17-丙烯胺-17去甲氧格尔德霉素对大鼠嗜铬细胞瘤细胞凋亡及血管内皮生长因子表达的影响  

Effects of 17-allylamino-17-demethoxygeldanamycin on apoptosis and expression of vascular endothelial growth factor in pheochromocytoma cells

作　　者：马贵[1] 祝宇[1] 吴瑜璇[1] 沈周俊[1] 盛佳燕[1] 徐云泽[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院泌尿外科,上海200025

出　　处：《上海交通大学学报（医学版）》2011年第12期1687-1691,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金：上海市教委基金(11YZ58);上海市科委基金(09ZR1418500)~~

摘　　要：目的探讨热休克蛋白90(HSP90)抑制剂17-丙烯胺-17去甲氧格尔德霉素(17-AAG)对大鼠嗜铬细胞瘤细胞系PC12细胞生长及血管内皮生长因子(VEGF)表达的影响。方法将实验组PC12细胞分成两组:实验一组分别加入不同终浓度(0.005、0.025、0.05、0.1、0.25、0.5、1.0、2.0μmol/L)17-AAG培养液;实验二组分别加入150μg/L VEGF(VEGF组)、0.1μmol/L17-AAG(17-AAG组)、0.1μmol/L17-AAG+150μg/L VEGF(17-AAG+VEGF组)培养液。另设DMSO组(阴性对照组)和空白对照组。采用MTT法检测细胞存活率;Wrights-Giemsa染色观察细胞形态变化;流式细胞术检测细胞凋亡率;Western blotting检测细胞中VEGF-165蛋白的表达。结果 17-AAG呈时间、剂量依赖性抑制PC12细胞增殖(P<0.05);24 h半数抑制浓度(IC50)为0.1μmol/L。0.1μmol/L 17-AAG作用于PC12细胞6、12、24、48 h的细胞凋亡率均明显高于空白对照组(P<0.01)。0.1μmol/L 17-AAG作用于PC12细胞6、12、24、48 h,VEGF-165蛋白表达逐渐降低;各时点VEGF-165蛋白表达量与阴性对照组比较,差异均有统计学意义(P<0.05)。结论 HSP90抑制剂17-AAG能抑制PC12细胞增殖,诱导细胞凋亡,抑制VEGF蛋白表达。Objective To investigate the effects of 17-allylamino-17-demethoxygeldanamycin（17-AAG）,heat shock protein 90（HSP90） inhibitor,on the growth and expression of vascular endothelial growth factor（VEGF） in pheochromocytoma cell line PC12 in rats. MethodsPC12 cells in experiment group were divided into experiment group 1 and experiment group 2.PC12 cells in experiment 1 were treated with different concentrations（0.005 μmol/L,0.025 μmol/L,0.05 μmol/L,0.1 μmol/L,0.25 μmol/L,0.5 μmol/L,1.0 μmol/L and 2.0 μmol/L） of 17-AAG culture fluid respectively,and those in experiment group 2 were treated with 150 μg/L VEGF culture fluid（VEGF group）,0.1 μmol/L 17-AAG culture fluid（17-AAG group） and 0.1 μmol/L17-AAG＋150 μg/L VEGF culture fluid（17-AAG＋VEGF group） respectively.Besides,DMSO group（negative control group） and blank control group were also established.Cell survival rate was measured by MTT assay,cell morphology was observed by Wrights-Giemsa staining,cell apoptosis was detected by flow cytometry,and expression of VEGF-165 protein in cells was determined by Western blotting. Results17-AAG significantly inhibited the growth of PC12 cells in time-and dose-dependent manners（P0.05）,with 50% inhibitory concentration（IC50） of 0.1 μmol/L.The apoptosis rates of PC12 cells after treatment with 0.1 μmol/L 17-AAG for 6 h,12 h,24 h and 48 h were significantly higher than that in blank control group（P0.01）.The expression of VEGF-165 protein gradually decreased with treatment of PC12 cells with 0.1 μmol/L 17-AAG for 6 h,12 h,24 h and 48 h,and the expression of VEGF-165 protein at each time point was significantly different from that in negative control group（P0.05）. Conclusion17-AAG,HSP90 inhibitor,can inhibit the proliferation of PC12 cells,induce cell apoptosis and inhibit the expression of VEGF protein.

关 键 词：PC12细胞 嗜铬细胞瘤 热休克蛋白90 17-丙烯胺-17去甲氧格尔德霉素 血管内皮生长因子 细胞凋亡 

分 类 号：R96[医药卫生—药理学]