学术论文

      钙敏感受体通过PI3K/AKT通路对缺氧诱导的A549细胞增殖的影响

      Calcium-sensing receptor mediates hypoxia-induced proliferation of A549 cells through PI3K/AKT pathway

      摘要:
      目的:探讨钙敏感受体(calcium-sensing receptor,CaSR)在缺氧诱导的A549及A549/DDP细胞增殖中的信号转导途径.方法:通过缺氧培养箱内(93%N2、2%O2、5%CO2)培养24 h的方法复制细胞缺氧模型.应用Western blot技术分析PCNA及p-AKT蛋白不同处理情况下的表达水平;采用流式细胞技术检测不同处理因素对细胞周期及增殖指数的影响,应用BrdU掺入法分析不同处理因素对细胞DNA合成的影响.结果:缺氧引起A549及A549/DDP细胞PCNA及p-AKT蛋白水平上调,增加BrdU掺入量、S期细胞数量和细胞增殖指数,GdCl3能够增强缺氧的作用,但上述效应可以被LY294002抑制.结论:PI3K/AKT信号通路在缺氧活化CaSR并促进A549及A549/DDP细胞增殖中起重要作用.
      Abstract:
      AIM:To explore the cell signal transduction pathway of calcium-sensing receptor ( CaSR) media-ting hypoxia-induced proliferation of A549 cells and A549/DDP cells.METHODS: The cell proliferation was tested by BrdU incorporation assay .The cell cycle analysis was carried out by flow cytometry .The protein levels of PCNA and p-AKT were analyzed by Western blot .RESULTS: Hypoxia significantly increased the protein levels of PCNA and p-AKT, the BrdU incorporation and the cell proliferation index .GdCl3 ( an agonist of CaSR ) amplified the effect of hypoxia .LY294002 (a PI3K inhibitor) decreased the up-regulation of PCNA expression, the BrdU incorporation and the increased cell prolifer-ation index induced by hypoxia and GdCl 3 in the A549 cells and A549/DDP cells.CONCLUSION:CaSR is expressed in A549 cells and 549/DDP cells, and CaSR activation induced by hypoxia promotes the proliferation of A 549 cells and A549/DDP cells through PI3K/AKT signaling pathway .
      作者: 李光伟 [1] 苗宏志 [2] 李波 [1] 沈云虹 [3] 邱丽萍 [3] 张亚珍 [3] 金香兰 [3] 朱坤杰 [3]
      作者单位: 齐齐哈尔医学院病理生理教研室,黑龙江齐齐哈尔,161006 齐齐哈尔市第一医院胸心外科,黑龙江齐齐哈尔,161006 齐齐哈尔医学院机能实验室,黑龙江齐齐哈尔,161006
      刊 名: 中国病理生理杂志 ISTICPKU
      年,卷(期): 2017, 33(3)
      分类号: R363 R730.23
      在线出版日期: 2017年4月10日
      基金项目: 齐齐哈尔医学院博士基金资助项目