The Role of endoplasmic reticulum metallo protease 1 on Autophagy Pathway in HCT-116 Colorectal Cancer Cell Line

Document Type : Research/Original Article

Authors

1 Colorectal Research Center, Shiraz University of Medical Sciences, Shiraz, Iran Autophagy Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

2 Endocrinology and Metabolism Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

3 Department of Biochemistry, Faculty of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran Department of Medical Laboratory Sciences, Faculty of Medical Sciences, Islamic Azad University, Arak Branch, Arak, Iran

4 Colorectal Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

5 Autophagy Research Center, Shiraz University of Medical Sciences, Shiraz, Iran Department of Biochemistry, Faculty of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

Abstract

Background: Autophagy and unfolded protein response (UPR) are mechanisms with dual roles in both maintaining the cellular homeostasis and progression of various diseases such as cancer. Therefore, identification of different molecules and proteins involved in the regulation of these pathways may contribute to find new therapeutic targets. A member of the M28 family of the metallopeptidases, Endoplasmic Reticulum Metallo Protease 1 (ERMP1), is overexpressed in cancers such as colorectal cancer. The role of this protein in the UPR activation was previously reported in breast cancer. We aimed to evaluate the role of ERMP1 in the activation of autophagy and apoptosis in colorectal cancer. Methods: ERMP1 Gene silencing was performed using specific small hairpin RNA (shRNA) in HCT-116 colorectal cancer cell line. Then, autophagy associated protein markers including Beclin 1, p62 and LC3II were evaluated using western blot. The effect of ERMP1 knockdown on cellular apoptosis was also assessed by propidium iodide staining flow cytometry analysis. Statistical analysis was performed using SPSS software version 20. Results: All three autophagy markers were increased significantly in the ERMP1-silenced HCT116 cell lines compared with negative control cells (P < 0.05). It seems that ERMP1 silencing inhibits autophagy at the flux stage. However, ERMP1 knockdown had no significant effect on HCT-116 apoptotic cell death (P > 0.05). Conclusion: The oncogenic protein, ERMP1, activates autophagy in colorectal cancer cell line. Targeting of ERMP1 may be considered as a proper approach in colorectal cancer therapy. Further investigations are required to confirm these results.

