目的 探讨PEX基因对C6胶质瘤干细胞(GSCs)中基质金属蛋白酶2(MMP-2)和血管内皮生长因子(VEGF)表达的影响.方法 采用血清培养基培养C6胶质瘤细胞系,再取其单细胞悬液悬浮培养于无血清培养基中传代培养.采用单细胞克隆和免疫荧光染色分离鉴定GSCs,运用本实验室成功构建的重组子pDsRed2-C1-PEX,以脂质体为介导转染体外培养的GSCs,逆转录酶-聚合酶链反应(RT-PCR)检验真核表达载体在GSCs中的表达及PEX基因对GSCs表达MMP-2mRNA、VEGFmNA的影响.结果 成功培养出GSCs并成功转染,PEX在GSCs中高表达并且体外对MMP-2、VEGF的表达有明显抑制作用,差异有统计学意义(P＜0.05).结论 PEX基因可以在GSCs中表达,并且体外可以抑制MMP-2和VEGFmRNA在GSCs中的表达,为胶质瘤的治疗奠定理论基础.%Objective To study the effects of PEX gene on expression of matrix metalloproteinase 2 ( MMP-2) and vascular endothelial growth factor ( VEGF) of C6 glioma cells. Methods C6 cell lines were cultured in traditional serum medium culture and the single-celled levitation liquid suspension was cultured in serum-free. Glioma stem cells were separated and identified by single-celled cloning and immune fluorescence staining and the pDsRed2-Cl-PEX which was successfully constructed was used to transfect glioma stem cells (GSCs) in vitro mediated by liposome. Expression of GCSs and effect of PEX gene on the expression of MMP-2 and VEGF mRNA were detected by using reverse transcriptase-polymerase chain reaction ( RT-PCR). Results GSCs were successfully cultured and transfected. PEX was highly expressed in GSCs and have inhibitory effect on the expressions of MMP-2 and VEGF of G5 glioma stem cells in vitro with significant differences (P < 0. 05). Conclusion PEX gene can be expressed in C6 glioma stem cells, and it can suppress the expressions of MMP-2 and VEGF mRNA in GSCs in vitro, which lay a theoretical foundation for the treatment of glioma.
展开▼
