Ad-VEGI_(151)对人脐静脉内皮细胞增殖的影响  

Inhibitory effects of Ad-VEGI151 on venous endothelial cell proliferation

作　　者：刘银萍[1] 柳林[1] 潘欣[2] 徐琪[1] 潘东艳[1] 

机构地区：[1]第二军医大学长海医院眼科,上海200433 [2]第二军医大学微生物学教研室,上海200433

出　　处：《第二军医大学学报》2005年第9期1006-1008,共3页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(30271391)

摘　　要：目的:研究以复制缺陷型重组腺病毒为载体的血管内皮细胞生长抑制因子基因(VEGI151)对静脉内皮细胞的增殖抑制作用。方法:利用腺病毒载体pCA13构建携带VEGI151基因的质粒,经293A细胞包装、扩增,Western印迹法检测病变细胞内基因的蛋白质表达。X-gal染色测定重组腺病毒载体系统的基因转移效率,结晶紫染色法检测细胞D570/630值,观察Ad-VEGI151对ECV304细胞增殖抑制作用的效果,免疫组织化学检测ECV304细胞内VEGI151基因的蛋白质表达。结果:应用细胞内质粒DNA同源重组法,将脂质体介导质粒pCA13-VEGI151与pJM17共转染293A细胞制备重组腺病毒,所获病毒滴度高,是一种制备重组腺病毒切实可行的方法。Ad-VEGI151在病变细胞内能成功表达蛋白,具有较高的基因转移效率,对静脉内皮细胞的增殖具有强烈的抑制作用,并能在靶细胞内表达有生物学活性的蛋白质。结论:以复制缺陷型重组腺病毒为载体的VEGI151基因能在靶细胞内表达具有生物学活性的蛋白质,抑制体外静脉内皮细胞的增殖,这为进一步肿瘤及新生血管性疾病的基因治疗提供了新的方法。Objective：To study the inhibitory effects of adenovirus mediated vascular endothelial cell growth inhibitor 151 （VEGI151） on venous endothelial cell proliferation. Methods： VEGI151 gene was cloned into adenovirus vector pCA13 and the product was packaged and amplified with 293A cells. The expressed protein was identified by Western blot. X-gal staining was used to determine the transfection efficiency of the recombinant adenovirus system. The inhibitory effect of Ad-VEGI151 on ECV304 cell proliferation was observed and assessed by violet crystal assay. The protein expression of VEGI151 gene in ECV304 cells was detected by immunohistochemistry. Results： Liposome-mediated pCA13-VEGI151 and pJM17 cotransfection of 293A cells （through homologous recombination of intracellular plasmid） is a practical and feasible method to obtain high-titre recombinant adenovirus. The constructed Ad-VEGI151, with high transfection efficiency, significantly inhibited ECV304 cells proliferation and expressed functional protein in target cells. Conclusion： Ad-VEGI151 can express bioactive protein in target cells and can inhibit the proliferation of venous endothelial cells, and this may pave a new way for gene therapy of tumors and neovascular diseases.

关 键 词：血管内皮细胞生长抑制因子 基因治疗 腺病毒载体 

分 类 号：Q789[生物学—分子生物学]