【摘要】 目的:构建人内皮抑素(Hum an Endostatin,hE)基因的腺病毒载体,并研究其对胃癌细胞株SGC-7901、MKN-45及人脐静脉内皮细胞系ECV304生物学特性的影响。方法:采用L ipofectam ine2000法将含有人内皮抑素基因的质粒pCA13-hE与pBGHE3共同转染293细胞;免疫荧光法及W estern B lot检测hE的表达;用不同感染复数(Mu ltip lic ity of infection,MO I)的重组人内皮抑素基因的腺病毒感染ECV304细胞,观察细胞生长。结果:成功构建了含hE基因的腺病毒载体;经含hE基因腺病毒载体感染的人SGC-7901胃癌细胞株、MKN-45胃癌细胞株均表达hE蛋白;表达的hE蛋白具有一定的生物学活性,可以抑制人静脉内皮细胞系ECV304的生长。结论:获得了有表达功能活性的Endostatin腺病毒载体,表达产物可抑制ECV304细胞的增殖,为肿瘤的抗血管基因治疗提供了必要条件。更多还原

【Abstract】 Objective:To construct the recombinant adenovirus vector expressing human endostatin and explore its biological activity which treated with the cellines of SGC-7901,MKN-45 and ECV304 by the adenovirus vector.Methods: Recombinant plasmid pcA13-hE containing human endostatin gene was cotransfered with pBGHE3 into 293 packaging cells by lipofectine-mediated transfection,the expressions of human endostatin were detected by immunofluorescence and Western-Blot,the growth behavior of ECV304 cells were obsevered when the cells were infected by different MOI recombinant adenovirus vector containing human edostatin gene. Results: The expression vector of human endostatin was successfully constructed and tranfected into the cellines of human stomach cancer SGC-7901 and MKN-45.The two cellines also expressed the human endostatin protein,and the expressional protein had biological activity and could inhibit the growth of ECV304 cells. Conclusion: The recombinant adenovirus vector of functional human endostatin gene was obtained,the expressional products inhibited the growth of ECV304 cells.The study has significance for gene therapy of tumor.更多还原