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目的探讨星形胶质瘤细胞与星形胶质细胞间的蛋白谱变化。方法选择星形细胞来源的 C6胶质瘤细胞和体外培养纯化大鼠皮层的星形胶质细胞进行蛋白组学分析。星形细胞和 C6细胞双向凝胶电泳(2-DE)的凝胶图像用 PDQuest 软件比较,差异蛋白质点进行基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)和生物信息分析,部分蛋白质的差异表达结果用半定量逆转录聚合酶链反应(RT-PCR)验证。结果获得了24个有明显表达变化的蛋白点;17个蛋白质得到质谱结果,与星形胶质细胞相比,经生物信息学分析确认的6个蛋白质中,磷酸甘油酸变位酶1、生物转化酶、谷胱甘肽 S-转移酶 P、钙网织蛋白和膜联蛋白 A2在 C6细胞中表达量较高,而波形蛋白和肌动蛋白γ-肌动蛋白在 C6细胞中表达量较低。磷酸甘油酸变位酶1、膜联蛋白 A2和波形蛋白经半定量RT-PCR 验证与2-DE 结果相符。结论本研究发现的差异表达蛋白质与许多重要的细胞生理活动和功能有密切关系,包括无氧酵解过程、细胞骨架组装、生物转化和信号转导,可能与胶质瘤的生长迅速、浸润迁移和抗药性相关。
Objective To investigate the changes of protein profile between astrocytes and astrocytes. Methods Astrocytes derived from astrocyte-derived C6 glioma cells were cultured and purified in vitro for proteomics analysis. Gel images of astrocytes and C6 cells by two-dimensional gel electrophoresis (2-DE) were compared using PDQuest software, and protein spots were analyzed by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF- MS) and bioinformatics analysis. The differential expression results were verified by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Results 24 protein spots with obvious expression changes were obtained. 17 proteins obtained MS results. Compared with astrocytes, among the 6 proteins confirmed by bioinformatics analysis, phosphoglycerate mutase 1, Invertase, glutathione S-transferase P, calcium reticulin and annexin A2 were higher in C6 cells than those in C6 cells, while the expression of vimentin and actin gamma-actin in C6 cells was higher low. Phosphoglycerate mutase 1, annexin A2 and vimentin were verified by semi-quantitative RT-PCR and 2-DE results. Conclusions The differentially expressed proteins found in this study are closely related to many important cellular physiological activities and functions including anaerobic glycolysis, cytoskeletal assembly, biotransformation and signal transduction, which may be associated with the rapid growth of gliomas, infiltration and migration And resistance related.