为了探究整合因子复合物亚基10(integrator complex subunits 10,INTS10)对人肝细胞癌(hepatocellular carcinoma,HCC)细胞周期、凋亡、生长和迁移能力的影响及其潜在的分子作用机制,利用慢病毒感染法获得稳定过表达或敲低INTS10的HCC细胞系,采用qRT-PCR和 Western blotting检测INTS10 mRNA和蛋白表达水平,接着采用CCK-8法、克隆形成和BrdU实验检测细胞生长情况,采用Tr-answell 小室实验检测细胞迁移能力,采用流式分析术检测细胞的周期和凋亡.结果显示:过表达INTS10可显著抑制HCC细胞的凋亡、生长和迁移能力,促进G1期细胞数量的增加,而敲低INTS10则呈现相反的表型.通过通路富集分析发现,周期相关通路被显著富集,过表达INTS10后,CDC25A和CDK4的mRNA和蛋白质水平显著减少,而CDKN1A的水平显著增加,敲低INTS10则呈现相反趋势.综上,本研究初步揭示了 INTS10在HCC细胞中可能通过影响G1/S期相关蛋白质的表达而发挥抑癌基因的功能,为下一步更为深入的功能和机制研究提供了基础.
Effect of INTS10 on cell cycle,apoptosis,growth and migration capacity of HCC
The effects of integrator complex subunits 10(INTS10)on cell cycle,apoptosis,growth and migration of hepatocellular carcinoma(HCC)cells and its potential molecular mechanism were explored.The HCC cell lines with stable overexpression or knockdown of INTS10 were obtained by lentivirus infection.The mRNA and protein expression levels of INTS10 were detected by qRT-PCR and Western blotting.The cell growth was detected by CCK-8,clonal formation and BrdU assay.The cell migration ability was detected by Transwell assay.Flow cytometry was used to detect the number of cell cycles and apoptosis.The results showed that overexpression of INTS10 could significantly inhibit the apoptosis,growth and migration of HCC,and promote the increase of the number of G1 phase cells,while knockdown of INTS10 showed the opposite effects.Through pathway enrichment analysis,it was found that the cycle-dependent pathway was significantly enriched.Overexpression of INTS10 could significantly reduce the mRNA and protein levels of CDC25A and CDK4,while the level of CDKN1A was significantly increased,and INTS10 knockdown showed the opposite trend.In conclusion,it is preliminarily revealed that INTS10 may play a role as a tumor suppressor gene in HCC by affecting the expression of G1/S phase related proteins,which provides a foundation for further functional and mechanism studies.
万方数据
维普
