RNA干扰HIF-1α增强人肺腺癌SPCA-1细胞放射敏感性的研究

Silencing HIF-1α through RNA interference sensitizes SPCA-1 cells

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 radiosensitization of silencing HIF-1α by RNA interference in human lung adenocarcinoma cells （SPCA-1） Methods HIF-1α RNA interference （RNAi）vector （pSUPER-HIF-1α） was constructed by using pSUPER plasmid. The RNAi effect was detected by RT-PCR and Western blot in SPCA-1 cell line, both under normoxia and hypoxia condition. The mdiosensitization effect of silencing HIF-α was measured by colony forming assay. Results After silencing the HIF-1α by RNAi, the expression of HIF-1α mR- NA was decreased to 64 % and 76 % under normoxia and hypoxia condition, respectively; the expression of HIF-1α protein was inhibited under hypoxia condition. The Do of cells tranfected with pSUPER-HIF-1α, pSUPER and controlled cell was 2.5, 5.0 and 5.1 Gy, respectively, with a radiosensitization ratio of 2.1. Conclusion RNA-interference targeting silenced HIF-1α may increase the mdiosensitivity of SPCA-1 cell line.