下调GPC-3基因转录对肝癌细胞侵袭及血管新生的抑制作用  

Down-regulation of glypican-3 gene transcription inhibits hepatoma cell invasion and angiogenesis

作　　者：邰伯军[1] 姚敏[2] 王理[3] 钱琦[4] 时杨[5] 蔚丹丹[4] 陆少林[5] 姚登福[5] 

机构地区：[1]南通大学附属医院感染性疾病科,江苏省南通市226001 [2]南通大学医学院免疫学教研室,江苏省南通市226001 [3]南通大学医学院医学信息学教研室,江苏省南通市226001 [4]南通大学附属医院肿瘤科,江苏省南通市226001 [5]南通大学附属医院临床医学研究中心,江苏省南通市226001

出　　处：《世界华人消化杂志》2014年第16期2221-2228,共8页World Chinese Journal of Digestology

基　　金：国家国际科技合作专项基金资助项目;No.2013DFA32150;中国博士后科学基金资助项目;No.2013M531395;江苏省博士后基金资助项目;No.苏人社发(2012)468号~~

摘　　要：目的:观察以短发夹RNA(short hairpin RNA,shRNA)下调磷脂酰肌醇蛋白多糖-3(glypican-3,GPC-3)基因转录,对抑制肝癌MHCC-97H细胞侵袭和血管新生的作用与机制.方法:将对GPC-3特异性shRNA转染肝癌MHCC-97H细胞,以荧光定量PCR和Western blot分析GPC-3 mRNA及蛋白表达;以5-ethynyl-2'-deoxyuridine(EdU)和Transwell法等评估细胞增殖、迁移和侵袭能力.结果:shRNA1转染MHCC-97H细胞,GPC-3在mRNA水平下降75.6%(t=15.473,P<0.001),蛋白表达同步下调;干预GPC-3基因转录,可抑制肝癌细胞增殖,细胞迁移与侵袭能力减低,与抗癌药物具协同作用;Wnt/β-catenin中β-catenin表达下调67.7%;Hedgehogs通路中脑胶质瘤相关癌基因1(glioma-associated oncogene 1,Gli1)表达上调53.3%;shRNA1转染72 h细胞血管内皮生长因子(vascular endothelial growth factor,VEGF)表达下调54.2%(t=46.746,P<0.001).结论:shRNA下调GPC-3转录,经Wnt/β-catenin和Hh信号通路抑制癌细胞迁移、侵袭和血管新生,为肝癌基因治疗潜在的分子靶目标.AIM： To investigate whether down-regulation of glypican-3（GPC-3） gene transcription by short hairpin RNAs（shRNAs） inhibits hepatoma MHCC-97H cell invasion and angiogenesis. METHODS： Specific GPC-3 shRNAs were transfected into MHCC-97H cells. GPC-3 mRNA and protein expression was analyzed by fluorescence quantitative PCR and Western blot, respectively. Hepatoma cell proliferation was detected by 5-ethynyl-2′-deoxyuridine and sulforhodamine B assay, and cell migration and invasion were assessed by wound healing and transwell assays. RESULTS： After MHCC-97H cells were transfected with shRNA1, GPC-3 mRNA expression was down-regulated by 75.6%（t = 15.473, P ＆lt; 0.001）, cell proliferation was inhibited significantly, and cell migration and invasion were decreased. β-catenin expression was down-regulated by 67.7%, and glioma-associated oncogene 1（Gli1） expression was up-regulated by 53.5% in MHCC-97H cells. The expression of vascular endothelial growth factor（VEGF） was significantly decreased（54.2%, t = 46.746, P ＆lt; 0.001） in cells transfected with shRNA1 compared with control cells. CONCLUSION： shRNA-mediated GPC-3 knockdown inhibits the migration, invasion and angiogenesis of hepatoma cells possibly through Wnt/β-catenin and Hh signaling pathways.

关 键 词：肝细胞性癌 磷脂酰肌醇蛋白聚糖-3 SHRNA 血管新生 

分 类 号：R735.7[医药卫生—肿瘤]