Zhao Mingzhi, Xu Feng, Wu Feilin, Yu Debin, Su Na, Zhang Yao, Cheng Long, Xu Ping
State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences Beijing, Beijing Institute of Radiation Medicine , Beijing 102206, P. R. China.
Life Science College, Southwest Forestry University , Kunming 650224, P. R. China.
J Proteome Res. 2016 Dec 2;15(12):4245-4257. doi: 10.1021/acs.jproteome.6b00305. Epub 2016 Nov 10.
HepaRG cell, a stabilized bipotent liver progenitor cell line, exhibits hepatocyte functions only after differentiation. However, the mechanism of transition from nondifferentiated to differentiated states, accompanied by proliferation migration and differentiation, remains poorly understood, particularly those proteins residing in the plasma membrane. In this study, the membrane protein expression change of HepaRG cell during differentiation were systematically analyzed using an iTRAQ labeled quantitative membrane proteomics approach. A total of 70 membrane proteins were identified to be differentially expressed among 849 quantified membrane proteins. Function and disease clustering analysis proved that 11 of these proteins are involved in proliferation, migration, and differentiation. Two key factors (MMP-14 and OCLN) were validated by qRT-PCR and Western blot. Blockade of MMP-14 further demonstrated its important function during tumor cell migration. The data sets have been uploaded to ProteomeXchange with the identifier PXD004752.
HepaRG细胞是一种稳定的双能肝祖细胞系,仅在分化后才表现出肝细胞功能。然而,从未分化状态转变为分化状态并伴有增殖、迁移和分化的机制仍知之甚少,尤其是质膜中的那些蛋白质。在本研究中,使用iTRAQ标记的定量膜蛋白质组学方法系统分析了HepaRG细胞在分化过程中的膜蛋白表达变化。在849种定量膜蛋白中,共鉴定出70种膜蛋白差异表达。功能和疾病聚类分析证明,其中11种蛋白参与增殖、迁移和分化。通过qRT-PCR和蛋白质印迹法验证了两个关键因子(基质金属蛋白酶-14和闭合蛋白)。阻断基质金属蛋白酶-14进一步证明了其在肿瘤细胞迁移过程中的重要作用。数据集已上传至ProteomeXchange,标识符为PXD004752。