替莫唑胺诱导的耐药细胞系U251/TR的构建及衣霉素对其耐药性的逆转作用  

Establishment of drug-resistance U251/TR cell line induced by temozolomide and reverse effects of tunicamycin

作　　者：杨艳茹[1] 刘景景[1] 孙利利[1] 赵晶[1] 王浩[1] 马健[1] 王德才[1] 

机构地区：[1]泰山医学院药学院,山东泰安271016

出　　处：《泰山医学院学报》2016年第7期721-723,共3页Journal of Taishan Medical College

基　　金：山东省自然科学基金(ZR2011HQ055);国家自然科学基金(81302202;81441111)

摘　　要：目的构建替莫唑胺（temozolomide,TMZ）耐药细胞系U251/TR,并观察衣霉素（tunicsmycin,Tm）逆转对替莫唑胺耐药细胞系U251/TR细胞的作用。方法采用替莫唑胺对U251细胞进行体外分步诱导,产生耐药的U251/TR细胞系;CCK-8法检测TMZ对U251和U251/TR细胞增殖抑制作用,计算半数抑制浓度（IC50）,得到U251/TR细胞系对TMZ的耐药指数（RF）;采用细胞集落法确定U251/TR的耐药性;将Tm联合TMZ作用于U251/TR细胞系,采用CCK-8法分别测定不同给药组细胞的增殖抑制作用。结果通过体外分步诱导法成功地建立了对TMZ具有稳定耐药性的U251/TR细胞株。CCK-8法检测结果显示,U251/TR的耐药指数约为5.2;细胞集落法确定诱导过程中的U251/TR的耐药性为1.2-3倍。Tm和TMZ联合应用呈协同效应。结论成功建立了由TMZ诱导的人脑胶质瘤耐药细胞株U251/TR。Tm联合TMZ能显著逆转U251/TR细胞系的耐药性。Objective： To establish drug-resistance U251/TR cell line with timozolomide（TMZ）, and to study the effect of tunicamycin （Tm） on reversing temozolomide resistance in glioma cells. Methods： Drug resistance of U251 glioma cells was induced by the stepwise revulsion with TMZ. The proliferative inhibition effect of TMZ on U251, U251/TR cell line and IC50 value were evaluated by CCK-8 method. The drug resistance and resistance index （RI） were determined by cell colony experiment and CCK-8 assay. The combination of Tm （ 1 p,M, 5 p,M ） and TMZ （ 0. 5 raM, 1.5 mM, 5 raM） on U251/TR cells were tested by CCK-8 method. Results： U251/TR cell line of drug resistance was successfully induced by TMZ with step wise revulsion method in 6-month culture. The cell cloning experiment demonstrated that the colony-foming efficency of U251/TR cells was 1.2 - 3 times that of U251 cells. The drug resistance index was about 5.2 in U251/TR cell line assayed by CCK-8 method. Combined Tm and TMZ treatment showed a synergistic effect in U251/TR cell （Q 〉 1.15 ）. Conclusion： The drug resistant U251/TR cell line was established successfully. Combination of Tm with TMZ could reverse durg resistance of U251/TR cells.

关 键 词：替莫唑胺 脑胶质瘤 耐药性 衣霉素 

分 类 号：R96[医药卫生—药理学]