hnRNPB_1基因的RNA干扰抑制肺癌细胞A549增殖研究  

The Inhibitory Effects of siRNA Expression Vector on Cell Proliferation and Expression of hnRNPB_1 Gene in Lung Cancer A549 Cells

作　　者：蒲丹[1] 李为民[1,2] 周陶友[1] 吴迪[1] 陈小兵[1] 陈敏[1] 唐凤鸣[1] 韩娟[1] 

机构地区：[1]四川大学华西医院呼吸内科,成都610041 [2]四川大学华西医院生物治疗国家重点实验室,成都610041

出　　处：《四川大学学报（医学版）》2009年第3期389-392,共4页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号30270587);教育部博士点专项基金(20060610056)资助

摘　　要：目的探讨特异性小分子干扰（small interference，siRNA）抑制肺癌A549细胞核内不均一核糖蛋白B1（heterogeneous nuclear ribonucleoprotein，hnRNPB1）基因的表达后，对A549细胞增殖的影响。方法构建hnRNPB1特异性的siRNA真核细胞表达载体并转染人肺癌细胞株A549，观察重组载体分别在第1、4及6周对A549细胞hnRNPB1基因的干扰效果以及对肺癌细胞周期及凋亡的影响。结果特异性siRNA真核表达可显著抑制肺癌细胞hnRNPB1基因的表达，使hnRNPB1 mRNA的表达减少46％～73％，蛋白表达减少77．69％～83．04％，作用时间至少可维持克隆形成后4周，同时，hnRNPB1基因表达下调抑制了A549细胞在体外的增殖，促进了肺癌细胞的凋亡。结论特异性siRNA能特异、高效地抑制肺癌细胞株A549细胞hnRNPB1基因的表达，RNAi可能为肺癌的基因治疗提供新策略。Objective To investigate the inhibitory effects of specific siRNA on the expression of heterogeneous nuclear ribonucleoproteins （hnRNPB1） and cell proliferation in lung cancer A549 cells. Methods After the construction of siRNA expression vector for hnRNPB1, cultured A549 cells were tansfected with the specific siRNA vector, so RNA of A549 cells was interfered by RNA interference technique. The cells were retrieved at 1st, 4th and 6th week after transfeetion, cell cycles and cell apoptosis were measured with flow cytometry, the mRNAs levels of hnRNPB1 , were detected by fluorescence quantity RT-PCR, the protein levels of hnRNPB1 was detected by Western blot technique. Results It was found that plasmid-derived siRNAs could inhibit hnRNPB1 mRNA and protein expression. The expressions of hnRNPB1 mRNA and protein were decreased about 46%-73%,and 77. 69%-83. 04% respectively. We also observed that siRNA in A549 cell suppressed the cell proliferation by increasing the amounts of G1 cells, and induced cell apoptosis. Conclusion These results demonstrated the effectiveness of siRNA in lung caner cell, which indicated a possible clinical therapy method for lung cancer.

关 键 词：肺癌 核内不均一核糖核蛋白B1 小分子干扰RNA 

分 类 号：R734.2[医药卫生—肿瘤]