携带hIFN-β的骨髓间充质干细胞体外抑制C6胶质瘤细胞  

C6 glioma cells were inhibited by mesenchymal stem cells(MSCs) transfected with the Beta-interferon gene in vitro

作　　者：田道锋[1] 陈谦学[1] 吴立权[1] 冀保卫[1] 张申起[1] 刘宝辉[1] 王龙[1] 

机构地区：[1]武汉大学人民医院神经外科,430060

出　　处：《卒中与神经疾病》2010年第4期225-228,共4页Stroke and Nervous Diseases

基　　金：国家自然科学基金资助项目(30772223)

摘　　要：目的研究携带人β干扰素基因(hIFN-β)的重组腺病毒转染骨髓间充质干细胞(MSCs-hIFN-β)以及其在体外抑制大鼠C6胶质瘤细胞。方法转染的MSCs行PT-PCR检测细胞内hIFN-βmRNA的表达;ELISA法检测培养液上清hIFN-β的分泌情况;MTT法检测细胞活力并绘制转染后MSCs生长曲线,MSCs-hIFN-β和C6胶质瘤细胞体外共培养,检测C6胶质瘤细胞活力和共培养液上清hIFN-β含量。结果 Ad-hIFN-β转染的MSCs分化后有绿色荧光蛋白表达,RT-PCR证实MSCs中有hIFN-βmRNA表达,ELISA测定显示MSCs分泌hIFN-β,而且转染MSCs的增殖能力无改变。体外共培养发现C6胶质瘤细胞的生长被不同程度的抑制,C6胶质瘤细胞培养上清hIFN-β的含量随着MSCs-hIFN-β的密度增加而增加。结论在体外MSCs-hIFN-β能抑制大鼠C6胶质瘤细胞的增殖。Objective To investigate the inhibition of C6 rat glioma cells by MSCs-IFN-β In vitro.Methods Ad-hIFN-β was transfected into MSCs cultured in vitro.The expression of hIFN-β in MSCs after transfection was determined by observing the expression of hIFN-β and detected by reverse transcription polymerase chain reaction（RT-PCR）.ELISA method was applied to assay the secretion of hIFN-β.MTT method was applied to draw cells growth curve.Cellular viability of C6 glioma cell was assessed by MTT assay and the content of hIFN-β was analyzed using ELISA.Results GFP expression could be observed under fluorescent microscope 24h after infection,and the expression of hIFN-β was confirmed by RT-PCR method.ELISA analysis showed the quantity of hIFN-β was higher than that of control group（P〈0.01）.Moreover,the MSCs growth curve of transfection group and control group had no significant difference（P〉0.0l）.MSCs-hIFN-β significantly inhibited the growth of C6 glioma cell even when the ratio of C6 to MSCs-IFN-β was 100：1.There was a dose-dependent increase in the amount of soluble IFN-β that directly correlated with the inhibition of tumor cell growth.Conclusions C6 glioma cells were inhibited by mesenchymal stem cells（MSCs） transfected with the beta-interferon Gene in vitro.

关 键 词：骨髓间充质干细胞 人β干扰素基因 腺病毒 C6胶质瘤细胞 

分 类 号：R739.4[医药卫生—肿瘤]