Abstract
背景与目的肺癌耐药是肺癌患者死亡的主要原因，PPAR-γ可促进细胞凋亡，逆转肺癌耐药。本实验旨在探讨PPAR-γ表达下调对人肺癌A549顺铂耐受性和细胞凋亡的影响。方法构建siRNA沉默PPAR-γ的A549细胞系[A549/PPAR-γ(-)]，应用MTT法检测顺铂对PPAR-γ沉默A549细胞增殖的影响，应用流式细胞术检测顺铂对PPAR-γ沉默A549细胞周期的影响，Western blot法检测磷酸化Akt（p-Akt）、caspase-3和bcl-2/bax的变化，最后以RT-PCR检测bcl-2的转录水平。结果成功构建出两个A549/PPAR-γ(-)细胞克隆，经RT-PCR和Western blot检测其PPAR-γ的水平明显下降。PPAR-γ沉默后，两个克隆A549细胞对顺铂的耐受性分别增加了1.29倍和1.60倍，肿瘤细胞的凋亡减少。Western blot检测显示Akt的磷酸化水平和bcl-2/bax水平升高，caspase-3表达降低，RT-PCR进一步显示bcl-2的转录水平升高。结论抑制A549中PPAR-γ的表达后，肿瘤细胞获得对顺铂药物更高的耐受性，其机制与升高Akt磷酸化水平和bcl-2表达水平，抑制细胞凋亡有关。PPAR-γ下调是临床肿瘤产生耐药性的可能机制之一。
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