丙泊酚通过SFRP1-Wnt信号通路抑制肠癌细胞增殖侵袭的机制研究

Propofol Inhibits the Proliferation and Invasion of Colorectal Cancer Cells via Regulating SFRP1-Wnt Signaling Pathway

目的研究丙泊酚调控SFRP1-Wnt信号通路抑制肠癌细胞增殖侵袭的作用机制。方法肠癌细胞SW620及HT29分为对照组及丙泊酚组,CCK-8检测细胞增殖活性,细胞划痕实验和Transwell小室分析细胞转移和侵袭能力,逆转录实时荧光定量PCR检测Wnt信号通路相关基因,染色质免疫沉淀技术分析EZH2及H3K27Me3在SFRP1启动子区的富集程度。结果CCK-8细胞增殖试验表明肠癌细胞SW620及HT29丙泊酚处理24 h后,丙泊酚组增殖活性显著低于对照组细胞(P<0.01)。细胞划痕实验表明丙泊酚组细胞迁移愈合面积显著低于对照组(P<0.01)。Tranwell小室实验发现,丙泊酚组穿膜细胞数量显著低于对照组(P<0.01)。qPCR检测发现丙泊酚组肠癌细胞较对照细胞,E-cadherin显著上调(P<0.01),而N-cadherin表达减低(P<0.01);Wnt信号通路靶基因β-catenin、c-Myc及Cyclin D1与对照组比较明显下调(P<0.01);Wnt上游调节因子SFRP1也出现了显著表达下调(P<0.01)。染色质免疫沉淀揭示丙泊酚组细胞的EZH2、H3K27Me3在SFRP1启动子富集程度显著低于对照组细胞(P<0.01)。结论丙泊酚具有抑制肠癌细胞增殖和侵袭活性。丙泊酚通过减弱EZH2对SFRP1表观沉默作用,诱导其表达上调,从而减低Wnt信号通路活性。

Objective To investigate the effect of propofol on the proliferation and invasion of colorectal cancer cells via regulating SFRP1-Wnt signaling pathway.Methods Colorectal cancer cell lines SW620 and HT29 were used,cells were divided into 2 groups:the control group and the propofol group.CCK-8 was used to detect cell proliferation activity.The wound-healing assay and transwell assay were used to analyze cell migration and invasion ability.Reverse transcription real-time fluorescence quantitative PCR was used to detect the expression levels of Wnt signaling pathway related genes.The chromatin immunoprecipitation assay was used to analyze the enrichment of EZH2 and H3K27Me3 in the SFRP1 promoter region.Results The proliferation activity of cells in the propofol group was significantly lower than that of the control group after propofol treatment for 24 hours(P<0.01).The wound-healing assay showed that the cell migration in the propofol group was significantly lower than that in the control group(P<0.01).The tranwell assay showed that the number of transmembrane cells in the propofol group was significantly less than that in the control group(P<0.01).qPCR resutls showed that E-cadherin was significantly up-regulated(P<0.01),while N-cadherin was down-regulated(P<0.01)in the propofol group when compared with the control group.Wnt signaling pathway target genesβ-catenin,c-Myc and Cyclin D1 was significantly down-regulated when compared with the control group(P<0.01).In addition,SFRP1,the upstream regulator of Wnt,was also significantly down-regulated in the propofol group(P<0.01).Chromatin immunoprecipitation revealed that the enrichment of EZH2 and H3K27Me3 in the SFRP1 promoter was significantly lower in the propofol group than in the control group(P<0.01).Conclusion Propofol can inhibit the proliferation and invasion of colorectal cancer cells.Propofol attenuates the apparent silencing effect of EZH2 on SFRP1 and induces its upregulation,thereby reducing the activity of Wnt signaling pathway.