The extra domain A of fibronectin increases VEGF-C expression in colorectal carcinoma involving the PI3K/AKT signaling pathway

PLoS One. 2012;7(4):e35378. doi: 10.1371/journal.pone.0035378. Epub 2012 Apr 9.


The extra domain A (EDA)-containing fibronectin (EDA-FN), an alternatively spliced form of the extracellular matrix protein fibronectin, is predominantly expressed in various malignancies but not in normal tissues. In the present study, we investigated the potential pro-lymphangiogenesis effects of extra domain A (EDA)-mediated vascular endothelial growth factor-C (VEGF-C) secretion in colorectal carcinoma (CRC). We detected the expressions of EDA and VEGF-C in 52 human colorectal tumor tissues and their surrounding mucosae by immunohistochemical analysis, and further tested the correlation between the expressions of these two proteins in aforementioned CRC tissues. Both EDA and VEGF-C were abundantly expressed in the specimens of human CRC tissues. And VEGF-C was associated with increased expression of EDA in human CRC according to linear regression analysis. Besides, EDA expression was significantly correlated with lymph node metastasis, tumor differentiation and clinical stage by clinicopathological analysis of tissue microarrays containing tumor tissues of 115 CRC patients. Then, human CRC cell SW480 was transfected with lentivectors to elicit expression of shRNA against EDA (shRNA-EDA), and SW620 was transfected with a lentiviral vector to overexpress EDA (pGC-FU-EDA), respectively. We confirmed that VEGF-C was upregulated in EDA-overexpressed cells, and downregulated in shRNA-EDA cells. Moreover, a PI3K-dependent signaling pathway was found to be involved in EDA-mediated VEGF-C secretion. The in vivo result demonstrated that EDA could promote tumor growth and tumor-induced lymphangiogenesis in mouse xenograft models. Our findings provide evidence that EDA could play a role in tumor-induced lymphangiogenesis via upregulating autocrine secretion of VEGF-C in colorectal cancer, which is associated with the PI3K/Akt-dependent pathway.

MeSH terms

  • Aged
  • Animals
  • Carcinoma / metabolism*
  • Carcinoma / pathology
  • Cell Line, Tumor
  • Cells, Cultured
  • Chromones / pharmacology
  • Colorectal Neoplasms / metabolism*
  • Colorectal Neoplasms / pathology
  • Enzyme Inhibitors / pharmacology
  • Female
  • Fibronectins / genetics
  • Fibronectins / metabolism*
  • Humans
  • Immunohistochemistry
  • Lymphangiogenesis / drug effects
  • Lymphatic Metastasis
  • Male
  • Mice
  • Middle Aged
  • Morpholines / pharmacology
  • Neoplasm Staging
  • Phosphatidylinositol 3-Kinases / metabolism
  • Phosphoinositide-3 Kinase Inhibitors
  • Proto-Oncogene Proteins c-akt / antagonists & inhibitors
  • Proto-Oncogene Proteins c-akt / metabolism
  • RNA, Small Interfering / metabolism
  • Signal Transduction / drug effects
  • Vascular Endothelial Growth Factor C / biosynthesis*
  • Xenograft Model Antitumor Assays


  • Chromones
  • Enzyme Inhibitors
  • Fibronectins
  • Morpholines
  • Phosphoinositide-3 Kinase Inhibitors
  • RNA, Small Interfering
  • VEGFC protein, human
  • Vascular Endothelial Growth Factor C
  • extra domain A fibronectin, human
  • 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
  • Proto-Oncogene Proteins c-akt