Pin1在恶性血液病细胞和细胞周期中的表达

目的:研究Pin1(肽酰脯胺酰顺反式易构酶)在髓系、淋系10种恶性血液病细胞株中的表达情况以及其与不同细胞周期的关系.方法:利用Realtime-PCR检测血液病细胞株的Pin1表达,同时用Thymidine和Nocodazole将细胞阻断在不同的细胞周期,用Realtime-PCR及Western Blot的方法,检测Pin1的mRNA和蛋白的表达水平.结果:Pin1在恶性血液病细胞株中mRNA的表达明显高于正常人骨髓单个核细胞的表达(F=10.508,P＜0.01),其在髓系血液病细胞株和淋系恶性血液病细胞株中的表达均较正常人骨髓单个核细胞明显增高(分别为F=13.584,P＜0.01;F=5.339,P＜0.05).Pin1在G1期mRNA的表达水平显著高于S期(F=40.572,P＜0.05),S期与M期比较没有明显差异(F=8.309,P＞0.05).但在蛋白水平上来看Pin1的变化不明显.结论:Pin1的mRNA在恶性血液病细胞中呈高表达,其表达水平与细胞周期有关,其中以G1期为最高,S期最低.

Expression of Pin1 in malignant hematopoietic cells and its relation with cell cycle

OBJECTIVE: To study the expression of peptidyl-prolyl cis/trans isomerase (PPIase or Pin1) in malignant hematopoietic cells and its relation with cell cycle. METHODS: Realtime quantitative PCR with fluorescence probe hybridization was used to measure expression of Pin1 mRNA in malignant hematopoietic cell lines and normal mononuclear cells separated from bone marrow. HeLa cells were blocked with Thymidine and Nocodazole in different cell phases and then the expression of Pin1 mRNA and protein were detected by realtime-PCR and immunoblotting. RESULTS: The expression of Pin1 in malignant hematopoietic cell lines was significantly higher than that in normal controls (0.339 +/-0.093 compared with 0.038 +/-0.005, P<0.01). Its expression in myeloid malignant hematopoietic cell lines was significantly higher than that in normal controls (0.388 +/-0.115 compared with 0.038 +/-0.005, P<0.01) and so was the malignant lymphocytic cell lines (0.226 +/-0.166 compared with 0.038 +/-0.005, P<0.01). The expression of Pin1 was closely correlated with cell cycle. It was the highest in G1 phase and the lowest in S phase (110.762 +/-16.737 compared with 4.080 +/-0.634, P<0.01). CONCLUSION: Pin1 is overexpressed in malignant hematopoietic cell lines and its expression is different during cell cycle that is highest in G1 phase and lowest in S phase.