【摘要】 目的通过骨髓间充质干细胞(MSCs)与C6胶质瘤细胞间接共培养,探讨在C6胶质瘤微环境中SIL-6R及GP130的过度激活和表达对MSCs向肿瘤方向转化的影响。方法用Transwell插入式培养皿结合6孔板构建间接共培养体系来模拟MSCs生长的微环境,用RT-QPCR检测细胞中GP130、STAT-3、Cyclin D1和BCL-xl mRNA的表达;相差显微镜下观察细胞形态学;ELISA法检测上清液中SIL-6R的表达;免疫荧光法检测细胞膜上GP130的表达;Western blot检测细胞中STAT-3、Cyclin D1、BCL-xl蛋白表达。结果实验组的MSCs呈现类似胶质瘤细胞样的形态;细胞中GP130、STAT-3、Cyclin D1和BCL-xl mRNA的表达水平和上清液中SIL-6R的表达水平明显高于阴性对照组和空白组(P<0.05);实验组大部分细胞及阳性对照组的细胞膜上表达GP130蛋白,分别达到93%±5%和95%±5%,显著高于阴性对照组及空白组的0%(P<0.01),细胞中STAT-3、Cyclin D1和BCL-xl的蛋白表达水平高于阴性对照组和空白组(P<0.05)。结论 MSCs的恶性转变与SIL-6R/GP130的过度表达和激活存在一定的相关性,但其在MSCs瘤向转变中的作用及机制有待进一步研究。更多还原

【Abstract】 Objective To determine the role of over-expression of SIL-6R and GP130 on oncological differention of bone marrow mesenchymal stem cell(MSCs)by indirectly co-culturing MSCs and C6 glioma cells.Methods The Transwell insert culture dish and 6-well plates were used to build a co-cultured system to simulate the microenvIronment of MSCs growth.GP130 in each group was measured by RT-QPCR and immunofluorescence,STAT-3,Cyclin D1 and BCL-xl in each group was measured by RT-QPCR and Western blot,SIL-6R in each group was measured by ELISA,the cell morphological changes were observed by the phase contrast microscope.Results Experimental group cells showed tumor cell-like morphology.Compared with negative control group and blank group,GP130,STAT-3,Cyclin D1 and BCL-xl mRNA and protein in experimental group were significantly increased(P<0.05),and there was no significant differences between experimental group and positive control group.The expression rate of GP130 reached 93%±5% in experimental group,while in both negative control and blank group was 0%(P<0.05). The expression of SIL-6R in experimental group supernatant was higher than that of negative control and blank control group.Conclusions The oncological differention of MSCs was significantly related to the overexpression and activation of SIL-6R/GP130.But the contribution of SIL-6R/GP130 to the MSCs tumor transformation need to be further confirmed.更多还原