转染pCE-GFP的重组骨髓间充质干细胞对Walker-256细胞的体外抗肿瘤作用

Anti-tumor Effects of BMSCs Transfected with pCE-GFP on Walker-256 Cells

目的转染pCE-GFP质粒至大鼠的骨髓间充质干细胞(BMSCs),研究重组BMSCs(rBMSCs)对Walker-256细胞的体外抗肿瘤作用。方法贴壁培养法分离获得BMSCs,Lipofectamine 2000转染pCE-GFP质粒至BMSCs,rBM-SCs经肝细胞生长因子(hepatocyte growth factor,HGF)处理促进分化。采用ELISA法检测rBMSCs培养上清中白蛋白和甲胎蛋白水平;MTT法检测rBMSCs培养上清对Walker-256细胞生长增殖的影响;RT-PCR法检测rBMSCs培养上清对Walker-256细胞Bcl-2和Bax mRNA表达的影响;采用细胞共培养跨膜迁移实验,观察rBMSCs趋化能力的改变。结果 rBMSCs培养上清对大鼠Walker-256细胞增殖抑制作用较BMSCs强,差异有统计学意义(P<0.05);经HGF处理的rBMSCs组作用强于未经HGF处理的rBMSCs组,两组比较差异有统计学意义(P<0.05)。经HGF处理的rBM-SCs组与其他3组比较,Bax mRNA表达显著上调(P<0.05);Bcl-2 mRNA表达在各组中未见明显改变。rBMSCs的趋化能力增强,与BMSCs组比较差异有统计学意义(P<0.05),且经HGF处理的rBMSCs组趋化能力强于未经HGF处理的rBMSCs组(P<0.05)。结论经HGF处理的rBMSCs通过调控Walker-256细胞的增殖和凋亡作用及增强对肿瘤细胞的趋化能力而发挥抗肿瘤作用。

Objective To study the anti-tumor effects of bone marrow mesenchymal stem cells（BMSCs）transfected with pCE-GFP plasmid（rBMSCs）on Walker-256 cells.Methods BMSCs were isolated and cultured,transfected with pCE-GFP plasmid by lipofectamine 2000,treated by hepatocyte growth factor（HFG）,and observed under the microscopy for cell morphology.ELISA was used to determine the levels of albumin and AFP protein in the culture supernatant of rBMSCs.MTT assay was applied to measure the effects of the culture supernatant of rBMSCs on growth and proliferation of Walker-256 cells.By using RT-PCR,the expression of Bcl-2 and Bax mRNA was detected in Walker-256 cells.By using Transwell assay,the chemotactic ability of BMSCs was observed.Results The culture supernatant of rBMSCs in each group could inhibit the growth of Walker-256 cells more significantly than that of BMSCs（P0.05）,and more significantly in rBMSCs treated with HGF than in those treated without HGF（P0.05）.In rBMSCs treated with HGF,the expression of Bax mRNA was significantly up-regulated as compared with other three groups（P0.05）,but there was no significant difference in the expression of Bcl-2 mRNA among four groups（P0.05）.The chemotactic ability of rBMSCs was significantly enhanced as compared with BMSCs,and that of rBMSCs treated with HFG was significantly increased as compared with rBMSCs not treated with HFG（P0.05）.Conclusion rBMSCs treated with HFG exert their antitumor effects by modulating proliferation and apoptosis of Walker-256 cells,and promoting the chemotactic ability.