解剖学报

解剖学报 ›› 2018, Vol. 49 ›› Issue (2): 198-203.doi: 10.16098/j.issn.0529-1356.2018.02.010

• 肿瘤生物学 • 上一篇    下一篇

17-烯丙胺基-17-去甲氧基格尔德霉素对人绒毛膜癌JAR细胞系细胞周期和凋亡的影响及其相关机制

谷苗 巴一 崔丽军 刘百艺 安国庆 李玉红 许倩*   

  1. 承德医学院基础医学院，承德医学院生物医学工程系，河北 承德 067000
  • 收稿日期:2017-07-03 修回日期:2017-09-13 出版日期:2017-04-06 发布日期:2018-04-06
  • 通讯作者: 许倩 E-mail:xuqian@163.com
  • 基金资助:
    河北省高校重点学科建设项目;河北省计生委项目;河北省教育厅项目

Effect of 17-allylamino-17-demethoxygeldanamycin on cell cycle and apoptosis of JAR cells and the associated mechanisms

GU Miao BA Yi CUI Li-jun LIU Bai-yi AN Guo-qing LI Yu-hong XU Qian*   

  1. Department of Basic Medicine, Department of Biomedical Engineering, Chengde Medical College，Hebei Chengde 067000，China
  • Received:2017-07-03 Revised:2017-09-13 Online:2017-04-06 Published:2018-04-06
  • Contact: XU Qian E-mail:xuqian@163.com

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摘要：

目的 探讨热休克蛋白 90（HSP90）抑制剂17-烯丙胺基-17-去甲氧基格尔德霉素（17-AAG）对人绒毛膜癌JAR细胞系细胞周期和凋亡的影响及其相关机制。方法 体外培养人绒毛膜癌JAR细胞系，经不同浓度的17-AAG作用24 h后，采用TUNEL细胞凋亡法及Annexin V-FITC/PI双标记流式细胞术检测细胞凋亡；流式细胞术检测细胞周期；Real-time PCR法和Western blotting法检测血管内皮生长因子（VEGF）、cyclinD1、cyclinA1和 Caspase-3在mRNA和蛋白水平的表达情况。 结果 17-AAG对JAR细胞生长具有明显的抑制作用，并存在浓度依赖性。各组细胞发生G2期阻滞及明显凋亡；5、10、20 mg/L 17-AAG 作用24 h 后，各组细胞增殖相关基因cyclinD1 mRNA及蛋白表达水平相对于0 mg/L 17-AAG处理组（对照组）均下调，VEGF蛋白表达水平相对于0 mg/L 17-AAG处理组（对照组）下调，且下调程度与处理浓度成正比，而凋亡相关基因Caspase-3的mRNA未出现明显上调，但其在蛋白表达水平上调。 结论 17-AAG能够抑制JAR细胞增殖活力，诱导细胞凋亡，17-AAG 可能通过下调VEGF 的表达，产生抗肿瘤血管新生作用；可能通过下调cyclinD1使细胞周期发生G2期阻滞；且可能通过上调Caspase-3诱导细胞凋亡。

关键词: 17-烯丙胺基-17-去甲氧基格尔德霉素, 人绒毛膜癌JAR细胞, 血管内皮生长因子, 流式细胞术, 免疫印迹法

Abstract:

Objective To explore the effects of 17-AAG on cell cycle and apoptosis of Choriocarcinoma cells JAR and to clarify the related mechanisms． Methods Choriocarcinoma cells JAR were cultured in vitro and incubated with 17-allylamino-17-demethoxygeldanamycin (17-AAG) at different concentrations for 24 hours. The apoptosis of JAR cells with different concentrations of 17-AAG was measured by TUNEL analysis and flow cytometry (Annexin V-FITC/PI). Cell cycle was analyzed by flow cytometry. Real-time PCR and Western blotting analysis were used for the detection of vascular endothelial growth factor(VEGF), cyclinD1,cyclinA1 and Caspase-3 mRNA levels and protein levels. Results 17-AAG had obvious inhibitory effect on JAR cell growth with concentration dependent. When the 17-AAG concentration increased, G2 phase retardation and obvious apoptosis were found in every groups. Real-time PCR revealed an decreased expression level of cyclinD1. Western blotting analysis revealed an decreased expression level of VEGF, cyclinD1 and an increased expression level of Caspase-3. Conclusion 17-AAG can inhibit the JAR cell proliferation activity, induce cell apoptosis, and may exert the effect by down-regulation of VEGF, cyclinD1 and up-regulation of Caspase-3.

Key words: 17-allylamino-17-demethoxygeldanamycin, Choriocarcinoma cells JAR, Vascular endothelial growth factor, Flow cytometry, Western blotting