替莫唑胺耐药细胞系SHG-44/TR生物学特征及其耐药机制  

Biological characteristics and resistance mechanism of temozolomide resistance cell line SHG-44/TR

作　　者：丁凤霞[1] 朱仲玲 阎昭[1] 

机构地区：[1]天津医科大学肿瘤医院.国家肿瘤临床医学研究中心.天津市肿瘤防治重点实验室,天津300060

出　　处：《山东医药》2016年第5期19-22,共4页Shandong Medical Journal

基　　金："十二五"重大新药创制科技重大专项课题(2013ZX09303001);国家自然科学基金项目(81402481);天津卫生局科技基金项目(2014KZ085)

摘　　要：目的探讨对替莫唑胺耐药的人胶质瘤细胞系SHG-44/TR的生物学特征及耐药机制。方法通过体外分阶段剂量递增诱导法使人脑胶质瘤细胞系SHG-44对替莫唑胺产生耐药性,建立耐药细胞系SHG-44/TR。采用细胞集落形成试验和噻唑蓝比色法检测SHG-44/TR增殖情况、耐药性及耐药指数;采用Western-blotting法检测耐药蛋白O6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)及人类错配修复蛋白2(h MSH2)表达变化;流式细胞仪检测SHG-44/TR细胞周期变化。结果历时8个月成功建立了对替莫唑胺耐药的人胶质瘤细胞SHG-44/TR。与SHG-44比较,SHG-44/TR对替莫唑胺的耐受程度差异有统计学意义(P<0.05);SHG-44/TR没有细胞周期阻滞现象;SHG-44/TR较亲本细胞MGMT蛋白表达无变化,h MSH2表达差异有统计学意义(P<0.05)。结论耐替莫唑胺胶质瘤细胞系SHG-44/TR h MSH2表达降低,其耐药机制可能为DNA错配修复缺失。Objective To investigate the biological characteristics and resistance mechanism of human glioma cell line SHG-44/TR which is resistant to temozolomide（ TMZ）. Methods We induced the human glioma SHG-44 to be resistant to TMZ by dose escalation in vitro,and then established the drug-resistant cell line SHG-44/TR. The colony formation assay and MTT colorimetric assay was used to measure the proliferation,resistance and resistance index of SHG-44/TR cells; Western blotting was used to detect the expression changes of resistance protein O6-methylguanine-DNA methyl transferase（ MGMT） and human mismatch repair protein 2（ h MSH2）; and flow cytometry was used to detect the SHG-44/TR cell cycle. Results After eight months,we successfully established TMZ-resistant glioma cells SHG-44/TR. Significant difference was found in the TMZ-resistance between SHG-44 and SHG-44/TR（ P 0. 05）. SHG-44/TR had no cell cycle arrest phenomenon. The expression of MGMT protein did not change in SHG-44/TR cells as compared with that of parent cells,but the expression of h MSH2 was significantly decreased（ P 0. 05）. Conclusions The h MSH2 expression of TMZ-resistant glioma cells SHG-44/TR is decreased,and the mechanism may be DNA mismatch repair missing.

关 键 词：替莫唑胺 耐药机制 脑胶质瘤 错配修复 O6-甲基鸟嘌呤-DNA甲基转移酶 

分 类 号：R739.41[医药卫生—肿瘤]