转染人β干扰素基因的骨髓间充质干细胞治疗大鼠颅内胶质瘤

目的 观察转染人β干扰素基因(hIFN-β)的骨髓间充质干细胞(MSCs-hIFN-β)治疗大鼠颅内恶性胶质瘤的作用.方法 重组腺病毒介导hIFN-β基因转染的大鼠骨髓间充质干细胞(MSCs-hIFN-β)和C6胶质瘤细胞体外共培养,噻唑蓝(MTT)比色法检测C6细胞活力,酶联免疫吸附试验(ELISA)检测培养液上清hIFN-β含量;制作SD大鼠颅内胶质瘤模型,并用MRI和组织病理学检查予以证实;MSCs-hIFN-β瘤内注射观察荷瘤鼠的临床表现和生存时间,MRI检查肿瘤的大小,并行大鼠C6胶质瘤组织病理学检查和肿瘤组织hIFN-β免疫组织化学染色.结果 体外共培养发现C6细胞的生长被不同程度地抑制,C6细胞培养上清hIFN-β的含量随着MSCs-hIFN-β的密度增加,其含量也增加;体内实验发现MSCs-hIFN-β、MSCs、生理盐水瘤内注射治疗10 d后肿瘤平均体积分别为(8.43±1.32)、(35.84±2.36)、(37.26±2.91)mm3,治疗组和对照组比较差异有统计学意义(P＜0.01);治疗组较对照组生存期明显延长(P＜0.01).结论 MSCs-hIFN-β能抑制胶质瘤细胞的增殖,延长颅内荷瘤鼠的生存期.

Gene therapy of rat intracranial glioma by Mesenchymal Stem Cells (MSCs) transfected with the beta-interferon gene

Objective To investigate the gene therapy of experimental rat intracranial glioma by MSCs-hIFN-β. Methods Cellular viability of C6 glioma cell was assessed by MTT assay and the content of hIFN-β was analyzed using enzyme linked immunosorbent assay (ELISA). C6 glioma model was established and was verified by histopathological characteristics and MRI images. Clinical manifestation of the rat bearing C6 glioma were recorded. Direct intratumoral injection of MSCs-IFN-β gene was undertaken, survival of mices were recorded. The expression of protein of hIFN-β in C6 glioma tissure was examined immunohistochemically. Results MSCs transfected with IFN-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. There was significant difference between the mean tumor volume of group MSCs-IFN-β and group saline, MSCs after 10 days. Compared with animals treated with saline or with MSCs, treated with MSCs-IFN-β resulted in a significant increase in animal survival (P ＜ 0. 0l ). Conclusion In vitro and in vivo efficacy studies show that MSCs can be engineered to release IFN-β and that these engineered MSCs can be exploited to therapeutic advantage against gliomas.