siRNA沉默MUC16对胆囊癌细胞增殖和迁移的影响

Effect of siRNA silencing MUC16 on the proliferation and migration of gallbladder carcinoma cells

目的:探讨siRNA沉默黏蛋白(mucin,MUC)16基因对人胆囊癌细胞(GBC-SD)增殖和迁移的影响及其分子机制。方法:采用慢病毒感染和siRNA干扰技术沉默GBC-SD中MUC16基因(GBC-SD+MUC16-siRNA组),同时设立阴性对照组(GBC-SD+NC组)和空白对照组(GBC-SD组)。RT-PCR检测各组GBC-SD细胞中MUC16表达情况,噻唑蓝实验检测GBC-SD细胞增殖能力,划痕实验检测GBC-SD细胞迁移能力,蛋白免疫印迹实验检测GBC-SD细胞中PCNA和CD44蛋白表达情况。结果:GBC-SD+MUC16-siRNA组细胞增殖率和细胞迁移数目均低于GBC-SD组(P<0.05);GBC-SD+MUC16-siRNA组MUC16、CD44蛋白表达均低于GBC-SD组、GBC-SD+NC组(P<0.05~P<0.01),PCNA蛋白表达亦低于GBC-SD+NC组(P<0.05)。结论:siRNA沉默MUC16具有一定的抑制胆囊癌细胞增殖和迁移作用,可为胆囊癌的治疗提供新的实验证据。

Objective:To explore the effects of siRNA silencing mucin 16(MUC16)on the proliferation and migration of gallbladder carcinoma GBC-SD cells,and its molecular mechanisms.Methods:The GBC-SD+MUC16-siRNA group was constructed using the lentivirus infection and siRNA interference technique to silence MUC16 gene in GBC-SD cells,and the negative control group(GBC-SD+NC group)and blank control group(GBC-SD group)were set up simultaneously.The RT-PCR was used to detect the expression of MUC16 in GBC-SD cells,the MTT and scratch assays were used to detect the proliferation and migration ability of GBC-SD cells,and the Western blotting was used to detect the protein expressions of CD44 and PCNA in GBC-SD cells.Results:The cell proliferation rate and cell migration number in GBC-SD+MUC16-SiRNA group were lower than those in GBC-SD group(P<0.05).The expression levels of MUC16 and CD44 protein in GBC-SD+MUC16-SIRNA group were lower than those in GBC-SD group and GBC-SD+NC group(P<0.05 to P<0.01),and the expression level of PCNA protein in GBC-SD+MUC16-SIRNA group was also lower than that in GBC-SD+NC group(P<0.05).Conclusions:SiRNA silencing MUC16 can inhibit the proliferation and migration of gallbladder cancer cells,which may provide the new experimental evidence for the treatment of gallbladder cancer.