上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (12): 1687-.doi: 10.3969/j.issn.1674-8115.2011.12.006

• 论著（基础研究） • 上一篇    下一篇

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

马 贵, 祝 宇, 吴瑜璇, 沈周俊, 盛佳燕, 徐云泽   

  1. 上海交通大学 医学院附属瑞金医院泌尿外科, 上海 200025
  • 出版日期:2011-12-28 发布日期:2012-01-04
  • 通讯作者: 祝 宇, 电子信箱: zyyyhyq@126.com。
  • 作者简介:马 贵（1985—）, 男, 硕士生；电子信箱: timothy8581@163.com。
  • 基金资助:

    上海市教委基金(11YZ58)；上海市科委基金(09ZR1418500)

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

MA Gui, ZHU Yu, WU Yu-xuan, SHEN Zhou-jun, SHENG Jia-yan, XU Yun-ze   

  1. Department of Urology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Online:2011-12-28 Published:2012-01-04
  • Supported by:

    Shanghai Education Committee Foundation, 11YZ58；Shanghai Science and Technology Committee Foundation, 09ZR1418500

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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、2.0 μmol/L）17-AAG培养液；实验二组分别加入150 μg/L VEGF（VEGF组）、0.1 μmol/L 17-AAG（17-AAG组）、0.1 μmol/L17-AAG+150 μg/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0.1 μmol/L。0.1 μmol/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）。0.1 μmol/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。

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

Abstract:

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. Methods PC12 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. Results 17-AAG significantly inhibited the growth of PC12 cells in time- and dose-dependent manners (P<0.05), 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 (P<0.01). 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 (P<0.05). Conclusion 17-AAG, HSP90 inhibitor, can inhibit the proliferation of PC12 cells, induce cell apoptosis and inhibit the expression of VEGF protein.

Key words: PC12 cells, pheochromocytoma, heat shock protein 90, 17-allylamino-17-demethoxygeldanamycin, vascular endothelial growth factor, apoptosis