Isakari Yoshimasa, Sogo Shinji, Ishida Tatsuhiro, Kawakami Takuma, Ono Toshihide, Taki Takao, Kiwada Hiroshi
Department of Pharmacokinetics and Biopharmaceutics, Subdivision of Biopharmaceutical Sciences, Institute of Health Biosciences, The University of Tokushima, Japan.
Biol Pharm Bull. 2009 Mar;32(3):354-8. doi: 10.1248/bpb.32.354.
A comprehensive gene-expression analysis during platelet (PLT) production from megakaryocytes may give important information on genes involved in the PLT production process. However, the low abundance of primary megakaryocytes makes the gene expression analysis difficult. Therefore, we employed MEG-01 cells, a human megakaryocytic cell line, and confirmed that the cell line produces PLT-like particles by treatment with phorbol myristate acetate (PMA). After treatment of MEG-01 cells with PMA for 8 or 24 h, comprehensive gene expression analysis was carried out using a microarray and Reverse Transcription-Polymerase Chain Reaction (RT-PCR). From the microarray analysis, 141 genes were up-regulated (>2-fold) and 164 genes were down-regulated (<1/2-fold). However, known PLT-related genes were not included in the up- or down-regulated genes. On the other hand, RT-PCR analysis detected increased expression of beta1-tubulin, CD62P, gpIbalpha and gpIII, which are related to PLT function and megakaryocyte differentiation, following PMA treatment for 24 h. These results indicate that the MEG-01 cell may be an alternative model system to study the process of human PLT production from megakaryocytes. The gene-expression analysis might be a powerful tool for identifying genes related to PLT production, if the experimental conditions are optimized.
对巨核细胞生成血小板(PLT)过程进行全面的基因表达分析,可能会提供有关参与PLT生成过程的基因的重要信息。然而,原代巨核细胞数量稀少,使得基因表达分析变得困难。因此,我们使用了人巨核细胞系MEG-01细胞,并证实该细胞系经佛波酯(PMA)处理后可产生类似PLT的颗粒。用PMA处理MEG-01细胞8小时或24小时后,使用微阵列和逆转录聚合酶链反应(RT-PCR)进行全面的基因表达分析。从微阵列分析中,有141个基因上调(>2倍),164个基因下调(<1/2倍)。然而,上调或下调的基因中不包括已知的与PLT相关的基因。另一方面,RT-PCR分析检测到,在PMA处理24小时后,与PLT功能和巨核细胞分化相关的β1-微管蛋白、CD62P、糖蛋白Ibaα和糖蛋白III的表达增加。这些结果表明,MEG-01细胞可能是研究从巨核细胞生成人PLT过程的替代模型系统。如果优化实验条件,基因表达分析可能是识别与PLT生成相关基因的有力工具。