基础医学与临床 ›› 2010, Vol. 30 ›› Issue (7): 698-702.

• 研究论文 • 上一篇    下一篇

雷帕霉素抑制人多囊肾囊肿衬里上皮细胞增殖及VEGF表达

刘春艳 梅长林 袁莉 张懿 付莉莉 蔡厚安 王雪琦   

  1. 大连医科大学附属二院 第二军医大学上海长征医院 第二军医大学上海长征医院 第二军医大学上海长征医院 第二军医大学上海长征医院 第二军医大学上海长征医院
  • 收稿日期:2009-11-27 修回日期:2010-02-24 出版日期:2010-07-05 发布日期:2010-07-05
  • 通讯作者: 梅长林

Antiproliferative and antiangiogenic effect of rapamycin on ADPKD cystic lining epithelial cells

Chun-yan LIU, Chang-lin MEI, Li YUAN, Yi ZHANG, Li-li FU, Hou-an CAI, Xue-qi WANG   

  1. the second affiliated hospital of Dalian medical university Changzheng Hospital, Second Military Medical University
  • Received:2009-11-27 Revised:2010-02-24 Online:2010-07-05 Published:2010-07-05
  • Contact: Chang-lin MEI,

摘要: 目的 探讨雷帕霉素对常染色体显性多囊肾病囊肿衬里上皮细胞增殖及血管内皮细胞生长因子(VEGF)表达的抑制作用及其机制。方法MTT法检测WT9-12细胞增殖;流式细胞术检测细胞周期及凋亡;Western Blot检测周期相关蛋白(cyclinD、p21)、凋亡相关蛋白(Bcl2/Bax)及VEGF表达。结果 雷帕霉素可抑制WT9-12细胞的增殖,使细胞周期停滞在G0/G1期并促进细胞凋亡。雷帕霉素可下调WT9-12细胞cyclinD 、上调p21表达,下调Bcl2、上调Bax表达。原代培养的多囊肾囊肿衬里上皮细胞及WT9-12细胞的VEGFmRNA表达明显高于正常肾小管上皮细胞(P<0.05)。雷帕霉素可抑制WT9-12细胞VEGF的表达(P<0.05)。结论 雷帕霉素可通过抑制多囊肾囊肿衬里上皮细胞增殖、促进凋亡及抑制VEGF表达抑制血管形成,从而抑制多囊肾病进展。

关键词: 常染色体显性多囊肾病, 雷帕霉素, 细胞周期, 凋亡, VEGF

Abstract: Objective To investigate antiproliferative effect and reduction of VEGF expression of rapamycin on ADPKD cystic lining epithelial cells. Methods ADPKD cystic lining immortalized epithelial cells(WT9-12) were stimulated by rapamycin with different concentrations. After treatment, MTT method was performed to detect the levels of proliferation, the mRNA expressions of VEGF were measured by Real-time PCR. Western blot method was used to detect the protein expressions of cyclinD, P21, Bcl2, Bax and VEGF. Results Rapamycin , a mTOR inhibitor, was found to induce cell growth inhibition, apoptosis as well as a stable accumulation of cells in the G0/G1 phase. Western blot of cultured WT9-12 cells demonstrated that Rapamycin downregulated cyclinD, Bcl2 and upregulated p21, Bax expressions. In addition, realtime-PCR detected elevated levels of VEGF mRNA in primary cells from ADPKD patients and WT9-12 cells compared with normal tubule cells. Rapamycin downregulated VEGF expression in WT9-12 cells. Conclusion Rapamycin induces ADPKD cystic lining epithelial cells growth inhibition, apoptosis and inhibits VEGF expression, which may be one of mechanisms of rapamycin postponing ADPKD progression.

Key words: ADPKD, Rapamycin, Cell cycle, Apoptosis, VEGF

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