组织型纤溶酶原激活剂基因转染AGZY83-a细胞移植入小鼠体内的表达研究

In vivo expression of human tissue-plasminogen activator transfected AGZY83-a cells implanted in mice

目的 探讨人组织型纤溶酶原激活剂 (tissue- plasminogen activator,t- PA)基因转染AGZY83- a细胞 (AGZY83- a/ t PA)在动物体内表达的稳定性 ,以及通过细胞移植方法进行 t- PA基因治疗的可行性。方法 将转染 t- PA c DNA的 AGZY83- a细胞在 G418的严格筛选下体外培养 ,收集细胞制成悬液 ,注射入小鼠体内不同部位 ,定期检测小鼠血浆中 t- PA的含量。结果 移植后 ,小鼠血浆中 t- PA的含量与移植前及对照组比较差异有显著性 (P<0 .0 1) ,并且至少可持续 10 5天。其中腹腔注射组表达水平最高 ,皮下注射组次之 ,两者均明显高于股四头肌注射组。结论 转 t- PA基因 AGZY83- a细胞移植于小鼠体内可稳定表达 t- PA,并且通过腹腔和皮下植入效果好于骨骼肌植入 ,为临床通过细胞移植进行 t-

Objective: To detect the expressing levels of human tissue-plasminogen activator(t-PA) in AGZY83-a cells transfected with pcDNA3.1 (+) t-PA in vivo and the feasibility of using transplantation of cells for gene therapy of thrombotic diseases. Methods: Expression vectors containing the t-PA cDNA gene were transfected into AGZY83-a cells. The transfected AGZY83-a cells were implanted into mice in different regions, and the plasma levels of human t-PA were assayed at intervals. Results: The plasma levels of human t-PA were significantly increased in mice after implantation of transfected AGZY83-a cells and were significantly higher than those of control groups implanted with untransfected AGZY83-a cells. This significant increase lasted at least 105 days. The intraperitoneal implantation group expressed the highest level of human t-PA, a little higher than that of the subcutis implantation group, while both are much higher than that of the quadriceps femoris implantation group which expressed the lowest. Conclusion: The implanted transfected AGZY83-a cells are able to stably express high levels of human t-PA, and transplantation of cells transfected with pcDNA3.1 (+) t-PA is a new promising method for gene therapy of thrombotic diseases.