Keywords

References

1.            Koveitypour Z, Panahi F, Vakilian M, Peymani M, Forootan FS, Esfahani MHN, et al. Signaling pathways involved in colorectal cancer progression. Cell & bioscience. 2019;9(1):1-14.
2.            Amirghofran Z, Jalali SA, Ghaderi A, Hosseini SV. Genetic polymorphism in the transforming growth factor β1 gene (-509 C/T and-800 G/A) and colorectal cancer. Cancer genetics and cytogenetics. 2009;190(1):21-5.
3.            Mathew R, Karantza-Wadsworth V, White E. Role of autophagy in cancer. Nature Reviews Cancer. 2007;7(12):961-7.
4.            Yu L, Chen Y, Tooze SA. Autophagy pathway: cellular and molecular mechanisms. Autophagy. 2018;14(2):207-15.
5.            Zhang Y, Zhang Y, Jin X-f, Zhou X-h, Dong X-h, Yu W-t, et al. The role of astragaloside IV against cerebral ischemia/reperfusion injury: suppression of apoptosis via promotion of P62-LC3-autophagy. Molecules. 2019;24(9):1838.
6.            Lippai M, Lőw P. The role of the selective adaptor p62 and ubiquitin-like proteins in autophagy. BioMed research international. 2014;2014.
7.            Liu WJ, Ye L, Huang WF, Guo LJ, Xu ZG, Wu HL, et al. p62 links the autophagy pathway and the ubiqutin–proteasome system upon ubiquitinated protein degradation. Cellular & molecular biology letters. 2016;21(1):1-14.
8.            Song S, Tan J, Miao Y, Zhang Q. Crosstalk of ER stress‐mediated autophagy and ER‐phagy: Involvement of UPR and the core autophagy machinery. Journal of cellular physiology. 2018;233(5):3867-74.
9.            Suh DH, Kim M-K, Kim HS, Chung HH, Song YS. Unfolded protein response to autophagy as a promising druggable target for anticancer therapy. Annals of the New York Academy of Sciences. 2012;1271(1):20.
10.          Xie W, Zhou J. Aberrant regulation of autophagy in mammalian diseases. Biology letters. 2018;14(1):20170540.
11.          Singh SS, Vats S, Chia AY-Q, Tan TZ, Deng S, Ong MS, et al. Dual role of autophagy in hallmarks of cancer. Oncogene. 2018;37(9):1142-58.
12.          Choi KS. Autophagy and cancer. Experimental & molecular medicine. 2012;44(2):109-20.
13.          Mele L, Del Vecchio V, Liccardo D, Prisco C, Schwerdtfeger M, Robinson N, et al. The role of autophagy in resistance to targeted therapies. Cancer Treatment Reviews. 2020;88.
14.          Chen N, Karantza-Wadsworth V. Role and regulation of autophagy in cancer. Biochimica et Biophysica Acta (BBA)-Molecular Cell Research. 2009;1793(9):1516-23.
15.          Amaravadi RK, Lippincott-Schwartz J, Yin X-M, Weiss WA, Takebe N, Timmer W, et al. Principles and current strategies for targeting autophagy for cancer treatment. Clinical cancer research. 2011;17(4):654-66.
16.          Yang H-Z, Ma Y, Zhou Y, Xu L-M, Chen X-J, Ding W-B, et al. Autophagy contributes to the enrichment and survival of colorectal cancer stem cells under oxaliplatin treatment. Cancer letters. 2015;361(1):128-36.
17.          Li J, Hou N, Faried A, Tsutsumi S, Kuwano H. Inhibition of autophagy augments 5-fluorouracil chemotherapy in human colon cancer in vitro and in vivo model. European journal of cancer. 2010;46(10):1900-9.
18.          Garcia-Rudaz C, Luna F, Tapia V, Kerr B, Colgin L, Galimi F, et al. Fxna, a novel gene differentially expressed in the rat ovary at the time of folliculogenesis, is required for normal ovarian histogenesis. Development. 2007;134(5):945-57.
19.          Wu J, Liu S, Liu G, Dombkowski A, Abrams J, Martin-Trevino R, et al. Identification and functional analysis of 9p24 amplified genes in human breast cancer. Oncogene. 2012;31(3):333-41.
20.          Grandi A, Santi A, Campagnoli S, Parri M, De Camilli E, Song C, et al. ERMP1, a novel potential oncogene involved in UPR and oxidative stress defense, is highly expressed in human cancer. Oncotarget. 2016;7(39):63596.
21.          Kuroda H, Kutner RH, Bazan NG, Reiser J. Simplified lentivirus vector production in protein-free media using polyethylenimine-mediated transfection. Journal of virological methods. 2009;157(2):113-21.
22.          Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991;139(2):271-9.
23.          Chen X-l, Liu P, Zhu W-l, Lou L-g. DCZ5248, a novel dual inhibitor of Hsp90 and autophagy, exerts antitumor activity against colon cancer. Acta Pharmacologica Sinica. 2021;42(1):132-41.
24.          Qu J, Zhang L, Li L, Su Y. MiR-148b functions as a tumor suppressor by targeting endoplasmic reticulum metallo protease 1 in human endometrial cancer cells. Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics. 2018;27(1):81-8.
25.          Deegan S, Koryga I, Glynn SA, Gupta S, Gorman AM, Samali A. A close connection between the PERK and IRE arms of the UPR and the transcriptional regulation of autophagy. Biochemical and biophysical research communications. 2015;456(1):305-11.
26.          Antonioli M, Di Rienzo M, Piacentini M, Fimia GM. Emerging mechanisms in initiating and terminating autophagy. Trends in biochemical sciences. 2017;42(1):28-41.
27.          Pugsley HR. Assessing autophagic flux by measuring LC3, p62, and LAMP1 co-localization using multispectral imaging flow cytometry. Journal of visualized experiments: JoVE. 2017(125).
Iranian Journal of Colorectal Research
Volume 9, Issue 2
June 2021
Pages 63-68

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