【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。更多还原

【Abstract】 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.更多还原