体外模拟肿瘤微环境中致大鼠MSCs瘤化因子分析

 目的 探讨模拟肿瘤微环境中骨髓间充质干细胞（MSCs）肿瘤转化的发生与HGF、IL-6、BFGF关系。方法 实验组为C6胶质瘤与MSCs间接共培养模拟肿瘤微环境下MSCs的生长，对照组为星型胶质细胞与MSCs间接共培养模拟正常微环境下MSCs的生长，空白对照组MSCs单独培养；QPCR，免疫荧光检测各组的P53和MDM2的基因和蛋白的表达;ELISA检测各组HGF、IL-6、BFGF的表达；免疫荧光检测对应升高的细胞因子作用于MSCs后的STAT3蛋白的表达。结果 实验组的MSCs形态肿瘤化；MSCs的MDM2 mRNA是对照组的1.56±0.21(P<0.05)，MDM2蛋白表达率比对照组升高90±7%(P<0.01)；MSCs的P53的mRNA表达是对照组的0.55±0.10， 而P53突变蛋白表达率比对照组升高87±5%(P<0.01)；HGF、IL-6水平比对照组中达水平升高(P<0.05)；过量IL-6处理组的MSCs的STAT3激活蛋白表达率比正常量IL-6处理组明显升高(P<0.01)。结论 IL-6参与MSCs在肿瘤微环境下的肿瘤转化的发生。

The analysis of the factors which induced rat mesenchymal stem cells tumorigenesis in vitro simulated tumor microenvironment

Objective To investigate the relation between marrow mesenchymal stem cells tumorigenesis and tyrosine kinase signaling molecules such as HGF、IL-6,BFGF in vitro.Methods The tumor microenvironment was simulated by co-culturing MSCs with C6 and the normal microenvironment was simulated by co-culturing MSCs with astrocytes;MSCs were divided into three groups:Experimental group(co-cultured with C6),Control group(co-cultured with astrocytes) Blank control group(MSCs alone);P53 and MDM2 of MSCs in each group were measured by PCR,Western blotting;HGF,IL-6 and BFGF expression in each group were detected by ELISA;the STAT3 protein expression of MSCs treated with corresponding ELISA amount tyrosine kinase signaling molecule were detected by immunofluorescence.Results Experimental group cells showed tumor cell-like morphology.MDM2 mRNA in experimental group was the control group 1.56±0.21 fold(P<0.05);MDM2 protein in experimental group increased 90%±7% as compared to control group(P<0.01);P53 mRNA decreased to 0.55±0.10 and the level of mutant P53 protein increased to 87%±5% in experimental group as compared to that in control group(P<0.01);HGF and IL-6 significantly increased in Experimental group as compared to Control group(P<0.05).P-STAT3 expression increased in IL-6 overdose group as compared to IL-6 normaldose group.(P<0.01).Conclusion IL-6 can induce mesenchymal stem cells tumorigenesis in vitro simulated tumor microenvironment.