TROP2基因表达对食管癌细胞增殖及侵袭能力的影响

Effects of TROP2 gene expression on proliferation and invasion of esophageal cancer cells

目的探讨抑制食管癌细胞人滋养层细胞表面抗原-2(human trophoblast cell-surface antigen 2,TROP2)基因表达对肿瘤细胞增殖及侵袭的影响及机制。方法 TROP2-siRNA转染人食管癌EC9706细胞(TROP2-siRNA组),同时设置对照组(仅加入脂质体)和阴性对照NC-siRNA组(加入合成的无干扰作用的siRNA)。转染48 h后,Western blot检测各组细胞中TROP2、Ki-67、基质金属蛋白2(matrix metalloproteinase 2,MMP-2)、MMP-9、磷脂酰肌醇-3激酶(phosphoinositide 3-kinase,PI3K)和磷酸化的丝氨酸苏氨酸激酶(phosphorylated serine/threonine kinase,p-AKT)的蛋白表达,CCK8实验检测细胞增殖,transwell小室检测细胞侵袭能力。结果转染TROP2-siRNA后EC9706细胞中TROP2蛋白表达降低(P<0.05)。NC-siRNA组细胞存活率、细胞侵袭数及Ki-67、MMP-2、MMP-9、PI3K、p-AKT蛋白表达与对照组比较,差异均无统计学意义(均P>0.05),而TROP2-siRNA组均低于对照组与NCsiRNA组(均P<0.05)。结论抑制食管癌TROP2基因表达可降低食管癌细胞的增殖及侵袭能力,其机制与下调Ki-67、MMP-2、MMP-9表达及PI3K/AKT信号通路有关。

Objective To investigate the effects and the mechanism of human trophoblast cell-surface antigen 2（ TROP2） gene expression on the proliferation and invasion of esophageal cancer cells. Methods Human esophageal carcinoma EC9706 cells were transfected with TROP2-siRNA（ TROP2-siRNA group）,liposome（ normal control group）,or non-interference siRNA（ NC-siRNA group）,respectively. Western blot was used to examine the levels of TROP2,Ki-67,matrix metalloproteinase 2（ MMP-2）,MMP-9,phosphoinositide 3-kinase（ PI3 K） and phosphorylated serine/threonine kinase（ p-AKT） protein at 48 h after transfection. CCK8 assay was used to analyze cell proliferation,and transwell assay was used to detect cell invasion ability. Results After TROP2-siRNA transfection,the expression of TROP2 protein was significantly decreased in EC9706 cells（ P〈0. 05）. Moreover,the cell survival rates,cell invasion and the expression of Ki-67,MMP-2,MMP-9,PI3 K,p-AKT proteins were all significantly decreased in the TROP2-siRNA group as compared with the normal control group and NC-siRNA group（ all P〈0. 05）. Conclusion TROP2 gene expression is involved in the proliferation and invasion ability of esophageal cancer cells,which is associated with Ki-67,MMP-2,MMP-9 expression and the PI3 K/AKT signaling pathway.