下调miR-21表达对口腔鳞状细胞癌细胞增殖的影响

摘要

Objective: To investigate the effect of reduced expressions of miR-21 on proliferation of oral squamous cell carcinoma (OSCC) cells. Methods:Human oral cancer SCC15 and CAL27 cells were cultured. All cells were divided into miR-21 inhibitor group, miR- negative control group and blank control group, according to the different transfected. The cell proliferation in different transfected groups were tested by using MTT assay. The cell apoptosis in different transfected groups were detected. The expressions of miR-21, β-catenin, C-myc and Dkk1 were detected by using real-time quantita-tive PCR. The expressions of β-catenin, C-myc and Dkk1 proteins were detected by using Western blot. Results: After transfected in 48h and 72h, in miR-21 inhibitor group, SCC15 and CAL27 cells proliferation activities were lower than miR-negative control group and blank control group, the differences were statistically significant (P<0.05). SCC15 and CAL27 cells apoptosis rate in miR-21 inhibitor group were significantly higher than miR-negative control group and blank control groups, the differences were statistics significance (P<0.05). In miR-21 inhibitor group, the expression intensities of miR-21, β-catenin gene and protein, C-myc gene and protein were lower than miR-negative control group and blank con-trol group, while the expression intensities of Dkk1 gene and protein were higher than miR-negative control group and blank control group, the differences were all statistics significance (P<0.05). Conclusion: Down regulation of the expres-sion of miR-21 could significantly inhibit proliferation of OSCC cells and induce apoptosis. It might be related with pro-moting Dkk1 gene expression to inhibit Wnt/β-catenin signaling pathway.%目的:探讨下调miR-21表达对口腔鳞状细胞癌细胞增殖的影响及可能机制.方法:体外培养SCC15和CAL27口腔鳞癌细胞,根据转染不同,分别将细胞分为miR-21抑制物组、miR-阴性对照物组和空白对照组.MTT法检测不同转染组细胞增殖情况,检测不同转染组细胞凋亡情况,实时荧光定量PCR检测细胞中miR-21、β-catenin、C-myc和Dkk1表达,Western blot法检测细胞中β-catenin、C-myc和Dkk1蛋白表达.结果:SCC15和CAL27细胞中,miR-21抑制物组细胞转染48 h和72 h时,细胞增殖活性均低于miR-阴性对照组和空白对照组,差异均有统计学意义(P<0.05);SCC15和CAL27细胞中,miR-21抑制物组细胞凋亡率均显著高于miR-阴性对照组和空白对照组,差异均有统计学意义(P<0.05);SCC15和CAL27细胞中,miR-21抑制物组细胞中miR-21、β-catenin基因和蛋白、C-myc基因和蛋白表达量均低于miR-阴性对照组和空白对照组,而Dkk1基因和蛋白表达量均高于miR-阴性对照组和空白对照组,差异均有统计学意义(P<0.05).结论:下调miR-21可显著抑制口腔鳞状细胞癌细胞增殖,促使细胞发生凋亡,可能与促进Dkk1基因表达而抑制Wnt/β-catenin信号通路有关.