TET1介导5-羟甲基胞嘧啶表达抑制突变型IDH1过表达胶质瘤细胞增殖、迁移和侵袭  

Effects of 5-hydroxymethylcytosine on biological behavior of IDH mutant glioma

作　　者：雷梓[1] 陈建功 何颖 李睿[1] 吴莹莹[1] LEI Zi;CHENG Jiangong;HE Ying;LI Rui;WU Yingying(Department of pathology,The First Affiliated Hospital of Kunming Medical University,Kunming,Yunnan,China,650032;Department of pathology,Guangnan People's Hospital,Wenshan,Yunnan,663300)

机构地区：[1]昆明医科大学第一附属医院病理科,云南昆明650032 [2]广南县人民医院,云南文山663300

出　　处：《分子诊断与治疗杂志》2021年第4期582-585,589,共5页Journal of Molecular Diagnostics and Therapy

基　　金：云南省教育厅科学研究基金项目(2018JS210);云南省科技厅省基础研究计划(昆医联合专项)[2019FEO01(-211)];国家自然科学基金(8166010413)。

摘　　要：目的探讨TET1介导的5-羟甲基胞嘧啶(5hmC)表达对异柠檬酸脱氢酶1(IDH1)突变的U251胶质瘤细胞生物学行为的影响。方法构建R132H突变型IDH1(IDH1^(R132H))过表达载体,转染U251人胶质瘤细胞系,实时荧光定量PCR(RT-qPCR)及蛋白质免疫印迹(WB)检测IDH1^(R132H)及TET1表达水平,免疫荧光检测5hmC表达水平。IDH1^(R132H)过表达载体与TET过表达载体共转染U251细胞,RT-qPCR和WB检测TET1表达水平,利用CCK8实验检测肿瘤细胞增殖能力,划痕实验检测肿瘤细胞迁移能力,transwell小室实验检测肿瘤细胞侵袭能力,流式细胞实验检测肿瘤细胞周期及细胞凋亡。结果在IDH1^(R132H)过表达U251细胞中,TET1及5hmC表达水平降低,差异有统计学意义(P<0.05)。IDH1^(R132H)过表达U251细胞中过表达TET1后,5hmC水平升高,差异有统计学意义(P<0.05),同时抑制肿瘤细胞的增殖、迁移、侵袭能力,促进肿瘤细胞凋亡(P<0.05)。结论 TET1介导的5hmC表达能抑制IDH1^(R132H)过表达U251胶质瘤细胞的增殖、侵袭、迁移能力,促进肿瘤细胞凋亡。Objective To investigate the effect of TEF1-mediated 5-hydroxymethylcytosine(5 hmC)expression on the biological behavior of U251 glioma cells with isocitrate dehydrogenase 1(IDH1)mutation.Methods The R132H mutant IDH1(IDH1^(R132H))overexpression vector was constructed,transfected into the U251 human glioma cell line,and the expression levels of IDH1^(R132H)and TET1 were detected by real-time fluorescent quantitative PCR(RT-qPCR)and Western blotting(Western Blotting,WB).5 hmc expression level was detected by immunofl uorescence.IDH1^(R132H)overexpression vector and TET1 overexpression vector were cotransfected into U251 cells,TET1 expression level was detected by RT-qPCR and WB,tumor cell proliferation ability was detected by CCK8 experiment,tumor cell migration ability was detected by scratch test,and tumor cell invasion ability was detected by transwell chamber experiment Ability,flow cytometry test to detect tumor cell cycle and apoptosis were deteced by flow cytometry.Results In IDH1^(R132H)overexpressing U251 cells,the expression levels of TET1 and 5 hmC decreased,and after overexpressing TET1 in IDH1^(R132H)overexpressing U251 cells,the 5 hmC level increased,inhibiting the proliferation,migration and invasion of tumor cells,and promoting tumor cell apoptosis.Conclusion TET1-mediated 5 hmC expression can inhibit the proliferation,invasion and migration of U251 glioma cells overexpressing IDH1^(R132H),and promote tumor cell apoptosis.

关 键 词：IDH突变 U251胶质瘤细胞 TET1 5hmC 

分 类 号：R739.41[医药卫生—肿瘤]