沉默FGF4基因表达对肺癌A549细胞增殖及凋亡的影响  

Effect of inhibition of FGF4 gene expression by siRNA on human lung cancer cells proliferation and apoptosis

作　　者：孙凡童 李莹[1] 朱秋明[1] Sun Fantong;Li Ying;Zhu Qiuming(The First Department of Oncology,First Affiliated Hospital of Harbin Medical University,Harbin 150001,China)

机构地区：[1]哈尔滨医科大学附属第一医院肿瘤一科,150001

出　　处：《国际呼吸杂志》2020年第16期1227-1231,共5页International Journal of Respiration

摘　　要：目的研究成纤维细胞生长因子4(FGF4)下调对人肺癌A549细胞增殖与凋亡的影响及其机制。方法采用免疫荧光技术检测A549细胞中FGF4蛋白的表达,设计并合成干扰FGF4表达的小干扰RNA(siFGF4),转染人肺癌A549细胞,将细胞分为3组:空白对照组、阴性对照组和siFGF4组,收集各组转染48h细胞,CCK-8法检测细胞的增殖能力,流式细胞术检测细胞凋亡率,Hoechst33258染色观查细胞凋亡,蛋白质印迹法检测Bcl-2和Bax的表达。结果CCK-8结果显示,siFGF4组细胞增殖能力明显受到抑制(5110%±351%),与空白对照组(9905%±311%)和siNC组(9900%±359%)比较,差异有统计学意义(F=4673,P<005)。流式细胞仪检测结果显示,siFGF4组细胞凋亡率为5999%±552%,高于空白对照组(298%±224%)及siNC组(476%±205%),差异有统计学意义(F=6754,P<001)。蛋白质印迹法结果显示,与空白对照组及siNC组相比,siFGF4组Bcl-2的蛋白表达水平明显降低(F=5153,P<001),Bax的蛋白表达水平明显升高(F=4133,P<005)。结论下调人肺癌A549细胞FGF4基因表达有望为肺癌基因靶向治疗提供新的靶位。Objective To vStudy the effects of inhibition of fibroblast growth factor 4(FGF4)gene expression on human lung cancer A549 cells proliferation and apoptosis and its mechanism.Methods FGF4 protein expression in A549 cells was detected by immunofluorescence technique,small interference RNA(siFGF4)was designed and synthesised to interference of FGF4 expression,transfection in human lung cancer A549 cells,cells were devided into three group:control group,negative contrast group and siFGF4 group.The cells transfected for 48 h were collected and CCK-8 method was used to detect cell proliferation,flow cytometry was used to detect cells apoptosis rate,Hoechst 33258 dying was used to observe cells apoptosis,Western blot method was used to detect the expression of Bcl-2 and Bax.Results CCK-8 results showed that the varibility of cells in siFGF4 group was significantly inhibited(51.10%±3.51%),which was significantly different(F=46.73,P<0.05)from that in blank control group(99.05%±3.11%)and siNC group(99.00%±3.59%).The results of flow cytometry showed that the apoptosis rate of siFGF4 group was 59.99%zb 5.52%,higher than that of blank control group(2.98%±2.24%)and siNC group(4.76%±2.05%),the difference was statistically significant(F=67.54,P<0.01);Western blot results showed that compared with the blank control group and siNC group,the expression level of Bcl-2 in siFGF4 group decreased significantly(F=51.53,P<0.01),the expression level of Bax in siFGF4 group increased significantly(F=41.33,P<0.05).Conclusions Iinhibition of FGF4 gene expression is expected to provide a new targeted gene targeting treatment for the treatment of lung cancer.

关 键 词：成纤维细胞生长因子4 基因 肺癌 A549细胞 

分 类 号：R734.2[医药卫生—肿瘤]