摘要
目的探讨抑制人滋养层细胞表面抗原-2(TROP2)基因表达对宫颈癌细胞凋亡的影响。方法空白对照组(Control组)、阴性对照无意义小干扰RNA siRNA序列组(Control siRNA组)和转染TROP2的靶向siRNA序列组(TROP2 siRNA组)转染人宫颈癌Hela细胞,48 h后蛋白印迹检测TROP2、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved caspase3)、β-连环蛋白(β-catenin)、细胞周期素D1(Cyclin D1)蛋白表达;流式细胞仪检测细胞凋亡。结果转染TROP2-siRNA后能显著降低TROP2的蛋白表达;TROP2-siRNA组细胞凋亡率及Cleaved caspase3蛋白表达显著高于Control组,β-catenin、Cyclin D1蛋白表达显著低于Control组(P<0.01),Control-siRNA组细胞凋亡率及Cleaved caspase3、β-catenin、Cyclin D1蛋白表达与Control组差异无统计学意义(P>0.05)。结论抑制宫颈癌细胞中TROP2基因表达可促进癌细胞的凋亡,其机制与上调Cleaved caspase3蛋白表达及下调Wnt/β-catenin信号通路有关。
Objective To explore the effect of inhibiting the expression of human trophoblast cell surface antigen-2（ TROP2） gene on apoptosis of cervical cancer cells. Methods The blank control group（ control group）,negative control and small interfering RNA siRNA sequence group（ control-siRNA group）,and target siRNA sequence transfected with TROP2 group（ TROP2 siRNA group） were transfected into human cervical cancer Hela cells. The expressions of TROP2,activated aspartic protein hydrolase 3 containing cysteine,β-catenin,and cyclin D1 proteins were detected after 48 hours by Western Blot. Cell apoptosis was detected by flow cytometry. Results Transfection with TROP2-siRNA could significantly reduce the expression of TROP2 protein. In TROP2-siRNA group,cell apoptosis rate and the expression level of activated aspartic protein hydrolase 3 containing cysteine were significantly higher than those in control group,the expression levels ofβ-catenin protein and Cyclin D1 protein were statistically significantly lower than those in control group（ P〈0. 01）. There was no statistically significant difference in apoptosis rate,the expression levels of activated aspartic protein hydrolase 3 containing cysteine,β-catenin,cyclin D1 proteins between control-siRNA group and control group（ P〉0. 05）. Conclusion Inhibiting the expression of TROP2 gene in cervical cancer cells can promote the apoptosis of cancer cells,the mechanism is related to upregulation of activated aspartic protein hydrolase3 containing cysteine expression and downregulation of Wnt/β-catenin signaling pathway.
出处
《中国妇幼保健》
CAS
2017年第16期3934-3937,共4页
Maternal and Child Health Care of China
