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,高家让,田卫东,李声伟,刘磊,付春华
Author：GAO Zhen- nan, GAO Jia-rang, TIAN Wei-dong, LI Sheng-wei, LIULei, FU Chun-hua
收稿日期：2005-04-25 年卷（期）页码：2005,23(02):170-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：人肿瘤坏死因子-α,基因转染,人胚成肌细胞,基因表达,
Key words：human tumor necrosis factor-α,gene transfection,human embryo myoblasts,gene expression,
基金项目：四川省卫生厅科研基金资助项目(1999年)
中文摘要
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、96 h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、96 h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
英文摘要
Objective To observe human tumor necrosis factor-α(hTNF-α) expression and secreting level of human embryo myoblasts transfected by hTNF-αgene.Methods Human embryo myoblasts were transfected with shuttle plasmid pSV23SHTNF containing hTNF-αgene by cationic liposomes DOSPER. The control group was only given equivalent liposomes except plasmid. After culturing for 24, 48, 72 and 96 hours, hTNF-αexpression level of human embryo myoblastswas observed with immunocyto- chemistry staining, and hTNF-αsecreting of human embryo myoblasts was analyzed by ELISA.Results After transfected by hT- NF-αgene for 24 , 48, 72 and 96 hours, the human embryomyoblasts displayed significant secretion of hTNF-αin the cultural su- pernatant(P
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