【摘要】 目的　以腺病毒做载体研究血小板第 4因子 (PF4)氨基末端改构体cDNA(p1 7 70cDNA)的荷瘤裸鼠体内、外抗血管新生作用。方法　将p1 7 70cDNA克隆至AdEasyTM系统 ,转染 2 93细胞包装成含p1 7 70cDNA的腺病毒载体。用RT PCR方法证明有p1 7 70cDNA外源基因插入 ,Western印迹分析KB细胞 (人上皮细胞癌 )表达的目的蛋白P1 7 70肽。用此病毒直接转导人脐静脉内皮细胞(HUVEC)并将此病毒注射到负有KB细胞的荷瘤裸鼠体内。以内皮细胞的增殖情况及瘤块体积、质量、瘤体内微血管数分析P1 7 70肽的抑制血管新生的作用。结果　转导p1 7 70cDNA病毒后的HUVEC增殖较含空载体病毒对照减低 58% ,注射病毒 2周后的实验组、空载体组及PBS对照组瘤块质量分别为 (0 0 86± 0 .0 54)g ,(0 .1 71± 0 .0 76)g和 (0 .1 95± 0 .0 67)g ,体积分别为 (1 6 .7± 5 .2 )mm3 、(36 .5± 2 3 .7)mm3 和 (41 .5± 1 2 .2 )mm3 ,免疫组化示微血管计数分别为 9.5± 1 .2 ,30 .6± 2 .6 ,31 .3± 2 .5 ,实验组、空载体病毒组及PBS对照组之间差异有显著性 (P <0 0 5) ,而空载体病毒与PBS两个对照组之间差异无显著性 (P >0 .0 5)。结论　腺病毒载体介导的P1 7 70肽体外具有抑制内皮细胞增殖活性 ,裸鼠体内具有抑制血管新生从而抑制肿瘤生长?更多还原

【Abstract】 Objective To investigate the in vivo effect of modi fi ed platelet factor 4(PF4)-p 17-70 cDNA on tumor angiogenesis in nude mice. Methods The p 17-70 cDNA was cloned into the AdEasy TM system to transfect packing cell line 293 and produce viral particles e ncoding p 17-70 cDNA(Ad p 17-70 ).The integration of p 17-70 cDNA was confirmed by RT-PCR and the P17-40 peptide Western blot. The biol ogical activity of purified recombinant adenovirus was determined by umbilical v einal endothelial cell proliferation assay in vitro and in vivo tumor angiogenes is suppression of nude mice bearing human head and neck carcinoma.Result s p 17-70 significantly inhibited in vitro proliferation of end othelial cells being 58% lower than that of empty vector and reduced tumor volum e in vivo. The tumor mass was (0.086±0.054)g, (0.171±0.076)g and (0.195±0.067 )g, the tumor volume was (16.7±5.2)mm 3,(36.5±23.7) mm 3 and (41.5±12.2)m m 3 in p 17-70 cDNA transfected group, empty vector group and PBS group, re spectively. Immunohistochemi cal staining demonstrated a decreased number of blood vessels in the tumors. Conclusion P 17-70 peptide mediated by adenoviral vector co uld inhibit the endothelial proliferation in vitro and the tumor growth in vivo.更多还原