二甲双胍通过调控TET2对子宫内膜癌细胞增殖的影响

The effect of metformin on the proliferation of endometrial cancer cells by regulating TET2

目的探究二甲双胍对子宫内膜细胞增殖影响及调控机制。方法用二甲双胍(Met)药物处理子宫内膜癌细胞ishikawa和HEC-1A,设置药物浓度(0 mmol/L、1 mmol/L、5 mmol/L、10 mmol/L、15 mmol/L)并对EC细胞加药处理,检测细胞增殖速度变化,细胞周期分布和细胞凋亡百分比,细胞TET2在蛋白表达和转录水平变化,实时定量PCR检测5-羟甲基胞嘧啶(5-hmC)在细胞中含量变化,检测子宫内膜癌组织芯片中TET2和5-hmC蛋白表达,以ishikawa细胞为研究对象,功能学实验检测过表达TET2蛋白后细胞增殖变化。结果EC细胞经二甲双胍处理细胞增殖能力显著降低(P<0.01),经10mmol/L二甲双胍作用EC细胞G0/G1和S期的DNA含量以及凋亡细胞的百分比显著增加(P<0.05),经二甲双胍药物处理48 h后TET2蛋白和TET2 mRNA表达显著上调,并且两组细胞中5-hmC DNA含量水平升高(P<0.05);TET2蛋白和5-hmC在组织中显著低表达(P<0.01),外源性升高TET2蛋白表达EC细胞增殖趋势和经二甲双胍处理后的趋势一致。结论二甲双胍通过TET2/5-hmC信号通路抑制细胞增殖。

Objective To explore the effects of metformin on endometrial cell proliferation and its regulatory mechanism.Methods With metformin(Met)drug treatment of endometrial cancer cells ishikawa and HEC-1A,set up the drug concentrations(0 mmol/L,1 mmol/L,5 mmol/L,10 mmol/L,15 mmol/L)and the EC cells dosing process,detect changes in the rate of cell proliferation,Cell cycle distribution and percentage of apoptosis,changes in protein expression and transcription level of TET2 cells,real-time quantitative PCR detection of 5-hydroxymethyl cytosine(5-hmC)content changes in cells,detection of TET2 and 5-hmC protein expression in endometrial cancer tissue chip,ishikawa cells as the research object.Results cell proliferation after overexpression of TET2 protein were detected by functional experiments:The proliferation ability of EC cells treated with metformin was significantly decreased(P<0.01),and DNA content of G0/G1 and S phase and percentage of apoptotic cells were significantly increased after 10 mmol/L metformin treatment(P<0.05).After treated with metformin for 48 h,TET2 protein and TET2 protein were significantly increased MRNA expression was significantly up-regulated,and 5-hmC DNA content was increased in both groups(P<0.05).The expression of TET2 protein and 5-hmC was significantly lower in tissues(P<0.01),and the proliferation trend of EC cells with exogenous elevated TET2 protein expression was consistent with that of metformin treated EC cells.Conclusion Metformin inhibits cell proliferation through TET2/5-hmC signaling pathway.