转染HSV-tk骨髓基质干细胞旁观者效应机制的研究  

Study on mechanism of bystander effect induced by BMSCs transfected with HSV-tk

作　　者：李春晖[1] 焦保华[1] 

机构地区：[1]河北医科大学附属第二医院神经外科,河北石家庄050000

出　　处：《中华肿瘤防治杂志》2007年第23期1778-1782,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：河北省自然科学基金项目(C2004000559)

摘　　要：目的:研究转染单纯疱疹病毒-胸苷激酶(HSV-tk)基因的骨髓基质干细胞(BMSCs)旁观者效应的机制。方法:培养BMSCs,扩增Ad-CMV-tk后转染骨髓基质干细胞(BMSCs/tk);应用Transwell培养板使BMSCs/tk与C6细胞不相互接触,观察旁观者效应;观察0.22μm滤膜过滤后的条件培养液对C6细胞的作用。建立大鼠脑荷瘤模型,移植BMSCs/tk后,测定其血清中IFN-γ的变化;免疫组化观察荷瘤大鼠肿瘤内部淋巴细胞浸润及微血管密度。结果:在混合培养实验中,BMSCs/HSV-tk诱导BM-SCs的凋亡率明显高于对C6的凋亡率,两者差异有统计学意义,q=8.214,P=0.032。应用Transwell培养板培养后,BMSCs/HSV-tk诱导BMSCs凋亡率明显降低。BMSCs/HSV-tk/G-CV培养上清液可明显诱导C6细胞凋亡,但将其用0.22μm的滤膜过滤后,诱导凋亡的作用消失。脑荷瘤大鼠移植BMSCs/HSV-tk后血清IFN-γ浓度明显增高,肿瘤组织内CD4+细胞数、CD8+细胞数明显增加,微血管密度降低。结论:BMSCs/HSV-tk通过诱导BMSCs细胞凋亡后释放的凋亡小体介导旁观者效应,并发挥免疫学调节机制,对C6细胞可产生明显的抑制作用。OBJECTIVE： To study the mechanism of bystander effect induced by BMSCs transfected with HSV-tk. METHODS： BMSCs were cultured and transfected with AdCMV-tk （BMSCs/tk）, and then, for observing bystander effect, BMSCs/tk were co-cultured with C6 cells in Transwell plate to prevent mixture. After filtrated with 0. 22 μm membrane or not, the selective culture medium of BMSCs/tk after treated with GCV was added into the culture plate of C6 cells. After glioma-bearing rats were established and BMSCs/tk transplantation was performed, IFN-γ concentrations in serum were measured, and lymphocytes infiltrated and microvessel density in glioma were observed by means of immunohistochemistry. RESULTS： When BMSCs/HSV-tk were mixed with BMSCs, the obvious multiplication suppression was observed, but when BMSCs/HSV-tk were mixed with C6 cells, there was fewer multiplication suppression, q = 8. 214, P 0. 032. When co-cultured in Transwell plate, the multiplication suppression was decreased. The selective culture medium of BMSCs/HSV-tk/GCV had an obvious effect to suppress the cell multiplication, but when filtrated with 0. 22 μm membrane, its effect disappeared. After transplantation of BMSCs/tk, the IFN-γ serum concentration of glioma bearing rats was increased, infiltrated CD4^＋ lymphocytes and CD8^＋ lymphocytes were increased obviously, and the microvessel density was decreased. CONCLUSION： BMSCs/HSV-tk can suppress C6 cells obviously through the apoptosis body which is released by BMSCs and the immunity effect is induced.

关 键 词：骨髓细胞/细胞学 间质干细胞 单纯疱疹病毒属 胞苷激酶 旁观者效应 

分 类 号：R739.4[医药卫生—肿瘤]