作　　者： ; ; ;

机构地区： 贵州医科大学基础医学院生理学教研室

出　　处： 《贵州医科大学学报》 2018年第4期412-417,共6页

摘　　要： 目的:观察雷公藤甲素(Tp)对人结肠癌细胞EKR1/2、Akt、p53及AMPKα等信号通路分子的影响。方法:体外培养人结肠癌SW480细胞,用40 nmol/L Tp刺激24 h,分别于刺激0、24 h时对细胞进行拍照,观察细胞形态,应用Path Scan分别检测细胞内信号通路分子ERK1/2的Thr202/Tyr204位点、Akt的Thr308和Ser473位点,AMPKα的Thr172位点及p53的Ser15位点的磷酸化水平,并通过Western blot验证p53蛋白在人结肠癌细胞SW480,SW620和HCT116细胞中的磷酸化水平。结果:40 nmol/L Tp刺激24 h后,人大肠癌SW480细胞变大、形态不规整、触角变长,细胞表型发生改变;Path Scan结果表明,Tp刺激24 h时,SW480细胞ERK1/2的Thr202/Tyr204位点、Akt的Thr308和Ser473位点及p53的Ser15位点磷酸化水平增高,AMPKα的Thr172磷酸化水平降低;Western blot实验结果显示,Tp刺激24 h时,人结肠癌SW480、SW620和HCT116细胞中p53蛋白的磷酸化水平上调。结论:Tp可能是通过ERK1/2、Akt、p53和AMPKα信号通路影响结肠癌细胞表型改变。 Objective： To observe the effects of triptolide（ Tp） on the phosphorylation of intracellular signaling pathway molecules in human colorectal cancer cells EKR1/2,Akt,p53,and AMPKα.Methods： Human colorectal cancer SW480 cells were cultured in vitro and stimulated by 40 nmol/L Tp for 24 h. Path Scan assay was used to detect the phosphorylation of intracellular signaling pathway molecules in ERK1/2 at Thr202/Tyr204,Akt at Thr308/Ser473,AMPKα at Thr172 and p53 at Ser15. The phosphorylation of p53（ Ser15） in SW480,SW620 and HCT116 cells were verified by Western blot. Results： The cellular morphology during the treatment with 40 nmol/L Tp for 24 h observed by the microscope showed significant changes. Path Scan method detected some important molecules in ERK1/2,Akt,p53 and AMPKα which were activated by phosphorylation within the intracellular signaling pathways. Western blot demonstrated that the level of p-p53（ Ser15） was up-regulated in SW480,SW620 and HCT116 cells of human colorectal cancer. Conclusions： Triptolide,Tp can change the phenotype of colorectal cancer cells,which is likely to be developed through activation of ERK1/2,Akt,p53,and AMPKα in signaling pathways.

关 键 词： 雷公藤甲素 结肠肿瘤 体外培养 细胞

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