转染IL-18的骨髓基质干细胞对大鼠脑胶质瘤的作用  

Effect of Bone Marrow Stromal Cells Transfected with Interleukin 18 on Growth of Intracranial Glioma in Rats

作　　者：李春晖[1] 焦保华[1] 

机构地区：[1]河北医科大学第二附属医院神经外科,河北石家庄050000

出　　处：《癌症》2007年第1期38-43,共6页Chinese Journal of Cancer

基　　金：河北省自然科学基金(No.C2004000559)~~

摘　　要：背景与目的:脑胶质瘤因其广泛的浸润性、免疫逃逸等生物学特性使手术、放疗、化疗等方法疗效不佳。本实验观察具有肿瘤趋向性的骨髓基质干细胞(bonemarrowstromalcells,BMSCs)转染IL-18后对大鼠脑胶质瘤的作用。方法:贴壁法培养大鼠骨髓细胞获取纯化的BMSCs,采用流式细胞术鉴定。逆转录病毒LXSN/IL-18转染BMSCs,RT-PCR、免疫荧光检测IL-18基因转录及表达情况,MTT、流式细胞术检测转染前后BMSCs细胞部分生物学特性的变化。ELISA检测BMSCs/IL-18培养上清液对大鼠淋巴细胞的激活作用。体内实验中将荷瘤鼠模型分为4组,实验Ⅰ组、实验Ⅱ组、PBS组大鼠中分别注入BMSCs细胞、BMSCs/IL-18细胞和PBS,对照组不作处理,检测BMSCs/IL-18对荷瘤鼠的作用。结果:BMSCs/IL-18可稳定转录及表达IL-18,但转染后细胞增殖速度减慢,其培养上清液可诱导大鼠淋巴细胞IFN-γ分泌量增长约9倍。脑磁共振检测显示各组肿瘤体积为:实验组Ⅰ(18.26±6.84)mm3,实验组Ⅱ(6.37±1.52)mm3,PBS组(22.48±6.02)mm3,对照组(21.06±5.83)mm3。各组大鼠存活期分别为:实验组Ⅰ(25.3±6.4)天,实验组Ⅱ(84.7±16.3)天,PBS组(21.6±4.7)天,对照组(22.5±6.2)天。荷瘤大鼠脑内移植BMSCs/IL-18后,肿瘤组织中淋巴细胞数量增多,每个200倍光镜视野中,CD4+淋巴细胞数为(37.7±3.5)个,CD8+淋巴细胞数为(32.3±4.5)个。结论:BMSCs可稳定表达转染基因,BMSCs/IL-18对大鼠胶质瘤的生长有明显抑制作用。BACKGROUND ＆ OBJECTIVE： Because of the invasion mmune escape characteristics, surgical resection, radiotherapy, and and chemotherapy had no definite curative effects on intracranial glioma. Because bone marrow stromal cells （BMSCs） can track migrating cells, and interleukin 18 （IL-18） can enhance antitumor immune reaction, this study was to explore the effect of IL-18-transfected BMSCs on growth of glioma in rats. METHODS. Pure BMSCs were obtained by culturing rat bone marrow cells and identified by flow cytometry （FCM）. BMSCs were transfected with retrovirus LXSN/IL-18 to prepare BMSCs/IL-18. IL-18 genetic transcription and expression were assessed by reverse transcription-polymerase chain reaction （RT-PCR） and immunofluorescence. The characteristic variations of BMSCs were detected by MTT assay, FCM, and immunofluorescence. The effect of BMSCs/IL-18 on activation of T cells was evaluated by ELISA. Glioma-bearing rats were divided into BMSCs group, BMSCs/IL-18 group, PBS group, and control group and received relevant treatments. The effect of BMSCs/IL-18 on growth of glioma was observed. RESULTS： IL-18 gene was expressed stably in BMSCs/IL-18. The proliferation speed of BMSCs/IL- 18 was slower than that of BMSCs. The secretion of interferon-γ （IFN-γ） from rat spleen lymphocytes in BMSCs/IL-18 group was 9 times more than that in BMSCs group. Tumor volume was （18.26±6.84） mm^3 in BMSCs group, （6.37±1.52） mm^3 in BMSCs/IL-18 group, （22.48±6.02） mm^3 in PBS group, and （21.06±5.83） mm^3 in control group; survival time of the rats was （25.3±6.4） days, （84.7±16.3） days, （21.6±4.7） days, and （22.5±6.2） days, respectively. After transplantation of BMSCs/IL-18, CD4^＋ T cells in glioma were increased to 37.7±3.5 and CD8^＋ T cells were increased to 32.3±4.5 in each field of view （×200）. CONCLUSION： BMSCs/IL-18 could express IL- 18 gene stably, and have definite therapeutic effect on glioma in rats.

关 键 词：胶质瘤 IL-18 骨髓基质干细胞 干扰素-Γ 大鼠 

分 类 号：R739.41[医药卫生—肿瘤]