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该研究探讨了骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)在C6脑胶质瘤细胞模拟的肿瘤微环境中恶性转化的规避方法,为BMSCs的安全应用提供参考依据。实验分为三组:BMSCs与C6脑胶质瘤细胞间接共培养未处理组、间接共培养+DMSO处理组和间接共培养+TPCA1(3-thiophenecarboxamide,2-[(aminocarbony 1)amino]-5-(4-fluoropheny 1)1)处理组。显微镜下观察各组细胞的形态变化;Transwell实验检测各组细胞的迁移和侵袭能力;Real-time PCR检测各组细胞STAT3(signal transducers and activators of transcription 3)、NF-κB(nuclear factor-κB)/p56和C-myc m RNA水平;Western blot和免疫荧光检测各组细胞P-STAT3、NF-κB/p56和C-myc蛋白质水平。结果表明,实验组中间接共培养+TPCA1处理组细胞的规整性明显比间接共培养未处理组好;间接共培养+TPCA1处理组细胞迁移和侵袭能力明显低于未处理组;间接共培养+TPCA1处理组细胞STAT3、NF-κB/p56和C-myc m RNA和蛋白质水平明显低于未处理组(P<0.05)。该研究结果表明,TPCA1能改变处于C6脑胶质瘤细胞模拟的瘤肿瘤微环境中的BMSCs的生物学特性。
This study explored the ways to avoid the malignant transformation of bone marrow mesenchymal stem cells (BMSCs) in the tumor microenvironment simulated by C6 glioma cells, and provided a reference for the safe application of BMSCs. The experiment was divided into three groups: Indirect co-culture of BMSCs and C6 glioma untreated group, Indirect co-culture + DMSO treatment group and indirect co-culture + TPCA1 (3 - thiophenecarboxamide, - (4-fluoropheny 1) 1) treatment group. The morphological changes of the cells in each group were observed under the microscope; the migration and invasion ability of the cells in each group were detected by Transwell assay; the expressions of STAT3 (signal transducers and activators of transcription 3), NF-κB / p56 and C-myc m RNA levels; Western blot and immunofluorescence were used to detect the protein levels of P-STAT3, NF-κB / p56 and C-myc in each group. The results showed that the regularity of TCCA1 + TPCA1-treated cells in the experimental group was significantly better than those in the indirect co-culture and untransfected cells. The ability of cell migration and invasion in indirect co-cultured + TPCA1-treated cells was significantly lower than that in the untreated group. The levels of STAT3, NF-κB / p56 and C-myc mRNA and protein in TPCA1-treated cells were significantly lower than those in untreated cells (P <0.05). The results of this study indicate that TPCA1 alters the biological properties of BMSCs in a tumor tumor microenvironment that is modeled by C6 glioma cells.