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旰细胞及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 2D Platinum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CKU底物蛋白3和C90rf135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细胞功能改蛮可能涉及细胞迁移、膜融合、信号转导、基因表达调摔及能量代谢等.这种改变可能与肿瘤细胞的恶性行为相关。
Objective: To preliminarily analyze high expressed membrane proteins in mouse hepatoeellular carcinoma cell line H22 and explore their relationship with malignant behavior of tumor cells. Methods: Mouse normal liver cells and hepatocarcinoma cell line H22 were cultured and lysed respectively. The membrane proteins were extracted step by step with hydrophobic lysis solution and separated by two-dimensional electrophoresis (2-DE). The differential proteins of the 2-DE maps were matched and analyzed by ImageMaster 2D Platinum software. Then the membrane proteins with high expression in H22 cells were selected and identified by MALDI- TOF-MS. Results: The eight differential membrane proteins with high expression of H22 cells were identified as ZO-2, HMG-CoA reductase, VPS52, SAM50, MSR types Ⅰ/Ⅱ, SUMF2, PACSIN3 and C9orf135 protein. Conclu- sion: Most of membrane proteins in H22 cells show a high expression. Inferring from identified membrane pro- teins, the functional alterations in H22 cells involve cellular transmigration, membrane fusion, signal transduction, gene expression regulation, energy metabolism and so on, and these changes may be related with malignant behavior of tumor cells.