贵州医科大学学报

2018, v.43;No.211(04) 412-417

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雷公藤甲素对人结肠癌细胞EKR1/2、Akt、p53及AMPKα磷酸化水平的影响
Effects of Triptolide on Phosphorylation Level of EKR1/2,Akt,p53 and AMPKα

郭曼岚;漆敏;刘蔚雯;勾梦壮;孙密欣;刘亚伟;陈腾祥;
GUO Manlan;QI Ming;LIU Weiwen;GOU Mengzhuang;SUN Mixin;LIU Yawei;CHEN Tengxiang;Department of Physiology,Basic Medicine College,Guizhou Medical University;Laboratory for Precision Neurosurgery,Nanfang Hospital,Southern Medical University;First College of Clinical Medicine,Southern Medical University;The Key Laboratory of Regenerative Medicine of Guizhou Province,Guizhou Medical University;

摘要(Abstract):

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(KeyWords): 雷公藤甲素;结肠肿瘤;p53;ERK1/2;体外培养;SW480细胞
triptolide;colorectal cancer;p53;ERK1/2;culture in vitro;SW480 cells

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基金项目(Foundation): 广东省自然科学基金(2017A030313497);; 贵州省应用基础研究计划重大专项子课题[黔科合J重大字(2015)2003]

作者(Author): 郭曼岚;漆敏;刘蔚雯;勾梦壮;孙密欣;刘亚伟;陈腾祥;
GUO Manlan;QI Ming;LIU Weiwen;GOU Mengzhuang;SUN Mixin;LIU Yawei;CHEN Tengxiang;Department of Physiology,Basic Medicine College,Guizhou Medical University;Laboratory for Precision Neurosurgery,Nanfang Hospital,Southern Medical University;First College of Clinical Medicine,Southern Medical University;The Key Laboratory of Regenerative Medicine of Guizhou Province,Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2018.04.010

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