慢病毒介导过表达果糖-1,6-二磷酸酶1基因降低胃癌细胞糖酵解水平的机制研究  

Research on the mechanism of lentivirus mediated overexpression of fructose-1,6-two phosphatase 1 gene reducing glycolytic level in gastric cancer cells

作　　者：王春芳 刘兵[1] 孙光[1] 徐瑜杰 赵国栋 WANG Chunfang;LIU Bing;SUN Guang;XU Yujie;ZHAO Guodong(Department of Gastrointestinal Surgery,Affiliated Haikou Hospital of Central South University Xiangya School of Medicine,Haikou 570208.Chin)

机构地区：[1]中南大学湘雅医学院附属海口医院胃肠外科,海南海口570208

出　　处：《胃肠病学和肝病学杂志》2018年第8期850-854,共5页Chinese Journal of Gastroenterology and Hepatology

摘　　要：目的探讨慢病毒介导过表达果糖-1,6-二磷酸酶1(FBP1)基因对胃癌SGC7901细胞糖酵解的影响及其机制。方法 RT-PCR和Western blotting检测人正常胃黏膜上皮GES-1细胞和胃癌SGC7901细胞中FBP1 mRNA和蛋白表达。以慢病毒介导的p Lenti6.3/FBP1质粒转染SGC7901细胞,采用RT-PCR和Western blotting检测转染效果,CCK-8和Transwell实验检测各组细胞的增殖和迁移能力,Western blotting检测缺氧诱导因子1α(HIF-1α)和葡萄糖转运蛋白1(GLUT-1)蛋白的表达,生物化学法检测胞外乳酸含量,酶联免疫吸附试验(ELISA)检测胞内磷酸果糖(PFK)含量。结果与GES-1细胞相比,SGC7901细胞中FBP1mRNA和蛋白表达明显升高(P<0.05)。p Lenti6.3/FBP1质粒转染使SGC7901细胞中FBP1 mRNA和蛋白表达显著升高(P<0.05),显著抑制SGC7901细胞的增殖和迁移能力(P<0.05);同时,降低了HIF-1α和GLUT-1蛋白的表达以及胞内PFK和胞外乳酸含量(P<0.05)。结论 FBP1过表达能够通过降低细胞的糖酵解水平抑制胃癌SGC7901细胞的增殖和迁移,其作用机制可能与抑制GLUT-1和HIF-1α表达有关。Objective To investigate the effect of lentivirus mediated overexpression of fructose-1,6-two phosphatase1（FBP1） gene on glucolysis degradation in gastric cancer SGC7901 cells and its mechanism. Methods The expression of FBP1 mRNA and protein in human normal gastric epithelial GES-1 cells and gastric cancer SGC7901 cells were detected by RT-PCR and Western blotting. SGC7901 cells were infected with p Lenti6. 3/FBP1 plasmid mediated by lentivirus,and the transfection effect was tested by RT-PCR and Western blotting. The proliferation and migration ability of the cells were measured by CCK-8 and Transwell. The expressions of hypoxia inducible factor 1 alpha（HIF-1α） and glucose transporter 1（GLUT-1） proteins were detected by Western blotting. The content of extracellular lactate was detected by biochemical method and the content of fructose phosphate（PFK） was detected by enzyme linked immunosorbent assay（ELISA）. Results Compared with GES-1 cells,the expressions of FBP1 mRNA and protein in SGC7901 cells were increased significantly（P〈0. 05）. The expressions of FBP1 mRNA and protein in SGC7901 cells were increased significantly after p Lenti6. 3/FBP1 plasmid infection（P〈0. 05）,and the proliferation and migration of SGC7901 cells were inhibited significantly（P〈0. 05）,and the expressions of HIF-1α,GLUT-1 proteins,and the content of intracellular PFK and extracellular lactate were decreased observably（P〈0. 05）. Conclusion FBP1 overexpression can inhibit proliferation and migration of gastric cancer SGC7901 cells by decreasing the level of glucose degradation,and the mechanism may be related to the inhibition of the expressions of GLUT-1 and HIF-1α.

关 键 词：胃癌 果糖-1 6-二磷酸酶1 糖酵解 细胞增殖 迁移 

分 类 号：R735.2[医药卫生—肿瘤]