新药替莫唑胺酯抗原发性耐药胶质瘤机制研究

Mechanism of New Drug Temozolomide Easter on Primary Resistance Glioma

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 investigate the effect of temozolomide（TMZ） hexyl easter（TMZ-HE） on the proliferation and apoptosis of brain glioma T98 G cells in vitro compared with TMZ, and then illustrate the mechanism. METHODS Methyl thiazolyl tetrazolium（MTT） assay and colony formation assay was used to compare the effect of TMZ and TMZ-HE on cell proliferation. Cell apoptosis rate and cell cycle induced by TMZ and TMZ-HE were analyzed by flow cytometry. Western blot was used to detect the expression of Bax, Bcl-2, O^6-methyl-oguanine-DNA-methyltransferase（MGMT）, γH2AX after treated with TMZ and TMZ-HE. RESULTS TMZ-HE obviously suppressed the cell proliferation in a concentration and time-dependent manner compared with TMZ, and it was more obvious in long-term effect; TMZ-HE induced more cellular apoptosis of the T98 G cells than TMZ; TMZ-HE induced a distinct G2 arrest in T98 G cell; TMZ-HE induced apoptosis by increasing the ratio of Bax/Bcl-2, TMZ-HE consumed MGMT more significant than TMZ, and produced more DNA damage. CONCLUSION TMZ-HE has an advantage over the effect of anti-proliferative and induced apoptosis than TMZ, and the ability of TMZ-HE to consume MGMT and DNA damage is improved.