miR-210-3p 通过靶向ATG7 抑制人膀胱癌细胞J82的自噬和诱导凋亡

doi:10.13418/j.issn.1001-165x.2018.06.004

中国临床解剖学杂志 ›› 2018, Vol. 36 ›› Issue (6): 615-620.doi: 10.13418/j.issn.1001-165x.2018.06.004

miR-210-3p 通过靶向ATG7 抑制人膀胱癌细胞J82的自噬和诱导凋亡

韩兴涛¹，　杨凌博¹，　魏澎涛¹，　李小辉¹，　孙建涛¹，　杨锦建²

1.洛阳市中心医院泌尿外科，河南洛阳 471000；　2.郑州大学一附院泌尿外科，郑州 450001

收稿日期:2018-05-11 出版日期:2018-11-25 发布日期:2018-12-29
作者简介:韩兴涛（1979-），硕士，副主任医师，主要从事泌尿外科方面的治疗与研究，Tel：13103796780，E-mail：ydezgp1@sina.com
基金资助:
国家自然科学基金（81570685）

miR-210-3p suppresses autophagy and promotes apoptosis by targeting autophagy-related gene 7 in bladder cancer J82 cells

HAN Xing-tao¹, YANG Ling-bo¹, WEI Peng-tao¹, LI Xiao-hui¹, SUN Jian-tao¹, YANG Jin-jian²

1. Department of Urology, Luoyang Central Hospital, Luoyang 471000, Henan Province, China; 2. Department of Urology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450001, China

Received:2018-05-11 Online:2018-11-25 Published:2018-12-29

摘要/Abstract

摘要：

目的　探索miR-210-3p抑制人膀胱癌细胞J82机制。方法　应用定量实时PCR（qRT-PCR）检测miR-210-3p及其靶基因自噬相关基因7 （autophagy-related gene 7，ATG7）在J82细胞中的表达。为验证miR-210-3p和ATG7之间的生物学关系，进行荧光素酶报告检测。通过体外实验研究miR-210-3p和ATG7在J82细胞中的生物学功能，包括CCK-8法检测细胞增殖情况，hoechst染色检测细胞凋亡，western blot检测细胞自噬。结果　miR-210-3p表达明显下调ATG7表达水平，生物信息学预测和荧光素酶报告实验证明miR-210-3p抑制ATG7是通过直接结合到ATG7 3’ -未翻译区域(3’- UTR)。在J82细胞中，miR-210-3p上调可抑制ATG7表达、细胞自噬和细胞增殖，同时诱导细胞凋亡。结论　miR-210-3p通过负调控ATG7抑制细胞增殖和自噬，并促进凋亡，从而促进膀胱癌细胞J82细胞死亡。因此，在膀胱癌中抑制自噬对于开发针对膀胱癌的自噬靶向治疗至关重要。本研究补充了miRNA调控膀胱癌的相关机制。

关键词: 膀胱癌, 　miR-210-3p, 　autophagy-related gene 7, 　自噬

Abstract:

Objective　This paper mainly explored the regulatory mechanism of miR-210-3p in bladder cancer progression in J82 cell.　Methods　Quantitative real-time PCR (qRT-PCR) was applied to detect the expression of miR-210-3p and its target gene autophagy related gene 7 (ATG7) in J82 cells. The luciferase reporter assay was conducted to verify the biological relationship between miR-210-3p and ATG7. The biological functions of miR-210-3p and ATG7 in J82 cells were studied by in vitro experiments (CCK-8, hoechst staining, and Western blot). Results miR-210-3p expression significantly reduced the ATG7 expression level. Bioinformatics prediction and luciferase reporter assay demonstrated that miR-23-3p inhibited ATG7 by directly binding to ATG7 3’UTR. In J82 cells, miR-210-3p overexpression inhibited ATG7 expression, cell autophagy and cell proliferation, and induced cell apoptosis.　Conclusions　mir-210-3p can inhibit J82 cell proliferation, autophagy and promote apoptosis through negative regulation of ATG7, thereby promoting the death of cell J82 cells. Therefore, inhibition of autophagy in bladder cancer is essential for the development of autophagy targeting therapy for bladder cancer. This study further supports the mechanism of miR-210-3p in regulation of bladder cancer.

Key words: , Bladder cancer, 　miR-210-3p, 　Autophagy-related gene 7, 　Autophagy

韩兴涛,杨凌博,魏澎涛,李小辉,孙建涛,杨锦建. miR-210-3p 通过靶向ATG7 抑制人膀胱癌细胞J82的自噬和诱导凋亡[J]. 中国临床解剖学杂志, 2018, 36(6): 615-620.

HAN Xing-tao, YANG Ling-bo, WEI Peng-tao, LI Xiao-hui, SUN Jian-tao, YANG Jin-jian. miR-210-3p suppresses autophagy and promotes apoptosis by targeting autophagy-related gene 7 in bladder cancer J82 cells[J]. Chinese Journal Of Clinical Anatomy, 2018, 36(6): 615-620.

