinases are involved in signaling pathways that control macroautophagy in HT–29 cells. J Biol Chem. 2000;275:992-8.
• Pan X, Zhang X, Sun H, et al. Autophagy inhibition promotes 5-fluorouraci-induced apoptosis by stimulating ROS formation in human non-small cell lung cancer A549 cells. PLoS One. 2013;8:e56679.
• Kumar D, Shankar S, Srivastava RK. Rottlerin induces autophagy and apoptosis in prostate cancer stem cells via PI3K/Akt/ mTOR signaling pathway. Cancer Lett. 2013;343:179-89.
• Li J, Hou N, Faried A, et al. Inhibition of autophagy by 3-MA enhances the effect of 5-FU-induced apoptosis in colon cancer cells. Ann Surg Oncol. 2009;16:761-71.
• Cui Q, Tashiro S, Onodera S, Ikejima T. Augmentation of oridonin-induced apoptosis observed with reduced autophagy. J Pharmacol Sci. 2006;101:230-9.
• Cui Q, Tashiro S, Onodera S, et al. Autophagy preceded apoptosis in oridonin-treated human breast cancer MCF-7 cells. Biol Pharm Bull. 2007;30:859-64.
• Onder GO, Sezer G, Ozdamar S, Yay A. Melatonin has an inhibitory effect on MCF-7 and MDA-MB-231 human breast cancer cell lines by inducing autophagy and apoptosis. Fund Clin Pharmacol. 2022;36:1038-56.
• Kroemer G, Jaattela M. Lysosomes and autophagy in cell death control. Nat Rev Cancer. 2005;5:886-97.
• Kondo Y, Kondo S. Autophagy and cancer therapy. Autophagy. 2006;2:85-90.
• Kroemer G, Levine B. Autophagic cell death: the story of a misnomer. Nat Rev Mol Cell Biol. 2008;9:1004-10.
• Eisenberg-Lerner A, Bialik S, Simon HU, Kimchi A. Life and death partners: apoptosis, autophagy and the cross-talk between them. Cell Death Differ. 2009;16:966-75.
• Tran AT, Ramalinga M, Kedir H, et al. Autophagy inhibitor 3-methyladenine potentiates apoptosis induced by dietary tocotrienols in breast cancer cells. Eur J Nutr. 2015;54:265-72.
• Chen S, Gao Y, Zhu P, et al. Anti-cancer drug anlotinib promotes autophagy and apoptosis in breast cancer. Front Biosci (Landmark Ed). 2022;27:125-38.
• Han SH, Lee JH, Woo JS, et al. Myricetin induces apoptosis through the MAPK pathway and regulates JNK‑mediated autophagy in SK‑BR‑3 cells. Int J Mol Med. 2022;49:54-65.