miR-137对喉癌Hep-2细胞增殖、侵袭与凋亡的影响及可能机制

Effect and possible mechanism of microRNA-137 on the proliferation, invasion and apoptosis of laryngeal cancer Hep-2 cells

目的探讨miR-137对喉癌Hep-2细胞增殖及迁移能力的影响及可能机制。方法采用miR-137模拟物转染喉癌Hep-2细胞,实验分为空白对照组组(未进行转染)、miR-137阴性对照组(转染无关序列)、miR-137模拟物转染组(进行miR-137模拟物转染)。real-time PCR法验证转染效率,采用CCK8法检测miR-137对喉癌Hep-2细胞增殖的影响,采用Transwell实验检测转染后Hep-2细胞的侵袭能力变化,采用流式细胞术检测miR-137对Hep-2细胞凋亡的影响;Western blot检测转染后Hep-2细胞凋亡相关蛋白Bcl-2、Bax、caspase-3以及侵袭相关蛋白T细胞因子-4(TCF-4)蛋白表达的变化。结果与空白对照组或阴性对照组相比,miR-137模拟物转染组Hep-2细胞miR-137的表达水平显著高于对照组(P<0.01),喉癌Hep-2细胞的增殖与侵袭能力显著降低,细胞凋亡率明显升高,凋亡相关蛋白Bcl-2表达水平降低,而Bax与caspase-3蛋白表达升高,侵袭相关蛋白TCF-4表达降低。结论过表达miR-137抑制Hep-2细胞增殖与侵袭、诱导细胞凋亡,与上调Bax、caspase-3蛋白表达和下调Bcl-2、TCF-4表达相关。

Objective To investigate the effect of microRNA-137 on the proliferation and migration of laryngeal cancer Hep-2 cells and its possible mechanism.Methods The experiment was divided into blank control group(no transfection),miR-137 negative control group(unrelated sequence transfection),and miR-137 mimic transfection group(miR-137 mimic transfected into laryngeal cancer Hep-2 cells).Real-time PCR was used to verify the transfection efficiency,CCK8 was used to detect the effect of microRNA-137 on the proliferation of laryngeal cancer Hep-2 cells,Transwell assay was used to detect the change of invasion ability of Hep-2 cells after transfection,Flow cytometry was used to detect the effect of microRNA-137 on the apoptosis of Hep-2 cells.Western blot was used to detect the the expression of apoptosis-related proteins Bcl-2,Bax,caspase-3 and invasion-related proteins T4(TCF-4).Results Compared with blank control group or negative control group,the expression level of miR-137 was significantly higher in mimic transfection group(P<0.01),and the proliferation and invasion abilities of laryngeal cancer Hep-2 cells were significantly decreased,and the apoptotic rate was significantly increased,the expression levels of Bcl-2 and TCF-4 were decreased,but increased for the expression of Bax and caspase-3.Conclusion The inhibition of overexpressed microRNA-137 on the proliferation and invasion of Hep-2 cells is related to up-regulation of Bax,caspase-3 protein expression,down-regulation of Bcl-2 and TCF-4 expression.