miR-647通过靶基因NLRC5调控子宫内膜癌细胞的增殖和凋亡

目的:探讨miR-647对子宫内膜癌细胞增殖和凋亡的影响及其分子机制。方法:实验设置miR-NC组、miR-647组、anti-miR-NC组、anti-miR-647组、si-NC组、si-NLRC5组、miR-647+pcDNA组、miR-647+pcDNA-NLRC5组。实时荧光定量PCR（quantitative real-time PCR，qRT-PCR)检测miR-647和核苷酸寡聚化域样受体亚家族C5（nucleotide oligomerization domain-like receptor subfamily C5，NLRC5）mRNA表达水平；蛋白质印迹（Western blot）法检测NLRC5、细胞周期蛋白D1（CyclinD1）、细胞周期蛋白依赖性激酶抑制剂1A（p21）、B细胞淋巴瘤/白血病-2（Bcl-2）、Bcl-2相关X蛋白（Bax）的蛋白表达；四甲基偶氮唑盐（MTT）比色法检测细胞活性；流式细胞术检测细胞凋亡；荧光素酶报告实验检测miR-647和NLRC5的靶向关系。结果:子宫内膜癌组织中miR-647低表达，NLRC5高表达（P＜0.05）；过表达miR-647或抑制NLRC5表达，细胞活性显著降低，细胞凋亡率显著升高，CyclinD1、Bcl-2表达水平显著降低，p21、Bax表达水平显著升高（P＜0.05）。miR-647靶向调控NLRC5，NLRC5过表达逆转了miR-647过表达对子宫内膜癌Ishikawa细胞增殖和凋亡的作用。结论:过表达miR-647可能通过靶向下调NLRC5的表达抑制子宫内膜癌细胞增殖，促进细胞凋亡。

miR-647 regulates the proliferation and apoptosis of endometrial cancer cells through the target gene NLRC5

Objective:To investigate the effect of miR-647 on the proliferation and apoptosis of endometrial cancer cells and its molecular mechanism.Methods:The experiment set miR-NC group,miR-647 group,anti-miR-NC group,anti-miR-647 group,si-NC group,si-NLRC5 group,miR-647+pcDNA group,miR-647+pcDNA-NLRC5 group.Real-time fluorescence quantitative PCR（RT-qPCR) was used to detect the expression levels of miR-647 and NLRC5 mRNA.Western blot detection of nucleotide oligomerization domain-like receptor subfamily C5（NLRC5),CyclinD1,cyclin-dependent kinase inhibitor 1A（p21),B-cell lymphocytes tumor/leukemia-2（Bcl-2),Bcl-2 related X protein（Bax) protein expression.MTT assay was used to detect cell viability.Flow cytometry was used to detect apoptosis.Luciferase reporter assay was used to detect the targeting relationship between miR-647 and NLRC5.Results:Low expression of miR-647 and high expression of NLRC5 in endometrial cancer tissues（P＜0.05).Overexpression of miR-647 or inhibition of NLRC5 expression,cell viability was significantly reduces,the rate of apoptosis was significantly increases,the expressions of CyclinD1,Bcl-2 were significantly reduced,and the expressions of p21 and Bax were significantly increased（P＜0.05).miR-647 targets NLRC5,overexpression of NLRC5 reverses the effect of miR-647 overexpression on the proliferation and apoptosis of endometrial cancer Ishikawa cells.Conclusion:Overexpression of miR-647 may inhibit the proliferation of endometrial cancer cells and promote apoptosis through targeted down-regulation of NLRC5.