miR-210-3p通过下调USP10促进肝癌细胞的迁移和侵袭-现代医学

miR-210-3p通过下调USP10促进肝癌细胞的迁移和侵袭

单位：陕西省中医医院肝病科, 陕西西安 710000

分类号：R735.7

出版年·卷·期（页码）：2020·48·第九期（1190-1196）

摘要：

目的：探讨miR-210-3p对泛素特异性蛋白酶10（USP10）的调控作用及其对肝癌细胞迁移及侵袭的影响。方法：体外培养人肝永生化细胞THLE-3与肝癌细胞系MHCC97H、Hep3B、SK-HEP-1，采用qRT-PCR与Western blotting分别检测miR-210-3p、USP10的表达水平。利用脂质体转染技术分别将anti-miR-210-3p、anti-miR-NC、pcDNA-USP10、pcDNA-NC转染至肝癌Hep3B细胞，采用Transwell小室实验检测细胞迁移及侵袭能力。双荧光素酶报告实验明确USP10的3'UTR是否为miR-210-3p的直接靶点。Western blotting检测MMP2、MMP9蛋白表达。结果：miR-210-3p在肝癌细胞中的表达水平显著升高（P<0.05），而USP10的表达水平显著降低（P<0.05）；抑制miR-210-3p表达或USP10过表达可显著减少迁移细胞数及侵袭细胞数（P<0.05），抑制MMP2、MMP9表达（P<0.05）；双荧光素酶报告实验明确miR-210-3p靶定USP10的3'UTR并抑制USP10的蛋白表达；抑制USP10的表达可逆转抑制miR-210-3p的表达对Hep3B细胞迁移及侵袭的影响。结论：miR-210-3p通过下调USP10的表达从而促进肝癌细胞的迁移与侵袭。

Objective: To investigate the regulation of microRNA-210-3p (miR-210-3p) on ubiquitin-specific protease 10(USP10) and its effect on migration and invasion of hepatocellular carcinoma cells. Methods: THLE-3 cells and hepatoma cell lines MHCC97H, Hep3B and SK-HEP-1 were cultured in vitro, and the expression levels of miR-210-3p and USP10 were detected by real-time quantitative PCR (qRT-PCR) and Western blotting, respectively. Anti-miR-210-3p, anti-miR-NC, pcDNA-USP10, and pcDNA-NC were transfected into Hep3B cells by lipofection. Transwell chamber assay was used to detect cell migration and invasion. The dual luciferase reporter assay was used to demonstrate whether the 3'UTR of USP10 was a direct target of miR-210-3p. Western blotting was used to detect the expression of MMP2 and MMP9 proteins.Results: The expression level of miR-210-3p in liver cancer cells was significantly increased (P<0.05), while the expression level of USP10 was significantly decreased (P<0.05). Inhibition of miR-210-3p expression or USP10 overexpression could significantly reduce the number of migrating cells and invasion cells (P<0.05), and inhibit the expression of MMP2 and MMP9 (P<0.05). The dual luciferase report experiment confirmed that miR-210-3p could target the 3'UTR of USP10 and inhibit the protein expression of USP10. Inhibiting the expression of USP10 could reverse the effect of inhibiting the expression of miR-210-3p on Hep3B cell migration and invasion.Conclusion: miR-210-3p promotes the migration and invasion of hepatocellular carcinoma cells by down-regulating the expression of USP10.

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