肝细胞生长因子基因转染对淋巴瘤细胞的生物学效应  

Effect of hepatocyte growth factor gene transfection on biological features of lymphoma cells

作　　者：岑东 赵行[2] 沈蓉蓉[2] 滑世轩[2] 吕建新[2] 裴仁治 涂植光[4] 

机构地区：[1]浙江省鄞州疾病预防控制中心,浙江宁波315100 [2]温州医学院浙江省医学遗传学重点实验室,浙江温州325027 [3]浙江省鄞州人民医院血液科,浙江宁波315040 [4]重庆医科大学医学检验系教育部"临床检验诊断学"重点实验室,重庆400016

出　　处：《浙江大学学报（医学版）》2010年第4期378-385,共8页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省医药卫生科技计划(2003B172;2007A175);宁波市医药卫生科技计划(2003079);宁波市科技计划(2007C10065)

摘　　要：目的:克隆肝细胞生长因子(HGF)基因,构建重组真核表达载体,并观察HGF基因转染对Raji细胞的生物学效应。方法:从人肝组织中提取总RNA,反转录后行PCR获得HGF基因cDNA,与载体pVITRO2-mcs构建重组真核表达载体,转染Raji细胞。行潮霉素B筛选,连续传代,采用实时荧光定量PCR、ELISA和细胞免疫组化及半固体培养等方法,观察HGF基因转染后的表达与对Raji细胞生物学性状的影响。结果:成功克隆HGF基因并构建了重组真核表达载体pVITRO2-mcs-HGF。经RT-PCR证实HGF基因成功转染Raji细胞。HGF基因转染后可在Raji细胞稳定表达HGF mRNA和蛋白,且可促进其增殖和迁徙及侵袭。结论:HGF基因获成功克隆并构建了重组真核表达载体,转染Raji细胞后可稳定表达,并对其生物学性状具促进作用。Objective： To investigate the biological effect of hepatocyte growth factor（HGF） on HGF gene-transfected Raji cells.Methods： Total RNA was extracted from human hepatic tissue,HGF gene cDNA was amplified by RT-PCR,and then cloned into vector pVITRO2-mcs to construct recombinant eukaryotic expression vector pVITRO2-mcs-HGF.The recombinant vector was transfected to Raji cells,and the stably transfected cells were selected by homomycin B in serial passages,and confirmed by real-time fluorescent quantitatitive PCR,ELISA,immunocytohistochemistry.The biological features of transfected Raji cells were evaluated by semisolid culture.Results： RT-PCR results showed that Raji cells were transfected successfully with recombinant eukaryotic expression vector pVITRO2-mcs-HGF.HGF mRNA and protein were expressed successfully in Raji cells.Expression of HGF gene enhanced proliferation,metastasis and invasion of Raji cells.Conclusions： HGF gene has been cloned and recombined to construct recombinant eukaryotic expression vector pVITRO2-mcs-HGF successfully.Transfected HGF may change the biological features of Raji cells.

关 键 词：肝细胞生长因子/遗传学 淋巴瘤/遗传学 基因 转染 表达 

分 类 号：R733.1[医药卫生—肿瘤]