活化T细胞核因子5对人胃癌MGC803细胞增殖及凋亡能力的影响  

Effects of nuclear factor 5 of activated T cells on proliferation and apoptosis of human gastric cancer MGC803 cells

作　　者：郭隽馥[1] 于爽 王悦[1] 徐畅[1] 陆娜 周圆 GUO Junfu;YU Shuang;WANG Yue;XU Chang;LU Na;ZHOU Yuan(Teaching and Experimental Center,Liaoning University of Tra-ditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]辽宁中医药大学教学实验中心,辽宁沈阳110847

出　　处：《中国肿瘤生物治疗杂志》2018年第11期1119-1124,共6页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81803855);辽宁省教委高等学校科研基金资助项目(No.L201614)~~

摘　　要：目的:探讨活化T细胞核因子5(nuclear factor 5 of activated T cells, NFAT5)对人胃癌MGC803细胞增殖及凋亡能力的影响及其可能的机制。方法:设计并合成3条靶向NFAT5基因的siRNA(siRNA2567、siRNA2714和siRNA4562)及1条与NFAT5基因无同源性的阴性对照siRNA(NC-siRNA),脂质体介导转染人胃癌MGC803细胞后,采用Real-time PCR检测分析细胞中NFAT5 mRNA表达水平的变化,进而筛选出有效抑制NFAT5基因表达的siRNA(NFAT5-siRNA)。NFAT5-siRNA转染MGC803细胞48 h后,进一步采用Real-time PCR和Western blotting验证并检测细胞中NFAT5和S100A4 mRNA及蛋白表达水平的变化,流式细胞术和CCK-8法分析抑制NFAT5表达对细胞增殖及凋亡的影响。结果:转染siRNA2567对NFAT5 mRNA的表达抑制最为明显(P<0.01),siRNA2567被验证为NFAT5-siRNA。转染NFAT5-siRNA 48 h后,NFAT5和S100A4 m RNA及蛋白的表达水平均明显降低(P<0.05);与NC-siRNA组相比较,NFAT5-siRNA组MGC803细胞的增殖率在72 h和96 h均显著降低(P<0.01);NFAT5基因沉默48 h后,MGC803细胞的凋亡率由(2.7±0.2)%上升至(7.9±0.2)%(P<0.01)。结论:NFAT5-siRNA能有效沉默人胃癌MGC803细胞中NFAT5基因表达,在抑制细胞增殖率的同时能够有效促进细胞凋亡,该作用可能通过调控S100A4表达实现。Objective:To investigate the effects of nuclear factor 5 of activated T cells (NFAT5)on proliferation and apoptosis of hu- man gastric cancer MGC803 cells and to explore the possible mechanisms.Methods:Three siRNAs targeting NFAT5gene (siR- NA2567,siRNA2714 and siRNA4562)and one negative control siRNA were designed and chemically synthesized before transfeeted into human gastric cancer cell line MGC803 by liposome.Real-time PCR was used to detect the changes of NFAT5 mRNA expression in MGC803 cells to further pick out the siRNA that most effectively inhibit the expression of NFATS.Further,Real-time PCR and Western blotting assay were carried out to test mRNA and protein levels of NFAT5 and S100A4 in cells 48h after NFATS-siRNA transfection.Then,CCK-8assay and FCM assay were used to detect the influence of silencing NFAT5on cell proliferation and apoptosis,respectively.Results:siRNA2567 was the most effective siRNA that significantly inhibited the expression of NFAT5 mRNA (P<0.01), and thus was validated as NFAT5-siRNA.Real-time PCR and Western blotting assay confirmed that both mRNA and protein levels of NFAT5 and S100A4 were down-regulated in cells 48h after NFAT5-siRNA transfection.Compared with NC-siRNA group,the proliferation ability of MGC803 cells in the NFAT5-siRNA group was significantly down-regnlated at 72h and 96h (P<0.01).And FCM assay showed that compared with NC-siRNA group,cell apoptosis rate of NFATS-siRNA group was significantly increased from (2.7±0.2)% to (7.9±0.2)%,(P<0.01)48h after NFATS-siRNA transfection.Conclusion:NFAT5-siRNA transfection can silenee NFAT5gene expression in gastric cancer MGC803 cells effectively.NFATSmay inhibit proliferation and promote cell apoptosis of gastric cancer cells possibly through regulating S100A4 expression.

关 键 词：胃癌 MGC803细胞 NFAT5基因 S100A4基因 增殖 凋亡 

分 类 号：R730.51[医药卫生—肿瘤] R735.2[医药卫生—临床医学]