丙泊酚减低EZH2对miR-34a抑制效应与肠癌细胞增殖侵袭能力变化的研究

Relationship between propofol reducing inhibitory effect of EZH2 on miR-34a and proliferation,invasion of colorectal cancer cells

目的:研究丙泊酚对EZH2-miR34a调控影响与对肠癌细胞增殖侵袭作用的关系。方法:实时荧光定量PCR检测miR-34a、EZH2表达。染色质免疫沉淀技术检测EZH2在miR-34a启动子区富集程度、CCK-8分析细胞增殖活性变化、Transwell小室试验检测细胞侵袭能力变化。结果:肠癌细胞SW480、HCT116丙泊酚组与对照组比较,miR-34a显著上调(P<0.01),而EZH2表达水平明显抑制(P<0.01)。染色质免疫沉淀技术表明丙泊酚组细胞EZH2在miR-34a启动子富集程度显著低于对照组细胞(P<0.01)。CCK-8显示与对照相比,丙泊酚和miR-34a mimics处理组SW480、HCT116细胞48 h后,显著抑制其增殖活性(均P<0.01);丙泊酚+miR-34a inhibitor联合处理组,细胞增殖活性与对照组比较无显著变化(P>0.05)。Transwell小室试验表明丙泊酚和miR-34a mimics处理组细胞与对照组比较,穿膜细胞数显著减少(P<0.01)。丙泊酚与miR-34a inhibitor联合处理组与对照组比较,穿膜细胞数无显著变化(P>0.05)。结论:丙泊酚对肠癌细胞的增殖和侵袭能力具有显著抑制能力。丙泊酚通过减低EZH2的表达及在miR-34a启动子的富集,促进miR-34a转录表达。

Objective:To investigate the relationship between the effect of propofol on the regulation of EZH2-miR34 a and the proliferation,invasion of colorectal cancer cells.Methods:Real-time fluorescence quantitative PCR was used to detect the expression of miR-34 a and EZH2.Chromatin immunoprecipitation was used to detect the enrichment of EZH2 in the miR-34 a promoter region.CCK-8 was used to analyze the changes of cell proliferation activity.Transwell assay was used to detect the changes of cell invasion ability.Results:Compared with the control group,the miR-34 a of colorectal cancer cells SW480 and HCT116 in propofol group were significantly up-regulated,while the expression level of EZH2 was significantly inhibited(P<0.01).Chromatin immunoprecipitation assay showed that the enrichment degree of EZH2 in the miR-34 a promoter in propofol group was significantly lower than that in control group(P<0.01).CCK-8 showed that,compared with the control,the propofol and miR-34 a mimics treated SW480 and HCT116 cells significantly inhibited their proliferative activity after 48 hours(all P<0.01),but there was no significant change in proliferation activity between the propofol combined with miR-34 a inhibitor group and the control group(P>0.05).Transwell test showed that,compared with the control group,the number of transmembrane cells in the propofol and miR-34 a mimics treatment groups was significantly decreased(P<0.01),but there was no significant change in the number of transmembrane cells between the propofol combined with miR-34 a inhibitor group and the control group(P>0.05).Conclusion:Propofol can significantly inhibit the proliferation and invasion of colorectal cancer cells.Propofol promotes miR-34 a transcriptional expression by reducing EZH2 expression and enrichment at the miR-34 a promoter.