SiRNA技术沉默成纤维细胞生长因子受体基因促进胃癌化疗的效果及其机制  

SiRNA-mediated silencing of fibroblast growth factor receptor improves chemotherapy effect in a xenograft mouse model of gastric cancer

作　　者：杨雪梅 

机构地区：[1]贵州省开阳县人民医院药剂科,贵州省贵阳市550300

出　　处：《世界华人消化杂志》2014年第19期2728-2733,共6页World Chinese Journal of Digestology

摘　　要：目的:研究siRNA基因沉默成纤维细胞生长因子受体(fibroblast growth factor receptor,FGFR)促进胃癌化疗的效果及其作用机制.方法:流式细胞仪检测人正常细胞MRC-5和胃癌细胞MGC80-3中FGFR的表达量.设计合成FGFR特异性siRNA序列,LipofectamineTM2000转入BGC-823细胞中.将实验分为对照组、顺铂组、顺铂+siRNA组及siRNA组,通过MTT法及流式细胞仪检测BGC-823细胞的增值及凋亡,Western blot检测FGFR沉默后相关蛋白表达情况.建立胃癌裸鼠模型,转染siRNA后通过荧光定量PCR检测FGFR mRNA表达量,测量不同分组(模型组、顺铂组和顺铂+siRNA组)瘤体的大小以检测治疗的效果.结果:胃癌细胞MGC80-3中FGFR的表达量明显高于人正常细胞MRC-5(P<0.05).转染siRNA后,与顺铂组及siRNA组相比,顺铂+siRNA组显著抑制细胞增殖及促进细胞凋亡(P<0.05),并且FGFR基因沉默后,细胞表面FGFR表达量减少,细胞内Caspase3及Bax表达量提高.胃癌裸鼠的瘤体在转染siRNA后,FGFR mRNA的表达量明显降低(P<0.05),且瘤体体积减小,瘤体衰退率提高(P<0.05).结论:FGFR特异性siRNA与顺铂联合使用可促进胃癌的治疗效果,为探索胃癌治疗的新方法奠定了基础.AIM： To investigate the effect of siRNA-medi-ated gene silencing of fibroblast growth factor receptor（FGFR） on the chemotherapy effect in a xenograft mouse model of gastric cancer and to explore the possible mechanism. METHODS： The expression of FGFR in MRC-5 cells and MGC80-3 cells was detected by flow cytometry. The siRNA against FGFR was con-structed and transfected into BGC-823 cells via LipofectamineTM 2000. The reduced rate of cell proliferation was assessed by MTT assay, and apoptosis rate was detected by flow cytometry. Western blot was used to detect the protein expression of FGFR, Caspase3 and Bax after gene silencing. A xenograft nude mouse model of gastric cancer was established, and fluores-cence quantitative PCR was used to detect FGFRmRNA expression levels after transfection with siRNA. Tumor size was measured to assess the effects of treatment in different groups（model group, cisplatin group, and cisplatin ＋ siRNA group）. RESULTS： The expression of FGFR in MGC80-3 cells was higher than that in MRC-5 cells（P〈0.05）. After transfection with siRNA, compared with the cisplatin group and model group, the cispla-tin ＋ siRNA group showed significantly inhibited cell proliferation and promoted apoptosis（P〈0.05）. After FGFR gene silencing, the expression of FGFR on cell surface decreased and the ex-pression levels of intracellular Caspase3 and Bax improved（P〈0.05）. The level of FGFR mRNA expression significantly decreased in tumor-bear-ing nude mice after siRNA transfection（P〈0.05）. Besides, tumor volume decreased and tumor re-cession rate increased（P〈0.05）. CONCLUSION： SiRNA-mediated FGFR inhibi-tion promotes the therapeutic effect of cisplatin against gastric cancer.

关 键 词：SIRNA 成纤维细胞生长因子受体 胃癌 顺铂 

分 类 号：R735.2[医药卫生—肿瘤]