RNA干扰胰岛素生长因子-1受体基因对人肝癌SMMC7721增殖及凋亡蛋白的影响  

Effects of RNA interference targeting insulin growth factor-1 receptor gene on proliferation and apoptosis of human hepatocellular carcinoma SMMC7721 cells

作　　者：徐冠军 储节胜 时毓 黄龙璋 傅敬忠 XU Guanjun;CHU Jiesheng;SHI Yu;HUANG Longzhang;FU Jingzhong(Department of Oncology,Jiujiang Third People'sHospital,Jiujiang,Jiangxi,332000,China)

机构地区：[1]九江市第三人民医院肿瘤科,江西九江332000

出　　处：《当代医学》2023年第30期1-7,共7页Contemporary Medicine

摘　　要：目的观察RNA干扰胰岛素生长因子-1(IGF-1)受体基因后对人肝癌细胞株SMMC7721增殖和相关凋亡因子表达的影响。方法构建两个靶向IGF-1R基因的RNA干扰真核表达质粒(IGF-1R-siRNA-1和IGF-1R-siRNA-2)分别转染至SMMC7721细胞,并设立正常对照组、空白对照组、IGF-1R-siRNA-1转染组、IGF-1R-siRNA-2转染组。采用聚合酶链反应(RT-PCR)法和Western blot检测IGF-1R基因表达变化,检测细胞增殖、侵袭、凋亡、凋亡蛋白、骨形态发生蛋白(BMPs)及相关信号通路蛋白的表达水平。结果RT-PCR结果显示,IGF-1R-siRNA-1和IGF-1R-siRNA-2表达载体转染SMMC-7721细胞后,IGF-1R基因的mRNA水平显著下调,IGF-1R-siRNA-1转染组IGF-1R基因的表达低于正常对照组,IGF-1R-siRNA-2转染组IGF-1R基因表达下调77.5%,差异有统计学意义(P<0.05)。Western blot结果显示,IGF-1R-siRNA-1转染组和IGF-1R-siRNA-2转染组IGF-1R蛋白在不同水平显著下调,IGF-1R蛋白表达抑制率分别为73.0%和81.5%,差异有统计学意义(P<0.05)。IGF-1R-siRNA-1转染组和IGF-1R-siRNA-2转染组细胞增殖率明显低于空白对照组,差异有统计学意义(P<0.05)。ImageJ软件分析划痕面积结果显示,与正常对照组和空白对照组相比,IGF-1R-siRNA-1转染组、IGF-1R-siRNA-2转染组修复率下降,差异有统计学意义(P<0.05)。Transwell实验发现,与正常对照组和空白对照组相比,IGF-1R-siRNA-1转染组、IGF-1R-siRNA-2转染组为SMMC-7721细胞侵袭能力明显下降,侵袭细胞数量明显减少,差异有统计意义(P<0.05)。流式细胞检测显示,IGF-1R-siRNA-1转染组和IGF-1R-siRNA-2转染组晚期凋亡率明显高于空白对照组,差异有统计学意义(P<0.05)。Western blot结果显示,IGF-1R-siRNA-1组、IGF-1R-siRNA-2组Bcl-2表达均明显受抑制,Bax、Cleaved Caspase-3蛋白表达量明显高于空白对照组,差异有统计学意义(P<0.05);PI3K、AKT/p-AKT、BMP-2、BMP-7蛋白表达量明显低于空白对照组,差异有统计学意义(P<0.05)。结�Objective To observe the effect of silencing insulin growth factor-1(IGF-1)gene by RNA interference on the proliferation of human hepatocellular carcinoma cell line SMMC7721 and the expression of related apoptotic factors.Methods Two RNA interference eukaryotic expres-sion plasmids(IGF-1R-siRNA-1 and IGF-1R-siRNA-2)targeting IGF-1R gene were constructed and transfected into SMMC7721 cells,respectively,the normal control group,blank control group,IGF-1R-siRNA-1 transfection group and IGF-1R-siRNA-2 transfection group were set up.The expres-sion of IGF-1R gene was detected by polymerase chain reaction(RT-PCR)and Western blot,and then the expression levels of cell proliferation,inva-sion,apoptosis,apoptotic protein,bone morphogenetic proteins(BMPs)and related signaling pathway proteins were detected.Results The results of RT-PCR showed that the mRNA level of IGF-1R gene was significantly down-regulated after transfection of IGF-1R-siRNA-1 and IGF-1R-siR-NA2 expression vectors into SMMC-7721 cells,the expression of IGF-1R gene in IGF-1R-siRNA-1 transfection group was lower than that in nor-mal control group,and the expression of IGF-1R gene in IGF-1R-siRNA-2 transfection group was down-regulated by 77.5%,the differences were statistically significant(P<0.05).Western blot results showed that IGF-1R protein was significantly down-regulated at different levels in IGF-1R-siRNA1 transfection group and IGF-1R-siRNA-2 transfection group.The inhibition rates of IGF-1R protein expression were 73.0%and 81.5%,re-spectively,and the differences were statistically significant(P<0.05).The cell proliferation rate in IGF-1R-siRNA-1 transfection group and IGF-1R-siRNA2 transfection group was significantly lower than that in blank control group,and the differences were statistically significant(P<0.05).The analysis of scratch area by ImageJ software showed that compared with the normal control group and the blank control group,the repair rate of IGF 1R-siRNA-1 group and IGF-1R-siRNA-2 group decreased,and the differences were statistica

关 键 词：胰岛素生长因子-1 肝癌SMMC7721细胞 增殖 凋亡因子 

分 类 号：R735.7[医药卫生—肿瘤]