siRNA干扰Cx43表达抑制膀胱癌细胞侵袭的机制研究

Mechanism of siRNA interference Cx43 expression to inhibit the invasion of bladder cancer cells

目的探讨siRNA干扰Cx43表达抑制膀胱癌细胞侵袭的机制。方法将膀胱癌细胞株5637随机分为A、B、C组。A组未进行任何转染,B组转染空白质粒,C组转染Cx43-siRNA表达载体。转染后,以Transwell法检测细胞侵袭能力,以逆转录聚合酶链式反应(RT-PCR)法检测Cx43 mRNA表达,以蛋白质印迹(Western blot)法检测Cx43、P38、基质金属蛋白酶(MMP)-2、MMP-9表达。结果转染72h后,A、B、C组Cx43 mRNA相对表达量分别为1.00±0.06,0.96±0.08,0.32±0.07,Cx43蛋白相对表达量分别为1.02±0.07,0.99±0.08,0.27±0.06,MMP-2蛋白表达水平分别为0.93±0.07,0.96±0.04和0.25±0.07,MMP-9蛋白表达水平分别为1.07±0.08,1.02±0.05和0.18±0.03,P38蛋白表达水平分别为1.47±0.25,1.32±0.18和0.37±0.08,穿膜细胞数分别为2.05±0.08,2.13±0.14和0.48±0.06。B组与A组比较,上述指标水平差异均有统计学意义(均P<0.05);C组与B组相比较,上述指标水平差异均有统计学意义(均P<0.05)。结论Cx43表达与膀胱癌细胞的侵袭呈正相关,siRNA干扰Cx43表达可降低膀胱癌的侵袭能力,对进展期膀胱癌的治疗具有潜在前景。

Objective To investigate the mechanism of siRNA interference Cx43 expression inhibiting the invasion of bladder cancer cells.Methods Bladder cancer cell line 5637 was randomly divided into group A,B and C.No transfection was carried out in group A,blank plasmid was transfected in group B,and Cx43-siRNA expression vector was transfected in group C.After transfection,the invasive ability of cells was detected by Transwell test,the expression of Cx43 mRNA was detected by reserve transcription polymerase chain reaction(RT-PCR),and the expression of Cx43,P38,MMP-2 and MMP-9 was detected by Western blot.Results At 72 h after transfection,the relative expression of Cx43 mRNA in group A,B and C were 1.00±0.06,0.96±0.08,0.32±0.07,the relative expression of Cx43 protein were 1.02±0.07,0.99±0.08 and 0.27±0.06,the expression of MMP-2 protein were 0.93±0.07,0.96±0.04 and 0.25±0.07,the expression of MMP-9 protein were 1.07±0.08,1.02±0.05 and 0.18±0.03,the expression levels of P38 protein were 1.47±0.25,1.32±0.18 and 0.37±0.08,the number of perforating cells were 2.05±0.08,2.13±0.14 and 0.48±0.06.There were significant differences in above indexes between group B and group A(all P<0.05).There were significant differences in above indexes between group C and group B(all P<0.05).Conclusion The expression of Cx43 is positively correlated with the invasion of bladder cancer cells.Interference of Cx43 expression by siRNA can reduce the invasive ability of bladder cancer and has potential prospects for the treatment of advanced bladder cancer.