白细胞介素18、单纯疱疹病毒-胸苷激酶基因联合修饰骨髓间充质干细胞治疗脑胶质瘤  

Interleukin-18-and HSV-tk-expressing bone marrow stromal cells exhibit antiglioma activity.

作　　者：李春晖[1] 焦保华[2] 康春生[3] 史彦芳[1] 郭毅[1] 崔凯[1] 

机构地区：[1]河北大学附属医院神经外科,保定071000 [2]河北医科大学第二医院神经外科 [3]天津医科大学总医院神经外科

出　　处：《中国神经精神疾病杂志》2010年第6期325-330,共6页Chinese Journal of Nervous and Mental Diseases

摘　　要：目的了解骨髓间充质干细胞(bone marrow stromal cells,BMSCs)联合转染单纯疱疹病毒-胸苷激酶基因(HSV-tk)和白细胞介素18(interleukin-18,IL-18)后在荷瘤大鼠体内的治疗作用。方法提取1周龄大鼠骨髓培养大鼠骨髓间充质干细胞。骨髓间充质干细胞转染单纯疱疹病毒-胸苷激酶基因(BMSCs/tk)或白细胞介素18(BMSCs/IL-18)或联合转染单纯疱疹病毒-胸苷激酶基因、白细胞介素18(BMSCs/tk/IL-18),而后进行体内外试验检测其抗肿瘤作用。检测荷瘤鼠脾脏淋巴细胞免疫活性,采用TUNEL方法检测胶质瘤内的凋亡细胞,抗-CD34染色检测微血管密度(MVD),检测淋巴细胞的浸润,记录试验大鼠的生存期。结果骨髓间充质干细胞可稳定转染IL-18及tk基因,BMSCs/tk和BMSCs/IL-18/tk均显示了明显的旁观者效应,荷瘤鼠移植BMSCs/IL-18或BMSCs/IL-18/tk后,血清白细胞介素2和干扰素-γ浓度明显增高,并且接受移植的荷瘤鼠再次受到C6细胞冲击后可诱导快速的抗肿瘤免疫反应。微血管密度检测显示,BMSCs/IL-18(4.50±2.19)、BMSCs/tk/IL-18(3.65±2.13)与对照组(7.15±2.11)、PBS组(7.55±1.64)、BMSCs组(7.80±2.28)、BMSCs/tk组(7.65±1.84)比较有明显低的微血管密度(P<0.05),通过TUNEL检测、浸润淋巴细胞计数、细胞因子浓度及荷瘤鼠生存期比较,BMSCs/IL-18/tk比BMSCs/IL-18或BMSCs/tk显示了更加明显的抗肿瘤作用,BMSCs/IL-18/tk组生存期明显延长。结论联合转染单纯疱疹病毒-胸苷激酶基因和白细胞介素18的骨髓间充质干细胞显示了明显的抗恶性胶质瘤作用,骨髓间充质干细胞可能成为一种理想的治疗颅内胶质瘤的基因治疗载体。Objective To investigate the effects of bone marrow stromal cells（BMSCs） co-transfected with IL-18-and HSV-TK on glioma-bearing rats.Methods BMSCs were isolated from one week old rats and were transfected with HSV-tk（BMSCs/tk） or rmIL-18（BMSCs/IL-18） or both HSV-tk and rmIL-18（BMSCs/tk/IL-18）.Spleen lymphocyte was isolated to detect their immunity activity.TUNEL was used to identify apoptotic cells inside glioma.Anti-CD34,anti-CD8 and anti-CD4 staining was performed to evaluate microvessel density（MVD） and lymphocyte infiltration.The survival times of all rats were recorded.Results BMSCs co-transfected with IL-18 and HSV-tk could express IL-18 and HSV-tk stably.Both BMSCs/tk and BMSCs/IL-18/tk exhibited an obvious bystander effect.Transplantation with BMSCs/IL-18 or BMSCs/IL-18/tk increased the serum concentrations of IL-2 and IFN-γ in glioma-bearing rats and those rats exhibited a rapid immune response when rechallenged with C6 cells.The antiglioma activity of BMSCs/IL-18/tk was stronger compared with BMSCs/IL-18 or BMSCs/tk.The microvessel density was 7.15±2.11,,7.55±1.64,7.80±2.28 and 7.65±1.84,4.50±2.19 and 3.65±2.13 in control,PBS,BMSCs,BMSCs/tk,BMSCs/IL-18 and BMSCs/tk/IL-18 groups,respectively.The microvessel density was lower in BMSCs/IL-18 and BMSCs/tk/IL-18 groups.Conclusions BMSCs co-transfected with IL-18-and HSV-TK has a strong antiglioma activity and thus has a clinical implication in treatment of malignant gliomas.

关 键 词：骨髓间充质干细胞 白细胞介素18 胶质瘤 单纯疱疹病毒-胸苷激酶基因 

分 类 号：R739.41[医药卫生—肿瘤]