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目的以替莫唑胺(temozolomide,TMZ)为阳性对照,研究新药替莫唑胺酯(temozolomide hexyl ester,TMZ-HE)在体外抑制脑胶质瘤T98G细胞生长并诱导凋亡的作用及其机制。方法 MTT比色法和克隆形成实验比较TMZ和TMZ-HE对细胞增殖的影响;流式细胞仪检测TMZ-HE和TMZ对细胞凋亡率及细胞周期的影响;Western blot检测TMZ和TMZ-HE对Bax、Bcl-2以及O~6-甲基鸟嘌呤-DNA甲基转移酶(O~6-methyl-oguanine-DNA-methyltransferase,MGMT)、γH2AX蛋白表达的影响。结果 TMZ-HE较TMZ明显抑制T98G细胞的生长,呈浓度及时间依赖性,长期作用时效果更加明显;TMZ-HE诱导T98G细胞凋亡率高于TMZ;TMZ-HE能够导致T98G细胞发生G2期阻滞;TMZ-HE能够增加Bax/Bcl-2的比值,诱导发生凋亡,TMZ-HE比TMZ更明显地消耗MGMT,产生更强的DNA损伤。结论 TMZ-HE对胶质瘤T98G细胞的增殖抑制及诱导凋亡的作用优于TMZ,消耗MGMT以及DNA损伤能力强于TMZ。
Objective To study the effect of temozolomide hexyl ester (TMZ-HE) on the growth and apoptosis of human glioma T98G cells induced by temozolomide (TMZ) as a positive control. Methods MTT assay and clonogenic assay were used to compare the effects of TMZ and TMZ-HE on cell proliferation. Flow cytometry was used to detect the effect of TMZ-HE and TMZ on the apoptosis rate and cell cycle. Western blot was used to detect TMZ and TMZ-HE On Bax, Bcl-2 and O ~ 6-methylguanine-DNA methyltransferase (MGMT), γH2AX protein expression. Results TMZ-HE significantly inhibited the growth of T98G cells in a concentration-and time-dependent manner compared with TMZ. The effects of TMZ-HE on T98G cells were more obvious than those on TMZ-HE. TMZ-HE could induce G2 phase in T98G cells TMZ-HE increased the Bax / Bcl-2 ratio and induced apoptosis. TMZ-HE consumed MGMT more significantly than TMZ, resulting in stronger DNA damage. Conclusion The effect of TMZ-HE on proliferation and apoptosis of glioma T98G cells is better than that of TMZ, MGMT depletion and DNA damage are better than TMZ.