假型逆转录病毒载体介导的tPA转染对平滑肌细胞增殖的影响  

Gene transfer of tissue plasminogen activator mediated by pseudotyped retroviral vector and its effect on smooth muscle cell proliferation

作　　者：李俊彦[1] 裴斐[1] 何蕊[1] 余红[2] 张莉[1] 

机构地区：[1]西安交通大学第二附属医院心胸外科,陕西西安710004 [2]美国迈阿密大学血管外科

出　　处：《第四军医大学学报》2008年第22期2019-2022,共4页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(30470456)

摘　　要：目的:探讨假型逆转录病毒载体VSV-G/MuLV介导组织纤溶酶原激活物(tPA),lacZ基因转染兔平滑肌细胞(SMCs)后基因的表达和对SMCs增殖的效应.方法:利用携带tPA,lacZ基因的假型逆转录病毒载体VSV-G/MuLV分别转染兔SMCs(SMCs/tPA,SMCs/lacZ).通过检测tPA抗原和测定tPA的活性观察tPA基因的表达.通过x-gal染色检测β-gal的活性测定lacZ基因的表达.用放射性标记的脱氧胸腺嘧啶苷(3H-TdR)掺入法和细胞计数法测定SMCs的增殖.结果:在没有纤溶酶原时,SMCs/tPA与SMCs在增殖上无显著差异.在有纤溶酶原时,SMCs/tPA条件培养基诱导了SMCs增殖.结论:VSV-G/MuLV介导tPA,lacZ转染到SMCs并成功表达.tPA基因转染在纤溶酶原存在的条件下促进了SMCs的增殖.AIM： To investigate the expression of the transfected tissue-type plasminoge activator （tPA）, lacZ gene mediated by pseudotyped retroviral vector VSV-G/MuLV in rabbit smooth muscle cells and test its effect on the cell proliferation. METHODS： Rabbit SMCs were transduced with pseudotyped retroviral vectors VSV-G/MuLV carrying genes coding for tPA or lacZ gene. The expression of tPA was examined by tPA antigen and activity. The activity of β-gal was determined by x-gal staining method. The SMCs proliferation was measured by a radioactive labeled thymidine incorporation assay and daily count of cell number. RESULTS： In the absence of plasminogen, no statistical difference in proliferation was observed between SMCs and SMCs/tPA. In the presence of plasminogen, SMCs/tPA enhanced significantly the SMCs proliferation. CONCLUSION： tPA, lacZ gene is successfully transduced into SMCs using VSV-C/MuLV, and is expressed successfully, tPA gene transduction significantly enhances the SMCs proliferation.

关 键 词：假型逆转录病毒载体 转染 TPA LACZ 肌 平滑/细胞学 细胞增殖 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]