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目的:为了对小鼠H22肝癌细胞中高表达膜蛋白进行初步的分析并探索其与肿瘤细胞恶性行为的相关关系。方法:分别培养并裂解小鼠正常肝细胞及H22肝癌细胞株,采用疏水性裂解液分步提取膜蛋白,双向电泳分离。ImageMaster 2DPlatinum软件对电泳图谱进行对比分析,选择在H22细胞中典型的高表达膜蛋白并采用MALDI-TOF-MS进行鉴定。结果:从H22细胞中提取的膜蛋白约60%表现为高表达。8个典型的在H22细胞中高表达的膜蛋白被鉴定为紧联蛋白ZO-2、HMG-CoA还原酶、囊泡蛋白分选相关蛋白52、分选和装配结构组分50、巨噬细胞清道夫受体Ⅰ/Ⅱ型、硫酸酯酶修饰因子2、PKC和CKⅡ底物蛋白3和C9orf135蛋白。结论:H22肝癌细胞中大多数膜蛋白表现为高表达。从已鉴定的膜蛋白推测,H22细胞功能改变可能涉及细胞迁移、膜融合、信号转导、基因表达调控及能量代谢等,这种改变可能与肿瘤细胞的恶性行为相关。
OBJECTIVE: To analyze the high expression of membrane proteins in mouse H22 hepatocellular carcinoma cells and to explore their relationship with the malignant behavior of tumor cells. Methods: The normal hepatocytes and H22 hepatocarcinoma cell lines were cultured and lysed respectively. The membrane proteins were extracted by hydrophobic lysis solution and separated by two-dimensional electrophoresis. ImageMaster 2DPlatinum software for comparative analysis of electrophoresis, select the typical high expression of membrane protein in H22 cells and identified by MALDI-TOF-MS. Results: About 60% of the membrane proteins extracted from H22 cells showed high expression. Eight typical membrane proteins that are highly expressed in H22 cells were identified as the tight junction protein ZO-2, HMG-CoA reductase, vesicle sorting related protein 52, sorting and assembly of structural components 50, Receptor type I / II, sulfatase modifier 2, PKC and CKII substrate protein 3 and C9orf135 protein. Conclusion: Most membrane proteins in H22 hepatoma cells are highly expressed. It is speculated that the functional changes of H22 cells may involve cell migration, membrane fusion, signal transduction, gene expression regulation and energy metabolism, which may be related to the malignant behavior of tumor cells.