内皮抑素基因腺病毒载体的构建及表达

目的：构建人内皮抑素（Human 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Lipofectamine2000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Western Blot检测hE的表达；用不同感染复数（Muhiplicity of infection，MOI）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。

Study on the construction of recombinant adenovirus vector expressing human endostatin and its expressional activity

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.