南方医科大学学报 ›› 2017, Vol. 37 ›› Issue (02): 232-.

• • 上一篇    下一篇

沉默HERC4表达可抑制宫颈癌细胞的增殖、凋亡和迁移

危敏,张艳玲,陈岚,蔡翠霞,王涵多   

  • 出版日期:2017-02-20 发布日期:2017-02-20

RNA interference of HERC4 inhibits proliferation, apoptosis and migration of cervical cancer Hela cells

  • Online:2017-02-20 Published:2017-02-20

摘要: 目的探讨HERC4 对宫颈癌细胞生物学功能的影响及其分子机制。方法设计特异HERC4 siRNA 干扰序列, Lipofectamine2000法转染Hela细胞,Western blotting检测转染特异siRNA后Hela细胞HERC4蛋白的表达水平;CCK-8 法检测 转染后Hela细胞的增殖能力;AnnexinⅤ-FITC/PI双染法检测转染后Hela细胞凋亡率的变化;划痕实验检测转染后Hela细胞迁 移能力。Western blotting检测转染后Hela细胞Cyclin D1、Bcl-2表达变化。结果转染特异siRNA后Hela细胞HERC4蛋白表 达量明显下降,差异有统计学意义(P<0.01)。与对照组相比,干扰组Hela细胞生长速度明显减慢,细胞凋亡率增加,迁移能力明 显下降,差异有统计学意义(P<0.01)。转染后Cyclin D1和Bcl-2在蛋白水平的表达显著下调(P<0.01)。结论siRNA可有效降 低宫颈癌Hela细胞中HERC4的表达,并通过抑制Cyclin D1表达抑制宫颈癌Hela细胞的增殖和迁移能力,抑制Bcl-2表达增加 细胞凋亡能力。

Abstract: Objective To explore the effects of silencing HERC4 on the proliferation, apoptosis, and migration of cervical cancer cell line Hela and the possible molecular mechanisms. Methods Three HERC4-specific small interfering RNAs (siRNAs) were transfected into Hela cells, and HERC4 expression in the cells was examined with Western blotting. CCK-8 assay, annexin V-FITC/PI assay, and wound healing assay were used to assess the effect of HERC4 silencing on the proliferation, apoptosis and migration ability of Hela cells. The expression levels of cyclin D1 and Bcl-2 in the cells were detected using Western blotting. Results Transfection of siRNA-3 resulted in significantly decreased HERC4 protein expression (P<0.01). HERC4 silencing by siRNA-3 markedly suppressed the proliferation and migration of Hela cells, increased the apoptosis rate (P<0.01) and reduced the expression levels of cyclin D1 and Bcl-2 (P<0.01). Conclusion Silencing of HERC4 efficiently inhibits the proliferation, migration, and invasion of Hela cells in vitro, and the underlying mechanisms may involve the down-regulation of cyclin D1 and Bcl-2.