RNA干扰TMEM33的表达对宫颈癌细胞生长的影响

Effect of RNA interference with TMEM33 expression on cervical cancer cells

目的通过生物信息学方法探究跨膜蛋白33(transmembrane protein 33,TMEM33)在宫颈癌组织中的表达及其与预后的关系,观察TMEM33经RNA干扰后对宫颈癌细胞生长的影响。方法应用基因表达谱数据动态分析(Gene Expression Profiling Interactive Analysis,GEPIA)数据库筛选出与宫颈癌相关的TMEM33基因,设计两条特异siRNA序列进行RNA干扰实验。实验分为3组:siTMEM33#1干扰组、siTMEM33#2干扰组、阴性对照组(非特异性siRNA)。采用Western blot法检测siRNA干扰后宫颈癌SiHa/HeLa细胞中TMEM33蛋白的表达,CCK-8法检测SiHa/HeLa细胞增殖,流式细胞术检测细胞凋亡和细胞周期的变化,Western blot法检测细胞凋亡关键基因BCL-2、Bax及细胞周期蛋白CyclinD1、P21的表达变化。结果与阴性对照组相比,特异性siRNA显著下调宫颈癌SiHa/HeLa细胞中TMEM33蛋白的表达(P<0.05);宫颈癌细胞增殖明显受抑,细胞凋亡显著增加,细胞周期S时相明显减少(P<0.05);细胞凋亡关键基因BCL-2表达下降而Bax表达上升,细胞周期蛋白CyclinD1水平降低而P21水平升高(均为P<0.05)。结论TMEM33经RNA干扰后,能明显抑制宫颈癌的细胞生长。TMEM33可能通过影响宫颈癌细胞周期、调节细胞凋亡及细胞周期相关蛋白的表达来促进宫颈癌细胞的生长。

Objective To explore the expression of transmembrane protein 33(TMEM33)and its relationship with the prognosis in cervical cancer through bioinformatics methods,and to observe the effect of TMEM33 on the growth of cervical cancer cells after RNA interference.Methods The GEPIA(Gene Expression Profiling Interactive Analysis)database was used to select the TMEM33 genes associated with cervical cancer,and two specific siRNA sequences were designed for RNA interference experiments.The experiment was divided into 3 groups:siTMEM33#1 interference group,siTMEM33#2 interference group,and negative control(non-specific siRNA)group.TMEM33 protein expression in SiHa/HeLa cells after siRNA interference was determined by Western blot.SiHa/HeLa cells proliferation was detected by CCK-8.Apoptosis and cell cycle was detected by flow cytometry.Expression of the key genes for apoptosis BCL-2 and Bax,and the cell cycle related proteins CyclinD1 and P21 were determined by Western blot.Results Compared with siRNA negative controls,specific siRNA significantly downregulated TMEM33 protein expression,cervical cancer cell proliferation was significantly suppressed,while apoptosis was remarkably increased,cell cycle S was significantly decreased(P<0.05);key apoptosis gene BCL-2 expression was decreased while Bax expression was upregulated.CyclinD1 level was decreased while P21 level was increased(P<0.05).Conclusion After RNA interference,TMEM33 can significantly inhibit the growth of cervical cancer cells.TEME33 may promote the growth of cervical cancer cells by influencing the cervical cancer cell cycle,regulating the expression of apoptosis and cell cycle-related proteins.