Proteomic Profiling of Human Hepatic Stellate Cell Line LX2 Responses to Irradiation and TGF-beta 1
Proteomic Profiling of Human Hepatic Stellate Cell Line LX2 Responses to Irradiation and TGF-beta 1
复制标题
人肝星状细胞系 LX2 对辐射和 TGF-β1 反应的蛋白质组学分析
DOI：
10.1021/acs.jproteome.8b00814
复制
发表时间：
2019
影响因子：
4.4
通讯作者：
Zeng Zhaochong
中科院分区：
文献类型：
--
作者：
Yuan Baoying;Chen Yuhan;Wu Zhifeng;Zhang Li;Zhuang Yuan;Zhao Xiaomei;Niu Hao;Cheng Jason Chia-Hsien;Zeng Zhaochong
肝星状细胞（HSC）是辐射损伤的主要靶细胞，主要参与辐射诱导的肝纤维化的发展。然而，辐射诱导的HSC激活的分子事件尚未完全阐明。在本研究中，用X射线照射和/或TGF-β1处理人HSC系LX 2，并评估促纤维化分子。进行iTRAQ LC-MS/MS技术以鉴定暴露于不同刺激后LX 2中的总体蛋白质表达谱。照射或TGF-β1单独作用可增加α-SMA、胶原1、CTGF、派-1和纤维连接蛋白的表达。辐射和TGF-β1协同诱导这些促纤维化标志物的表达。在受照射、TGF-β1或共处理影响的LX 2细胞样品中，总共分别鉴定了102、137、155个失调蛋白。生物信息学分析表明，这三组差异表达蛋白通常与细胞周期和内质网蛋白质加工有关。一组蛋白质的表达得到了适当的验证：照射或照射和TGF-β1共刺激后，CDC 20，PRC 1，KIF 20 A，CCNB 1，SHCBP，TACC 3上调，而TGF-β1或TGF-β1加照射处理后，THBS 1和THBS 1升高。此外，CDC 20抑制抑制了照射和TGF-β1刺激的LX 2细胞中促纤维化标志物的表达。本文所提供的关于引起辐射诱导的HSC活化的潜在分子机制的详细数据将有助于制定最大限度地减少辐射诱导的肝纤维化的放射治疗策略。
Hepatic stellate cells (HSCs) are the main target of radiation damage and primarily contribute to the development of radiation-induced liver fibrosis. However, the molecular events underlying the radiation-induced activation of HSCs are not fully elucidated. In the present study, human HSC line LX2 was treated with X-ray irradiation and/or TGF-β1, and profibrogenic molecules were evaluated. The iTRAQ LC–MS/MS technology was performed to identify global protein expression profiles in LX2 following exposure to different stimuli. Irradiation or TGF-β1 alone increased expression of α-SMA, collagen 1, CTGF, PAI-1, and fibronectin. Irradiation and TGF-β1 cooperatively induced expression of these profibrotic markers. In total, 102, 137, 155 dysregulated proteins were identified in LX2 cell samples affected by irradiation, TGF-β1, or cotreatment, respectively. Bioinformatic analyses showed that the three differentially expressed protein sets were commonly associated with cell cycle and protein processing in endoplasmic reticulum. The expression of a set of proteins was properly validated: CDC20, PRC1, KIF20A, CCNB1, SHCBP, TACC3 were upregulated upon irradiation or irradiation and TGF-β1 costimulation, whereas SPARC and THBS1 were elevated by TGF-β1 or TGF-β1 plus irradiation treatment. Furthermore, CDC20 inhibition suppressed expression of profibrotic markers in irradiated and TGF-β1-stimulated LX2 cells. Detailed data on potential molecular mechanisms causing the radiation-induced HSC activation presented here would be instrumental in developing radiotherapy strategies that minimize radiation-induced liver fibrosis.