title: Effects of HLEC on the secreted proteins of epithelial ovarian cancer cells prone to metastasize to lymph nodes.
abstract: OBJECTIVE
  To study explores the effect of HLEC on the secreted proteins of epithelial ovarian cancer (EOC) cells (SKOV3-PM4) with directional highly lymphatic metastasis.
  
  
  METHODS
  Supernatants of four groups of cultured cells, namely, SKOV3 (A), SKOV3+HLEC (B), SKOV3-PM4 (C), SKOV3-PM4+HLEC (D), were collected, and their proteins were detected by antibody arrays and iTRAQ-2D-LC-MALDI-TOF/TOF/MS. Significantly differential proteins were further analyzed via bioinformatics and validated in human serums and cell media via ELISA.
  
  
  RESULTS
  Results of antibody arrays and mass spectrometry demonstrated that GRN and VEGFA were upregulated in group C (compared with group A), whereas IGFBP7 and SPARC were downregulated in group D (compared with group C). Comprehensive bioinformatics analysis results showed that IGFBP7 and VEGFA were closely linked to each other. Further validation with serums showed statistical significance in VEGFA and IGFBP7 levels among groups of patients with ovarian cancers, benign tumors, and control groups. Two proteins were upegulated in the first group. VEGFA in the control group was downregulated. For IGFBP, upregulation in the control group and down-regulation in the first group were also observed.
  
  
  CONCLUSIONS
  The HLEC microenvironment is closely associated with directional metastasis to lymph nodes and with differential proteins including cell stromal proteins and adhesion factors. The upregulation of VEGFA and GRN and the downregulation of SPARC and IGFBP7 are closely associated with directional metastasis to lymph nodes in EOC cells.
authors.affiliation: The Cancer Hospital Affiliated to Guangxi Medical University, Nanning 530021, China;
authors.forename: Xin-Ying
authors.initials: XY
authors.lastname: Zhang
authors.affiliation: The Experiment Center of The Guangxi Medical University, Nanning 530021, China.
authors.forename: Fu-Qiang
authors.initials: FQ
authors.lastname: Yin
authors.affiliation: The Cancer Hospital Affiliated to Guangxi Medical University, Nanning 530021, China;
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: The Cancer Hospital Affiliated to Guangxi Medical University, Nanning 530021, China;
authors.forename: Ting
authors.initials: T
authors.lastname: Gao
authors.affiliation: The Cancer Hospital Affiliated to Guangxi Medical University, Nanning 530021, China;
authors.forename: He-Yun
authors.initials: HY
authors.lastname: Ruan
authors.affiliation: The Cancer Hospital Affiliated to Guangxi Medical University, Nanning 530021, China;
authors.forename: Xiao
authors.initials: X
authors.lastname: Guan
authors.affiliation: The Cancer Hospital Affiliated to Guangxi Medical University, Nanning 530021, China;
authors.forename: Ying-Xin
authors.initials: YX
authors.lastname: Lu
authors.affiliation: The Cancer Hospital Affiliated to Guangxi Medical University, Nanning 530021, China;
authors.forename: Dan-Rong
authors.initials: DR
authors.lastname: Li
keywords: Ovarian cancer; human lymphatic capillary endothelial cells; lymphatic metastasis; secreted proteins; tumor microenvironment