血管内皮生长因子激酶功能区受体基因沉默抑制肺癌A549细胞增殖并增强其对多西他赛的敏感性

Kinase-domain insert containing receptor gene silencing inhibited proliferation of human lung cancer cell line A549 and enhanced their sensitivity to docetaxel

目的研究沉默血管内皮生长因子激酶功能区受体(kinase.domain insert containing receptor,KDR)基因对人肺癌A549细胞增殖以及对化疗药物多西他赛敏感性的影响。方法设计合成:KDR的小干扰RNA(small interfering RNA,siRNA)序列,Lipofectamine^(TM)2000转染入A549细胞。通过反转录-聚合酶链反应和Western Blot检测KDR基因沉默后KDR mRNA及蛋白的表达情况,利用流式细胞仪检测A549细胞的周期变化。采用四甲基偶氮唑盐比色法及细胞克隆形成实验观察,沉默KDR基因后A549细胞对多西他赛的敏感性。结果KDR基因经48 h沉默后,A549细胞的KDR基因和蛋白的表达出现较明显的下降(P<0.05)。A549细胞的周期在G0/G1期阻滞,S期细胞数目减低(P<0.05)。在KDR基因沉默组,A549细胞对多西他赛的敏感性有明显的增强(P<0.05)。结论KDR-siRNA能够明显沉默A549细胞KDR基因和蛋白的表达,并能抑制A549细胞的增殖,增强其对多西他赛的敏感性。

Objective This study investigated the effect of small interfering RNA-mediated kinase-domain insert containing receptor(KDR) knock-down on proliferation of human lung cancer cell line A549 and their sensitivity to docetaxel.Methods The small interfering RNA(siRNA)against KDR was constructed and transfected into A549 cells with Lipofectamine 2000.The expression of KDR was detected by reverse transcription-polymerase chain reaction and Western Blot.Flow cytometry was used to detect the cell cycle.Sensitivity to docetaxel after transfection were examined by methyl thiazolyl tetrazolium assay and clonogenic assay.Results In A549 cells,the protein and mRNA levels of KDR were decreased significantly after transfection,and reduction of proliferation was related to an increase in the fraction of GO/Gl phase.The sensitivity of A549 cells to docetaxel was increased significantly after transfection.Conclusion The KDR special siRNA silenced KDR,decreased A549 cells proliferation and enhanced their sensitivity to docetaxel.