利用RNAi技术抑制Bcl 2基因表达增强肺腺癌A549细胞放射敏感性

目的利用RNAi技术抑制Bcl 2基因的表达，观察其对肺腺癌A549细胞凋亡和放射敏感性变化的影响。方法构建Bcl 2基因的干扰质粒pBcl 2 siRNA，稳定转染人肺腺癌A549细胞，同时设立转染无意义链组、空载体组和未转染组。采用Real time RT PCR和Western blot方法, 观察Bcl 2基因在mRNA和蛋白水平表达的变化。采用流式细胞术和克隆形成方法,检测RNA干扰对A549细胞放射敏感性的影响。结果成功构建bcl 2 siRNA质粒，转染A549细胞，获得稳定转染细胞株A549/Bcl2 Ins；Bcl 2mRNA、 Bcl 2蛋白表达明显降低；A549/Bcl2 Ins凋亡率［（10.11±0.92）%］明显高于对照组，6Gy X线照射后凋亡率［（28.81±1.10）%］明显高于对照组及未照射组；A549/Bcl2 Ins的D0和Dq值分别为1.399和1.298，明显低于转染无意义序列细胞株A549/Bcl2 Neg(1.695和2.480)、转染空载体细胞株A549/SD (1.706和2.227)和空白对照细胞A549(1.816和2.777)。结论RNAi技术可以有效抑制肺腺癌A549细胞Bcl 2基因的表达，增强A549细胞对X线的放射敏感性。

RNAi inhibits Bcl-2 gene expression and enhances radio-sensitivity of lung adenocarcinoma cell line A549

Objective To explore the influence of Bcl-2 gene expression by RNAi on the radio-sensitivity of human lung adenocarcinoma cell line A549.Methods Vectors containing SiRNA targeting Bcl-2 gene were constructed and transfected into A549.Real time RT-PCR and Western blot were used to compare the efficiency of gene silencing.Flow cytometry was used to determine the effects of pBcl-2-siRNA and radiation on apoptosis.D0 and Dq of four cell lines were calculated by Clone formation array,and radio-sensitivity of A54...