shRNA-Foxp3对人胃癌BGC-823细胞增殖和凋亡的影响  

Effects of shRNA-mediated Foxp3 gene interference on proliferation and apoptosis of human gastric cancer cell line BGC-823

作　　者：杨佳华[1] 张文杰[1] 吕文杰[1] 顾钧[1] 董平[1] 项洪刚[1] 陆建华[1] 赵杨璐[1] 李茂岚[1] 吴文广[1] 翁昊[1] 刘颖斌[1] 陈磊[1] 

机构地区：[1]上海交通大学医学院附属新华医院普外科,上海200092

出　　处：《中国实用外科杂志》2013年第S1期57-60,共4页Chinese Journal of Practical Surgery

基　　金：上海市卫生局科研基金资助项目(20114246)

摘　　要：目的构建Foxp3基因的shRNA干扰表达载体,观察Foxp3基因表达对人胃癌细胞株BGC-823细胞增殖和凋亡的影响。方法根据人Foxp3序列设计并构建Foxp3基因的shRNA真核表达载体,利用脂质体LipofactamineTM2000转染胃癌细胞株,经G418筛选出稳定转染的阳性细胞,Real-time PCR检测在mRNA水平干扰质粒对Foxp3的抑制效应,MTT法、流式细胞术观察转染后细胞生长特性的变化。结果成功将含靶向Foxp3基因的短发夹双链RNA(shRNA)的重组质粒转染Foxp3表达阳性的胃癌细胞株BGC-823,转染shRNA-Foxp3重组质粒的BGC-823细胞Foxp3mRNA表达明显下降,与阴性对照组相比,差异有统计学意义(P<0.05)。转染干扰质粒抑制细胞增殖,促进细胞凋亡,与未转染组和阴性对照组相比,胃癌细胞生长周期出现阻滞在G0/G1期,差异有统计学意义(P<0.05)。结论干扰Foxp3的表达可显著抑制胃癌细胞的增殖和促进细胞凋亡。PURPOSE:To observe the effects of Foxp3 gene expression on proliferation and apoptosis of human gastric cancer cell line BGC-823 by constructing the short hairpin RNA(shRNA) for Foxp3 gene.Methods Foxp3 gene-targeting shRNA eukaryotic expression vectors was designed and constructed according to human Foxp3 sequence. Foxp3 shRNA was transfected into gastric cancer BGC-823 cells by LipofactamineTM2000 and the successfully transfected cells were selected by G418.Foxp3 mRNA was detected by real-time PCR to explore the inhibitory effect of plasmid transfection.Proliferation of BGC-823 was examined by MTT assay and apoptosis was analyzed by flow cytometry.Results A 95% Foxp3 shRNA transfection efficiency in BGC-823 cells was achieved.Real-time PCR revealed significant mRNA reduction induced by Foxp3 shRNA versus the control(P< 0.05).Proliferation was inhibited while apoptosis increased in the transfected cells,whose cell cycle was arrested in G0/G1 phase.The Results had statistical significance compared with the control group(P< 0.05).Conclusion Interference of Foxp3 expression by shRNA can significantly inhibit proliferation and induce apoptosis in gastric cancer cell line BGC-823.

关 键 词：胃癌细胞 FOXP3 SHRNA 增殖 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]