肝癌细胞株HepG2的G1期、G2/M期细胞蛋白质组表达差异的研究  

Research of Total Protome Differential Expression in Hepatocellular Carcinoma Cell HepG2 between G1 Phase and G2/M phase

作　　者：白瑞霞[1] 王文礼[1] 

机构地区：[1]内蒙古医学院,呼和浩特010059

出　　处：《科学技术与工程》2009年第3期572-580,共9页Science Technology and Engineering

基　　金：内蒙古医学院重大科研项目(ny2004-zd-002)资助

摘　　要：通过TdR(胸腺嘧啶核苷)调控人肝癌细胞株HepG2的细胞周期,运用双向电泳-图像分析-质谱技术研究肝癌细胞株HepG2的G1期、G2/M期全细胞蛋白质组表达的差异,探索参与肝癌细胞株HepG2的G1期、G2/M期调控相关的蛋白。以TdR诱导肝癌细胞株HepG2,分别得到同步化的G1期、G2/M期细胞,流式细胞术检测。采用比较蛋白质组学的研究技术(双向电泳、质谱分析)筛选G1期、G2/M期差异表达的蛋白质。结果流式细胞术检测显示TdR诱导后,分别获得(63.62±2.82)%的G2/M期同步化细胞,(75.24±0.17)%的G1期同步化细胞,通过双向电泳-图像分析-质谱技术得到21个差异表达的蛋白。其中G2/M期表达,G1期未见表达有10种蛋白;存在三倍以上的差异点11个:2种在G2期表达上调,9种在G1期表达上调。说明TdR阻断法能够获得很好的同步化细胞。质谱鉴定得到的这些差异表达的蛋白质涉及到细胞周期调控、DNA结合、转录调控、mRNA剪接、肽链合成起始、折叠以及细胞代谢等方面。To control cell cycle of human hepatocellular carcinoma cell of line HepG2 by thymidine （TdR） and research of total proteome differential expression in HepG2 between G1 phase and G2/M phase using technology of two-dimensional electrophoresis-mass spectrum, in order to explore some cell cycle related proteins, cultured cells were blocked by TdR . Cells in G1 phase and G2/M phase were lysated Then assayed by flow cytometry （FCM）. The differential expressed proteins in G1 phase and G2/M phase were screened using different proteomics technique （two-dimensional electrophoresis and mass spectrum）. It is obtained FCM showed that after beeing induced by TdR, （63.62 ＋2.82）% and （75.24 ＋0.17）% synchronous cancer cells in G1 phase and G2/M phase were achieved respectively. 21 differential expressed proteins were achieved via technique of Two-dimensional electrophoresis image analysis mass spectrum. Among them 10 were expressed both in G2/M phase and the end of G1 phase . There were over triplication difference among 11 differential spots . two of them were upregulated expressed in G2 phase and nine were upregulated expressed in G2/M phase. It is conclused that TdR can achieve synchronous cancer cells very well. The differential expressed proteins identified by mass spectrum involved in many aspects, for example , regulation of cell cycle, DNA binding, transcriptional control, mRNA splaing, onset of polypeptide chain synthesis ,folded as well as cell metabolism and so on.

关 键 词：肝癌 细胞周期 蛋白质组学 

分 类 号：R333.4[医药卫生—人体生理学]