Xue Yunping, Xu Pengfei, Xu Sujuan, Xue Kai, Xu Lingling, Chen Jie, Xu Juan, Shi Xiaoyan, Li Qian, Gu Lin
Nanjing Maternal and Child Health Institute, Nanjing, China.
Department of Gynecological Endocrinology, Nanjing, China.
Cell Physiol Biochem. 2018;47(1):107-118. doi: 10.1159/000489753. Epub 2018 May 9.
BACKGROUND/AIMS: Ovarian endometriosis (OvE) is ovarian cyst that is lined with endometrial tissue. They are found in 17-44% of women with endometriosis. Their clinical manifestations include pelvic pain, dysmenorrhea, dyspareunia, and infertility. Although the incidence of OvE has increased yearly, the exact pathogenesis of OvE is still unclear. We used peptidomics, an emerging branch of proteomics, to identify differentially expressed peptides in order to determine the possible roles of these peptides in the pathogenesis of OvE.
The ectopic and eutopic endometria of OvE were used to extract peptides with 10-kDa molecular weight cutoff filters, and the peptide precursor proteins were then identified with PEAKS software, followed by quantification with the TMT labeling method and subsequent analysis by liquid chromatography-tandem mass spectrometry. Gene ontology (GO) analysis, pathway analysis, SMART, and SABLE were used to study the possible functions of these peptide according to their precursor proteins' function. The effects of peptides derived from VCAM-1 (PDFV) on endometrial stromal cell (ESC) migration and invasion were examined with wound healing assays and Transwell assays and the expression of E-cadherin was detected by western blotting.
A total of 491 peptides were identified with abundant differences between the two groups of samples (p < 0.05, and absolute fold change ≥ 2). SMART and SABLE database showed that 42 of the 491 peptides were located in the conserved structural domains of their protein precursors and contained secondary structure and, among them, 2 peptides' precursor proteins were associated with the cell proliferation. Additionally, 5 peptides' precursor proteins were associated with endometriosis. Our study confirmed that PDFV promoted ESC migration and invasion and reduced E-cadherin expression (p < 0.05).
PDFV and its precursor protein VCAM-1 may be involved in the process of OvE formation by reducing the expression of E-cadherin. The peptidomics analysis provides new insight for future studies of the mechanisms of OvE development.
背景/目的:卵巢子宫内膜异位症(OvE)是一种内衬子宫内膜组织的卵巢囊肿。在17%-44%的子宫内膜异位症女性中可发现此类囊肿。其临床表现包括盆腔疼痛、痛经、性交困难和不孕。尽管OvE的发病率逐年上升,但其确切发病机制仍不清楚。我们运用蛋白质组学的一个新兴分支——肽组学,来鉴定差异表达的肽,以确定这些肽在OvE发病机制中的可能作用。
使用10 kDa分子量截留滤器从OvE的异位和在位子宫内膜中提取肽,然后用PEAKS软件鉴定肽前体蛋白,接着用TMT标记法进行定量,并随后通过液相色谱-串联质谱分析。根据肽前体蛋白的功能,利用基因本体(GO)分析、通路分析、SMART和SABLE来研究这些肽的可能功能。采用伤口愈合试验和Transwell试验检测血管细胞黏附分子-1衍生肽(PDFV)对子宫内膜基质细胞(ESC)迁移和侵袭的影响,并通过蛋白质印迹法检测E-钙黏蛋白的表达。
共鉴定出491种在两组样品间存在显著差异的肽(p < 0.05,绝对变化倍数≥2)。SMART和SABLE数据库显示,491种肽中的42种位于其蛋白质前体的保守结构域中,并含有二级结构,其中2种肽的前体蛋白与细胞增殖相关。此外,5种肽的前体蛋白与子宫内膜异位症相关。我们的研究证实,PDFV促进了ESC的迁移和侵袭,并降低了E-钙黏蛋白的表达(p < 0.05)。
PDFV及其前体蛋白VCAM-1可能通过降低E-钙黏蛋白的表达参与OvE的形成过程。肽组学分析为未来OvE发病机制的研究提供了新的见解。