稳定表达Angiopoietin-1基因肝癌细胞株的建立及其意义

目的 通过将Angipoietin-1基因转染培养人肝癌细胞SMMC－7721，建立长期表达Angipoietin-1基因的肝癌细胞模型。方法 基因重组构建Angipoietin-1真核表达质粒pcDNA3/Angipoietin-1,经脂质体将pcDNA3/Angipoietin-1质粒转染培养人肝癌细胞株SMMC－7721，G418筛选阳性细胞克隆，逆转录－聚合酶链反应（RT－PCR）检测转染后25-30d细胞Angipoietin-1基因mRNA表达水平。结果 （1）限制酶切和凝胶电泳证明成功构建表达质粒pcDNA3／Angipoietin-1；（2）通过脂质体Dosper将质粒pcDNA3／Angipoietin-1转染SMMC－7721，经G418筛选后获得阳性细胞克隆；（3）RT－PCR证明经脂质体转染的人肝 癌细胞株SMMC－7721可较稳定表达Angipoietin-1基因。结论 成功建立稳定表达Angipoietin-1基因的肝癌细胞株SMMC－7721／Angipoietin-1，为通过在体途径探讨Angipoietin-1对肿瘤血管生成的调节作用奠定了基础。

Construction of hepatic carcinoma cell strain stably expressing Angiopoietin-1 gene and its implication

Objectives To construct cell strain SMMC -7721/Angiopoietin-1 in which Angiopoietin-1 gene is stably expressed.Methods Angiopoietin-1 gene was inserted into the eukary otic expressing vector pcDNA3 to construct expressing plasmid pcDNA3/Angiopoieti n-1;PcDNA3/Angiopoietin-1 was transfected into hepatocellular carcinoma (HC C) cell strain SMMC-7721 by liposome Dosper and was selected with G418 for one month. The Angiopoietin-1 mRNA expression at 25 to 30 days after gene transfect ion in transfected SMMC-7721 was detected by using RT-PCR.Results (1) Restrictive digestion and gel electrophoresis proved the success ful construction of pcDNA3/Angiopoietin-1 expression plasmid;(2)PcDNA3/Angiop oietin-1 was transfected into cultured SMMC-7721 through liposome Dosper media tion, and G418 resistant cell strains were established by G418 selection, Angiop oietin-1 gene was expressed in transfected SMMC-7721/Angiopoietin-1.Conclusion HCC cell strain SMMC-7721 with stable Angiopo ietin-1 expression was succ essfully constructed, which could be applied to explore the in vivo role of Angi opoietin-1 in tumor Angiogenesis regulation.