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目的探讨RNA干扰乏氧诱导因子1(HIF-1)增强人肺腺癌SPCA-1细胞的放射敏感性。方法构建干扰HIF-1α质粒pSUPER-HIF-1α,采用荧光定量RT-PCR、Western blot方法观察有氧与乏氧状态下RNA干扰HIF-1α的变化,用成克隆实验观察乏氧时RNA干扰HIF-1α对放射的增敏作用。结果在常氧和乏氧的状态下,RNA干扰HIF-1α引起mRNA的表达水平分别下降64%和76%;在乏氧的状态下,蛋白的表达下降。转染pSUPER-HIF-1α后的细胞存活曲线的D0值为2.5 Gy,转染空白质粒细胞和空白对照细胞的D0值分别为5.0 Gy和5.1 Gy,增敏比为2.1。结论RNA干扰HIF-1α可以增加SPCA-1细胞对放射的敏感性。
Objective To investigate the radiosensitivity of RNA interference hypoxia inducible factor 1 (HIF-1) in human lung adenocarcinoma SPCA-1 cells. Methods The interference of HIF-1α plasmid pSUPER-HIF-1α was constructed. The changes of HIF-1α mRNA by HIF-1α were detected by real-time RT-PCR and Western blot. -1α on the radiation sensitization. Results The mRNA expression of HIF-1α decreased by 64% and 76% respectively under normoxia and hypoxia. The protein expression decreased under hypoxia. The D0 value of cell survival curve after transfection with pSUPER-HIF-1α was 2.5 Gy. The D0 values of transfected blank control cells and blank control cells were 5.0 Gy and 5.1 Gy, respectively. The sensitization ratio was 2.1. Conclusion RNA interference of HIF-1α can increase the sensitivity of SPCA-1 cells to radiation.