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,杨凌博,魏澎涛,李小辉,孙建涛,杨锦建
中国临床解剖学杂志 ›› 2018, Vol. 36 ›› Issue (6) : 615-620.
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
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。
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.
膀胱癌 / miR-210-3p / autophagy-related gene 7 / 自噬
/ Bladder cancer / miR-210-3p / Autophagy-related gene 7 / Autophagy
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国家自然科学基金（81570685）
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