【摘要】 背景:转染胞嘧啶脱氨酶基因的骨髓间充质干细胞能有效地将化疗前药5-氟尿嘧啶转化成具有细胞毒性的化疗药物5-氟尿嘧啶,并在体外对胶质瘤细胞有显著的生长抑制作用。目的:探讨以骨髓间充质干细胞为基因治疗载体表达外源基因胞嘧啶脱氨酶基因对胶质瘤C6细胞增殖的影响。方法:分离、培养小鼠间充质干细胞,构建胞嘧啶脱氨酶基因与GFP联合的慢病毒载体,通过慢病毒包装法将胞嘧啶脱氨酶基因及GFP转染至小鼠骨髓间充质干细胞,获得稳定表达胞嘧啶脱氨酶基因及GFP的骨髓间充质干细胞,使其与胶质瘤C6细胞共培养,在培养液中加入5-氟胞嘧啶后应用流式细胞仪检测胞嘧啶脱氨酶基因对胶质瘤细胞的增殖影响。结果与结论:慢病毒介导的胞嘧啶脱氨酶基因及GFP基因成功转染小鼠骨髓间充质干细胞形成C57BL/6mMSC-codA/eGFP细胞,C57BL/6mMSC-codA/eGFP在5-氟胞嘧啶的作用下可引起胶质瘤C6细胞的明显凋亡,在5-氟胞嘧啶浓度为1×106μg/L条件下C6胶质瘤细胞凋亡率为60%(P<0.05)。提示,C57BL/6mMSC-codA/eGFP可将5-氟胞嘧啶转化成5-氟尿嘧啶并对C6胶质瘤细胞生长有显著的限制作用甚至是致死效应。更多还原

【Abstract】 BACKGROUND:The bone marrow mesenchymal stem cells(BMSCs) expressing foreign cytosine deaminase gene(gene CD) can effectively transform 5-fluorouracil(5-FC) that was used before chemotherapy into chemotherapeutics 5-FC with cytotoxicity.BMSCs expressing gene CD significantly inhibited the growth of glioma cells in vitro.OBJECTIVE:To explore the effects of BMSCs as gene therapy vector expressing exogenous gene cytosine deaminase gene on the proliferation of glioma C6 cells.METHODS:Mouse MSCs were isolated and cultured.Lentivirus vector combined with cytosine deaminase gene and green fluorescent protein(GFP) was constructed.Using lentivirus packaging,cytosine deaminase gene and GFP were transferred to mouse BMSCs.BMSCs expressing cytosine deaminase gene and GFP were stably obtained and cocultured with glioma C6 cells.Following 5-FC was added to the medium,effects of cytosine deaminase gene on the proliferation of glioma cells were detected using flow cytometry.RESULTS AND CONCLUSION:Lentivirus-mediated cytosine deaminase gene and GFP gene successfully transfected mouse BMSCs and formed C57BL/6 mMSC-codA/eGFP cells.C57BL/6 mMSC-codA/eGFP could induce obvious apoptosis of glioma C6 cells following treatment with 5-FC.The apoptotic rate of C6 glioma cells was 60%(P<0.05) under the action of 5-FC at a concentration of 1×106 μg/L.It was thus concluded that C57BL/6 mMSC-codA/eGFP could convert 5-FC to 5-FU and had significant restriction,even fatal,influence on the growth of C6 glioma cells.更多还原