siRNA靶向抑制CXCR4基因表达对鼻咽癌细胞增殖及侵袭的影响及机制研究

The effect and mechanism of targeting inhibition of CXCR4 gene expression by siRNA on proliferation and invasion of nasopharyngeal carcinoma cells

目的探讨抑制鼻咽癌细胞趋化因子受体-4(CXCR4)基因表达对癌细胞增殖、侵袭的影响及机制。方法回顾性收集鼻咽癌42例临床资料及相应的癌旁组织。RT-PCR和Western bloting分别检测鼻咽癌及相应的癌旁组织中CXCR4基因的mRNA及蛋白表达;人鼻咽癌细胞系CNE-2Z分为空白组(细胞未做任何处理)、NC-siRNA组(细胞转染无义siRNA序列)和CXCR4-siRNA组(细胞中转染CXCR4的靶向siRNA序列)。转染48 h后Western bloting检测各组细胞中CXCR4、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、β-连环蛋白(β-catenin)、细胞周期素D1(Cyclin D1)蛋白表达;CCK8实验和Transwell小室分别检测细胞的增殖及侵袭能力。结果鼻咽癌中CXCR4基因的mRNA及蛋白表达均显著高于癌旁组织(P<0.01);转染CXCR4-siRNA组后CXCR4的蛋白表达显著降低;NC-siRNA组细胞存活率、细胞侵袭数及MMP-2、MMP-9、β-catenin、Cyclin D1蛋白表达与空白组差异无统计学意义(P>0.05);CXCR4-siRNA组细胞存活率、细胞侵袭数及MMP-2、MMP-9、β-catenin、Cyclin D1蛋白表达均显著低于空白组(P<0.01)。结论抑制鼻咽癌细胞CXCR4基因表达可显著降低癌细胞的增殖与侵袭能力,其机制与下调Wnt/β-catenin信号通路有关。

Objective To investigate the effect and mechanism of targeting inhibition of CXCR4 gene expression by siRNA on proliferation and invasion of nasopharyngeal carcinoma cells. Methods 42 cases of nasopharyngeal carcinoma and corresponding paracancerous tissues were retrospectively collected. RT-PCR and Western bloting were used to detect mRNA and protein expression of CXCR4 gene in nasopharyngeal carcinoma and corresponding adjacent tissues; human nasopharyngeal carcinoma cell line CNE-2 Z were divided into Control group,NC-siRNA group and CXCR4-siRNA group,protein expression of CXCR4,MMP-2,MMP-9,β-catenin,Cyclin D1 after transfected for 48 h were detected by Western bloting; CCK8 experiment and Transwell chamber were used to detect the proliferation and invasion of cells. Results The mRNA and protein expression of CXCR4 gene in human nasopharyngeal carcinoma were significantly higher than those in adjacent tissues（ P〈0. 01）.The expression of CXCR4 protein after transfected CXCR4-siRNA was significantly lower. The cell survival rate,cell invasion and MMP-2,MMP-9,β-catenin,Cyclin D1 protein expression in NC-siRNA group had no significant difference with Control group（ P〈0. 05）. The cell survival rate,cell invasion and MMP-2,MMP-9,β-catenin,Cyclin D1 protein expression in CXCR4-siRNA group were significantly lower than Control group（ P〈0. 01）. Conclusion Inhibiting of CXCR4 gene expression in nasopharyngeal carcinoma cells can significantly decrease the proliferation and invasion ability of cancer cells,and the mechanism is related to down regulation of Wnt/β-catenin signaling pathway.