GDC-0032抑制U251胶质瘤细胞活力并诱导DNA损伤  

GDC-0032 inhibits cell viability and induces DNA damage in U251 human glioma cells

作　　者：崔海娟[1] 张亚云 焦鹏[3] 孙铮 

机构地区：[1]泰山医学院护理学院,山东泰安271016 [2]郑州大学第一附属医院,河南郑州450052 [3]泰山医学院生命科学研究中心,山东泰安271000 [4]泰山医学院第二临床医学院,山东泰安271000

出　　处：《中国病理生理杂志》2017年第10期1858-1863,共6页Chinese Journal of Pathophysiology

基　　金：山东省自然科学基金资助项目(No.ZR2014HL107)

摘　　要：目的:探讨磷脂酰肌醇3-激酶(PI3K)抑制剂GDC-0032对人胶质瘤U251细胞活力、细胞周期和DNA损伤的影响。方法:GDC-0032处理U251细胞,MTT实验检测细胞活力;流式细胞术分析GDC-0032对细胞周期的影响;Western blot检测细胞内蛋白表达的变化;免疫荧光检测组蛋白γ-H2AX在细胞内的表达与分布。结果:GDC-0032剂量依赖性地抑制U251细胞活力;U251细胞经GDC-0032作用后,处于G1期的细胞数量明显增多,同时p27的表达水平增加,而细胞分裂周期蛋白2(Cdc2)的表达则受到抑制;在GDC-0032作用的细胞中,γ-H2AX焦点的生成和表达水平明显升高;另外,GDC-0032上调胶质瘤细胞中丝裂原活化蛋白激酶家族成员细胞外信号调节激酶(ERK)和c-Jun氨基末端激酶(JNK)的磷酸化水平,而survivin的表达则受到抑制。结论:GDC-0032能够通过抑制细胞活力和阻滞细胞于G1期而抑制U251细胞的增殖,并且诱导胶质瘤细胞发生DNA损伤;GDC-0032的抑癌活性与其调控ERK和JNK的活性及下调survivin的表达相关。AIM：To investigate the effect of phosphatidylinostiol 3-kinase （ PI3K） inhibitor GDC-0032 on cell viability, cell cycle and DNA damage in human glioma U 251 cells.METHODS：The cell viability was analyzed by MTT assay.The cell cycle distribution of U251 cells was examined by flow cytometry .The protein expression was examined by Western blot.The expression and localization of γ-H2AX were determined by laser-scanning confocal microscopy .RE-SULTS：GDC-0032 inhibited the cell viability in a dose-dependent manner .U251 cells showed G 1 phase arrest accompa-nied with upregulation of p 27 protein after exposure to GDC-0032, while the expression of cell division cycle protein 2 （Cdc2） was inhibited.GDC-0032 treatment increased the formation of γ-H2AX foci and histone H2AX phosphorylation in the U251 cells.In addition, GDC-0032 upregulated the phosphorylation levels of mitogen-activated protein kinases , inclu-ding extracellular signal-regulated kinase （ ERK） and c-Jun N-terminal kinase （ JNK） , in the glioma cells , while the ex-pression of survivin was inhibited .CONCLUSION：GDC-0032 inhibits U251 cell growth by inhibition of cell viability and cell cycle progression.GDC-0032 also induces DNA damage of U251 cells.The anticancer activity of GDC-0032 is associ-ated with increasing the activity of ERK and JNK and downregulating the expression of survivin .

关 键 词：GDC-0032 胶质瘤 细胞活力 DNA损伤 丝裂原活化蛋白激酶 

分 类 号：R739.4[医药卫生—肿瘤] R363[医药卫生—临床医学]