Rab1A siRNA对宫颈癌HeLa细胞增殖和凋亡的影响及其分子机制

Effects of Rab1A siRNA on proliferation and apoptosis of cervical cancer He La cell line and its molecular mechanism

目的观察Rab1A对宫颈癌He La细胞增殖和凋亡的影响并探讨其分子机制。方法应用siRNA干扰Rab1A基因表达,将He La细胞分为空白对照组、阴性对照组和Rab1A siRNA组,空白对照组不做处理,阴性对照组转染阴性对照siRNA。Rab1A siRNA组转染Rab1A特异性siRNA。MTT比色法分析Rab1A对宫颈癌He La细胞增殖的影响;采用流式细胞仪分析对细胞周期和细胞凋亡的影响;Western blot观察Rab1A siRNA对Cyclin D1、GRP78、Bcl-2和Bax表达的影响。结果与阴性对照组相比,Rab1A siRNA组He La细胞的Rab1A mRNA和蛋白的表达均显著下调;Rab1A siRNA抑制宫颈癌He La细胞增殖;与阴性对照组相比,Rab1A siRNA组He La细胞的S期细胞数量显著减少(P<0.05),G1/G0期细胞数量显著增加(P<0.05),Rab1A siRNA组He La细胞的早期凋亡和晚期凋亡显著增加(P<0.01);Rab1A siRNA组与阴性对照组相比,Cyclin D1和Bcl-2在蛋白水平的表达显著下调(P<0.01),GRP78和Bax在蛋白水平的表达显著上调(P<0.01)。结论 Rab1A通过调控Cyclin D1的表达促进宫颈癌He La细胞增殖,通过调控GRP78、Bcl-2和Bax表达抑制细胞凋亡。

Objective To investigate the effects of Rab1 A on the proliferation and apoptosis of cervical cancer He La cell line and its molecular mechanism. Methods The siRNA interference was used to knockdown Rab1 A. He La cells were divided into three groups： blank group,negative control group and Rab1 A siRNA group. Control siRNA and Rab1A-targeted siRNA were transfected into He La cells in negative control group and Rab1 A siRNA group,respectively. MTT assay was used to analyze the effects of Rab1 A on proliferation of cervical cancer He La cells. The effects of Rab1 A on the cell cycle and apoptosis of cervical cancer He La cells were observed by flow cytometry. The effects of Rab1 A siRNA on the expression of Cyclin D1,GRP78,Bcl-2 and Bax were analyzed by Western blot. Results Rab1 A mRNA and protein expression significantly decreased in Rab1 A siRNA group compared to negative control group（ P〈0. 01）. Rab1 A siRNA inhibited the proliferation of cervical cancer He La cells. The number of S phase cells significantly decreased in Rab1 A siRNA group compared to negative control group（ P〈0. 05）,meanwhile the number of G1/ G0 phase cells significantly increased（ P〈0. 05）. The number of early apoptotic cells and late apoptotic cells significantly increased in Rab1 A siRNA group compared to negative control group（ P〈0. 01）. Cyclin D1 and Bcl-2 protein expression significantly decreased in Rab1 A siRNA group compared to negative control group（ P〈0. 01）,while GRP78 and Bax protein expression remarkably increased（ P〈0. 01）. Conclusion Rab1 A may promote the proliferation of cervical cancer He La cells through regulating Cyclin D1 expression,and inhibit the apoptosis by GRP78,Bcl-2 and Bax expression.