人肿瘤坏死因子-α基因转染人胚成肌细胞的表达和分泌研究  

Expression and Secretion of Human Tumor Necrosis Factor Gene Transfected on Human Embryo Myoblasts

作　　者：高振南[1] 高家让[1] 田卫东[1] 李声伟[1] 刘磊[1] 付春华[1] 

机构地区：[1]四川大学口腔生物医学工程教育部重点实验室,四川成都610041

出　　处：《华西口腔医学杂志》2005年第2期170-172,共3页West China Journal of Stomatology

基　　金：四川省卫生厅科研基金资助项目(1999年)

摘　　要：目的应用人肿瘤坏死因子_α(hTNF_α)基因转染人胚成肌细胞,观察转染hTNF_α基因后人胚成肌细胞表达和分泌hTNF_α的水平。方法用阳离子脂质体DOSPER介导含hTNF_α基因的穿梭质粒pSV23SHTNF转染人胚成肌细胞;对照组只给予等量的脂质体,不加入质粒。转染基因后24、48、72、96h用免疫组织化学染色观察hTNF_α基因在人胚成肌细胞细胞浆和细胞膜的表达水平,并用ELISA法检测人胚成肌细胞培养上清液中hTNF_α的浓度。结果免疫组织化学染色发现实验组人胚成肌细胞在转染hTNF_α基因后24、48、72、96h均见细胞浆和细胞膜hT NF_α呈强阳性表达,而对照组均呈弱阳性;ELISA检测发现实验组人胚成肌细胞培养上清液中hTNF_α平均浓度在各时期均高于对照组(P<0.05),尤以转染72h后最高,达401.0pg/ml。结论hTNF_α基因转染成肌细胞,可使成肌细胞持续高表达和高浓度分泌hTNF_α,提示可以用hTNF_α基因转染舌癌周围肌细胞,使癌周微环境肌细胞持续表达和分泌高浓度hTNF_α来抑制舌癌向肌深层浸润。Objective To observe human tumor necrosis factor-α (hTNF-α) expression and secreting level of human embryo myoblasts transfected by hTNF-α gene.Methods Human embryo myoblasts were transfected with shuttle plasmid pSV23SHTNF containing hTNF-α gene by cationic liposomes DOSPER. The control group was only given equivalent liposomes except plasmid. After culturing for 24, 48, 72 and 96 hours, hTNF-α expression level of human embryo myoblasts was observed with immunocytochemistry staining, and hTNF-α secreting of human embryo myoblasts was analyzed by ELISA.Results After transfected by hTNF-α gene for 24 , 48, 72 and 96 hours, the human embryo myoblasts displayed significant secretion of hTNF-α in the cultural supernatant(P<0.05), and overexprsssion in cytoplasma and cell membrane.Conclusion Transfection of hTNF-α gene to human myoblasts made myoblasts secrete high concentration of hTNF-α, implying it is feasible that transfecting muscle cells surrounding tongue carcinoma lesion with hTNF-α gene can prevent tongue carcinoma from intruding into deeper muscle tissue.

关 键 词：人肿瘤坏死因子-Α 基因转染 人胚成肌细胞 基因表达 

分 类 号：R739.8[医药卫生—肿瘤]