siRNA干扰沉默bFGF基因对肺腺癌A549增殖和凋亡的影响  

Effect of bFGF-siRNA-mediated gene silencing on proliferation and apoptosis in the lung cancer cell line A549

作　　者：孟又胜[1] 王秀问[1] 王亚伟[1] 田甜甜[1] 

机构地区：[1]山东大学齐鲁医院肿瘤防治研究中心,济南250012

出　　处：《山东大学学报（医学版）》2012年第3期50-54,共5页Journal of Shandong University:Health Sciences

摘　　要：目的利用小分子RNA干扰沉默碱性成纤维细胞生长因子(bFGF)基因的表达,探讨其对肺腺癌A549细胞Oct-4表达、细胞增殖和凋亡的影响。方法将A549细胞分为转染干扰质粒组(干扰质粒组)、转染阴性对照质粒组(阴性对照组)和未转染质粒组(空白对照组)。采用Real-Time PCR检测质粒干扰后A549细胞bFGF基因mRNA表达的变化;Western Blot检测Oct-4蛋白表达的变化;CCK-8比色分析法绘制生长曲线观察细胞增殖能力变化及Annexin-V-PI双染流式细胞术检测细胞凋亡。结果 Real-Time PCR结果显示质粒干扰后A549细胞中bFGFmRNA表达下降(P<0.01);Western Blot显示干扰组A549细胞Oct-4蛋白表达下降(P<0.01);生长曲线结果显示干扰质粒组细胞增殖活力下降明显(第2～4天P<0.0 5,第6～7天P<0.01);流式细胞仪检测凋亡结果显示细胞凋亡增加(P<0.0 5)。结论靶向bFGF基因的小分子RNA可以抑制bFGF表达;bFGF基因被抑制后下调Oct-4表达,细胞凋亡增加,增殖能力下降。Objective To investigate the effect of bFGF-siRNA-mediated gene silencing on expression of Oct-4,cell proliferation and apoptosis in the lung cancer cell line A549.Methods A549 cells were divided into three groups： the PGCsiU6-bFGF-siRNA group（transfected with the bFGF interference plasmid）,the negative control group（transfected with the PGCsiU6-NC-siRNA plasmid）and the blank control group（non-transfected group）.The bFGF mRNA levels were detected by real-time PCR and Oct-4 protein levels were detected by West Blot.Cell growth ability was determined by CCK-8 colorimetry.Cell apoptosis was assayed by flow cytometry.Results Specific bFGF-siRNA down-regulated the mRNA levels of the bFGF gene（P0.01） and expression of Oct-4 decreased（P0.01）.Compared with the negative and blank control groups,cell proliferation decreased in the PGCsiU6-bFGF-siRNA group（day 2-4 P0.05,day 5-7 P0.01）,while the ratio of apoptosis increased after transfection with the bFGF interference plasmid（P0.05）.Conclusion bFGF-siRNAs can remarkably down-regulate bFGF expression at the transcriptional level,down-regulate the Oct-4 expression,inhibit cell growth and induce cell apoptosis in the lung cancer cell line A549.

关 键 词：肺肿瘤 RNA干扰 碱性成纤维细胞生长因子 Oct-4蛋白 增殖 细胞凋亡 

分 类 号：R730.54[医药卫生—肿瘤]