【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。更多还原

【Abstract】 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 μM,5 μM) and TMZ( 0. 5 m M,1. 5 m M,5 m M) 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.更多还原