卡氮芥联合全反式维甲酸诱导脑胶质瘤的凋亡及其分子机制

BCNU combined with all-trans retinoic acid induces glioma apoptosis and mecbanism

目的探讨全卡氮芥（BCNU）联合全反式维甲酸（ATRA）诱导脑胶质瘤的凋亡及其分子机制。方法分为空白对照组、ATRA组、BCNU组、BCNU＋ATRA组，噻唑蓝（MTT）比色法计算生长抑制率。FCM检测药物作用24h后细胞周期和凋亡率变化。逆转录一聚合酶链反应（RT—PCR）法检测CyclinE、CDK2、p27Kipl mRNA表达变化。结果与联合用药组比较C6、U251细胞抑制率分别为（10．32±1．12）％至（40．06±1．52）％、（6．39±0．65）％至（35．30±1．67）％；C6、U251细胞G．期比例分别由（42．18±2．52）％至（63．04±4．05）％、（57．144-2．52）％至（69．50±4．05）％，C6、U251细胞凋亡率2．46±1．02至17．43±2．03、10．36±1．76至21．32±2．03；Cyelin E、CDK2mRNA表达下调，p27Kipl mRNA表达上调。结论BCNU与ATRA协同应用诱导脑胶质瘤细胞的凋亡，其诱导凋亡的分子机制可能是通过改变相关细胞周期蛋白导致细胞周期改变。

Objective To explore the mechanism that BCNU combined with ATRA induces glioma apoptosis. Methods MTT assay was used to observe the effect of control,ATRA .BCNU and combined group on the growth of C6,U251 cell lines. Flow cytometry was used to determine the cell cycle distribution and the rate of apoptosis in 24h. RT-PCR was used to detect expression of Cyclin E, CDK2, p27Kipl mRNA. Results MTT showed that the inhibition rate varied from （ 10.32 ± 1.12） % to （40.06 ± 1.52 ） %, （6.39 ± 0.65 ） % to （ 35.30 ± 1.67 ） % in C6 and U251 cells respectively compared with compared drugs. In flow cytometry analysis glioma cells arrested in G1 phase from （42.18 ±2.52）% to （63.04 ± 4.05 ） %, （ 57.14 ± 2.52 ） % to （ 69.50 ± 4.05 ） % and the rate of apoptosis was increased from （2.46 ± 1.02） to （17.43 ±2.03） ,（10.36 ± 1.76） to （21.32 ±2.03） in C6 and U251 cells respectively. RT-PCR showed combined drugs group can obviously down-regulate the expression of Cyclin E, CDK2 mRNA and up-regulate the expression of p27Kipl mRNA in contrast to single medicine and control group （ P 〈 0.05）. Conclusion BCNU combined with ATRA can induce C6, U251 glioma cells apoptosis. The mechanism may be to alter relevant cell cycle protein.