腺病毒介导的TSP1_f基因转移对脐静脉内皮细胞株EC V304增殖的影响  

Effect of TSP1_f gene transfer on proliferation of human umbilical vein endothelial cells ECV304

作　　者：刘澎[1] 李建勇[1] 陆化[1] 韩忠朝[2] 

机构地区：[1]南京医科大学第一附属医院血液科,210029 [2]中国医学科学院中国协和医科大学血液学研究所

出　　处：《江苏医药》2005年第10期735-737,共3页Jiangsu Medical Journal

基　　金：国家高技术研究发展计划(863计划;2003AA205060);南京医科大学创新基金(CX2003004)

摘　　要：目的探讨腺病毒介导的凝血酶敏感蛋白1(TSP1)抗血管新生衍生物(TSP1f)基因转移对人脐静脉内皮细胞株ECV304增殖的影响。方法应用RT-PCR方法从正常人外周血单个核细胞扩增TSP1fcDNA序列构建TSP1f重组腺病毒(ADV-TSP1f);Western blot方法检测ADV-TSP1f介导TSP1f在K562细胞中的表达;台盼蓝拒染检测经ADV-TSP1f感染的K562细胞条件培养基对ECV304细胞增殖的抑制作用。结果K562细胞经ADV-TSP1f感染可高效表达/分泌TSP1f多肽;经ADV-TSP1f感染的K562细胞条件培养基可显著抑制ECV304细胞增殖,以ADV-LacZ及PBS作对照,抑制率分别为51.6%和53.8%。结论ADV-TSP1f可显著抑制人脐静脉内皮细胞株ECV304增殖,可望成为有效的恶性肿瘤血管新生抑制剂。Objective To explore the effect of TSP1f gene transfer on proliferation of human umbilical vein endothelial cells ECV304. Methods TSP1f cDNA was amplified by RT-PCR from norreal human peripheral blood mononuclear cells and used as transgene to construct an adenoviral vector （ADV-TSP1f）. TSP1f expression/secretion by ADV-TSP1f infected K562 cells was demonstrated using Western blot analysis. The functional validation of adenovirus-mediated TSP1f was determined by an in vitro human umbilical vein endothelial cell ECV304 proliferation inhibition assay. Results TSP1f was expressed and secreted efficiently by ADV-TSPlf-infected K562 cells. Conditioned media from ADV-TSP1f infected cells inhibited ECV304 proliferation by 51.6% （compared with conditioned media from ADV-LacZ-infected cells）and 53.8% （compared with conditioned media from uninfected cells ）. Conclusion Adenovirus-mediated TSP1f gene transfer can greatly inhibit ECV304 proliferation and may serve as an effective tumor angiogenesis inhibitor.

关 键 词：凝血酶敏感蛋白1 血管新生 基因治疗 腺病毒 ECV304细胞增殖 内皮细胞株ECV304 腺病毒介导 人脐静脉 基因转移 K562细胞 

分 类 号：R331.36[医药卫生—人体生理学] R322.123[医药卫生—基础医学]