参考文献

[1] Afshari M, Janbabaei G, Bahrami MA, et al. Opium and bladder cancer: A systematic review and meta-analysis of the odds ratios for opium use and the risk of bladder cancer[J]. Plos One, 2017, 12(6):e0178527.
[2] Gee HE, Camps C, Buffa FM, et al. Hsa-mir-210 is a marker of tumor hypoxia and a prognostic factor in head and neck cancer[J]. Cancer, 2010, 116(9): 2148-2158.
[3] Alan Ka-Lun Kai, Lo Kong Chan, Regina Cheuk-Lam Lo, et al. Down-regulation of timp2 by hif-1alpha/mir-210/hif-3alpha regulatory feedback circuit enhances cancer metastasis in hepatocellular carcinoma[J]. Hepatology (Baltimore, Md), 2016, 64(2): 473-487.
[4] Chang Y, Yan W, He X, et al. miR-375 inhibits autophagy and reduces viability of hepatocellular carcinoma cells under hypoxic conditions[J]. Gastroenterology, 2012, 143(12): 177-187.
[5] Ho AS, Huang X, Cao HB, et al. Circulating mir-210 as a novel hypoxia marker in pancreatic cancer[J]. Translational Oncology, 2010, 3(2): 109-113.
[6] Xiaoming Yang, Lei Shi, Chengzhi Yi, et al. Mir-210-3p inhibits the tumor growth and metastasis of bladder cancer via targeting fibroblast growth factor receptor-like 1[J]. Am J Cancer Res, 2017, 7(8): 1738-1753.
[7] Yuchen Liu, Yonghua Han, Hu Zhang, et al. Synthetic mirna-mowers targeting mir-183-96-182 cluster or mir-210 inhibit growth and migration and induce apoptosis in bladder cancer cells[J]. PloS One, 2012, 7(12): e52280.
[8] Ren D, Yang Q, Dai Y, et al. Oncogenic mir-210-3p promotes prostate cancer cell emt and bone metastasis via nf-kappab signaling pathway[J]. Mol Cancer, 2017, 16(1): 117.
[9] Lan H, Lu H, Wang X, et al. MicroRNAs as potential biomarkers in cancer: opportunities and challenges[J]. Bio Med Res Int, 2015, 2015(1): 125094.
[10]Li P, Xue WJ, Feng Y, et al. Microrna-205 functions as a tumor suppressor in colorectal cancer by targeting camp responsive element binding protein 1 (creb1)[J]. Am J Transl Res, 2015,7(10): 2053-2059.
[11]Dayue Liu, Haoming Xia, Fang Wang, et al. Microrna-210 interacts with fbxo31 to regulate cancer proliferation cell cycle and migration in human breast cancer[J]. OncoTargets Ther, 2016, 9: 5245-5255.
[12]Zhang C, Tian W, Meng L, et al. Prl-3 promotes gastric cancer migration and invasion through a nf-kappab-hif-1alpha-mir-210 axis[J]. J Mol Med (Berlin, Germany), 2016, 94(4): 401-415.
[13]Gautam Das, Bhupendra V. Shravage, Eric H. Baehrecke. Regulation and function of autophagy during cell survival and cell death[J]. Cold Spring Harb Perspect Biol, 2012, 4(6): 997-1001.
[14]L Galluzzi, JM Bravo-San Pedro, I Vitale, et al. Essential versus accessory aspects of cell death: recommendations of the nccd 2015[J]. Cell Death Differ, 2015, 22(1): 58-73.
[15]Juliet Goldsmith, Beth Levine, Jayanta Debnath. Autophagy and cancer metabolism[J]. Methods Enzymol, 2014, 542: 25-57.
[16] Banzhou Pan, Bing Feng, Yitian Chen, et al. Mir-200b regulates autophagy associated with chemoresistance in human lung adenocarcinoma[J]. Oncotarget, 2015, 6(32): 32805-32820.
[17]Wang IK, Sun KT, Tsai TH, et al. Mir-20a-5p mediates hypoxia-induced autophagy by targeting atg16l1 in ischemic kidney injury[J]. Life Sciences, 2015, 136: 133-141.
[18]Anne M. Strohecker, Jessie Yanxiang Guo, et al. Autophagy sustains mitochondrial glutamine metabolism and growth of brafv600e-driven lung tumors[J]. Cancer Discov, 2013, 3(11): 1272-1285.
[19]Sergio Comincini, Giulia Allavena, Silvia Palumbo, et al. Microrna-17 regulates the expression of atg7 and modulates the autophagy process, improving the sensitivity to temozolomide and low-dose ionizing radiation treatments in human glioblastoma cells[J]. Cancer Biol Ther, 2013, 14(7): 574-586.
[20]Fulda S. Cross talk between cell death regulation and metabolism[J]. Methods Enzymol, 2014, 542: 81-90.
[21]Shenghong Yang, Xiaoxu Wang, Gianmarco Contino, et al. Pancreatic cancers require autophagy for tumor growth[J]. Genes Dev, 2011, 25(7): 717-729.

miR-210-3p 通过靶向ATG7 抑制人膀胱癌细胞J82的自噬和诱导凋亡

miR-210-3p suppresses autophagy and promotes apoptosis by targeting autophagy-related gene 7 in bladder cancer J82 cells

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