miR-196a/LRIG3在结直肠癌组织中的表达及对HCT116细胞增殖、侵袭的影响

Expression of miR-196a/LRIG3 in Colorectal Cancer Tissues and Its Effects on Proliferation and Invasion of HCT116 Cells

目的探讨miR-196a/免疫球蛋白样结构域3(LRIG3)在结直肠癌(CRC)组织中表达及对CRC HCT116细胞增殖、侵袭的影响。方法检测CRC患者癌组织和癌旁正常组织中miR-196a和LRIG3 mRNA的表达,比较不同临床特征患者癌组织中miR-196a和LRIG3 mRNA表达差异,并采用Pearson相关分析探讨其相关性。用Lipofectamine法将miR-196a inhibitor和miR-NC转染到HCT116细胞,48 h后分别采用MTT法和Transwell法检测细胞增殖率及细胞侵袭能力,同时应用Western blot法检测HCT116细胞中LRIG3、E-钙黏蛋白(E-cadherin)和N-钙黏蛋白(N-cadherin)表达。结果癌组织miR-196a mRNA相对表达量高于癌旁正常组织(P<0.05);癌组织LRIG3 mRNA相对表达量低于癌旁正常组织(P<0.05);癌组织中miR-196a和LRIG3 mRNA水平与TNM分期、肿瘤分化程度、肿瘤浸润深度和肿瘤转移相关(P<0.05,P<0.01)。Pearson相关分析显示,miR-196a和LRIG3 mRNA在CRC组织中的表达呈负相关(r=-0.442,P=0.013)。与空白对照组比较,miR-196a inhibitor组HCT116细胞增殖率和侵袭细胞数降低,LRIG3和N-cadherin蛋白表达增加,E-cadherin蛋白表达降低(P<0.05);与miR-196a inhibitor组比较,miR-NC组HCT116细胞增殖率和侵袭细胞数增加,LRIG3和N-cadherin蛋白表达降低,E-cadherin蛋白表达增加(P<0.05)。结论CRC组织中miR-196a表达升高,LRIG3表达降低,二者在CRC组织中的表达呈负相关,miR-196a可通过靶向作用上调LRIG3表达从而抑制HCT116细胞增殖和侵袭。

Objective To investigate the expression of miR-196a/LRIG3 in colorectal cancer(CRC)tissue and its effects on proliferation and invasion of CRC HCT116 cells.Methods Expressions of miR-196a and LRIG3 mRNA in cancer tissues and adjacent normal tissues of CRC patients were detected,and the differences of miR-196a and LRIG3 mRNA expressions in cancer tissues of patients with different clinical characteristics were compared,and their correlations were analyzed by Pearson correlation analysis.The miR-196a inhibitor and miR-NC were transfected into HCT116 cells by Lipofectamine method.After transfection for 48 h,cell proliferation rate and cell invasion ability were detected by MTT method and Transwell method respectively.In the meantime,expressions of LRIG3,E-cadherin and N-cadherin proteins in HCT116 cells were detected by Western blot.Results The relative expression of miR-196a mRNA in cancer tissues was significantly higher than that in adjacent normal tissues(P<0.05),while the relative expression of LRIG3 mRNA in cancer tissues was significantly lower than that in adjacent tissues(P<0.05).Levels of miR-196a and LRIG3 mRNA were correlated with TNM stage,degree of tumor differentiation,depth of tumor invasion and tumor metastasis in cancer tissues(P<0.05,P<0.01).Pearson correlation analysis showed that there was a negative correlation between the expressions of miR-196a with LRIG3 mRNA in CRC tissues(r=-0.442,P=0.013).Compared with those in blank control group,proliferation rate and number of invasive cells of HCT116 cells were significantly decreased,while expressions of LRIG3 and N-cadherin proteins were significantly increased,and the expression of E-cadherin protein was significantly decreased in miR-196a inhibitor group(P<0.05).Compared with those in miR-196a inhibitor group,proliferation rate and number of invasive cells of HCT116 cells were significantly increased,while expressions of LRIG3 and N-cadherin proteins were significantly decreased,and the expression of E-cadherin protein was significantly increased in miR-NC group(P<0.05).Conclusion The expression of miR-196a is increased,and LRIG3 expression is decreased in CRC tissues.There is a negative correlation between expressions of miR-196a with LRIG3 in CRC tissues.The miR-196a may inhibit proliferation and invasion of HCT116 cells by targeting up-regulation of LRIG3 expression.