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.
出处
《第四军医大学学报》
CAS
北大核心
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)
