May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
VEGF-D, Not VEGF-C, Expression is Regurated by Cell- Matrix Contact in Cultured Human RPE Cells
Author Affiliations & Notes
  • Y. Ikeda
    Dept Ophthalmology, Graduate Sch Med Sci Kyushi Univ, Fukuoka, Japan
  • Y. Yonemitsu
    Dept Pathology, Graduate Sch Med Sci Kyushi Univ, Fukuoka, Japan
  • M. Miyazaki
    Dept Pathology, Graduate Sch Med Sci Kyushi Univ, Fukuoka, Japan
  • R. Kohno
    Dept Pathology, Graduate Sch Med Sci Kyushi Univ, Fukuoka, Japan
  • T. Ishibashi
    Dept Pathology, Graduate Sch Med Sci Kyushi Univ, Fukuoka, Japan
  • K. Sueishi
    Dept Pathology, Graduate Sch Med Sci Kyushi Univ, Fukuoka, Japan
  • Footnotes
    Commercial Relationships  Y. Ikeda, None; Y. Yonemitsu, None; M. Miyazaki, None; R. Kohno, None; T. Ishibashi, None; K. Sueishi, None.
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 2276. doi:
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      Y. Ikeda, Y. Yonemitsu, M. Miyazaki, R. Kohno, T. Ishibashi, K. Sueishi; VEGF-D, Not VEGF-C, Expression is Regurated by Cell- Matrix Contact in Cultured Human RPE Cells . Invest. Ophthalmol. Vis. Sci. 2003;44(13):2276.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose: Vascular endothelial growth factor (VEGF) family plays an essential role in vascular development, angiogenesis and lymphangiogenesis in vivo. Among these, both VEGF-C and VEGF-D are known to stimulate angiogenesis and lymhangiogenesis via the VEGF receptor-2 (VEGFR-2/KDR) and VEGFR-3 (Flt-4). Regulational mechanism of expression, however,.has not been fully defined. We here assessed that the effect of cell-cell or cell-matrix interaction on the transcriptional regulation of VEGF-C and -D. Methods: Human retinal pigment epithelium cell line (ARPE-19) was used in this study. In several conditions, ARPE-19 was grown in a 1:1 mixture of Dulbecco's modified Eagle's medium (DMEM) and Ham's F12 medium with 3 mM L-glutamine and 10% fetal bovine serum (FBS). Total cellular RNA was extracted from cells using the Isogen-chloroform mixtured solution, and the expression of VEGF-C or VEGF-D mRNA was evaluated by semi-quantitative RT-PCR. Results: VEGF-D mRNA expression was strongly up-regulated in the presence of 2.2mM EGTA (deprivation of Ca2+ from the culture medium). When culture plate was coated with poly-HEME, an inhibitor for cell adhesion, VEGF-D expression was similarly upregurated, compared to that in cells without coating. The treatment with Ca2+ channel blockers (20uM nifedipine or 10uM diltiazem), however, did not affect the VEGF-D mRNA expression, suggesting that VEGF-D expression may be regulated by extracellular Ca2+, but not, Ca2+ influx . Meanwhile, no significant change of the VEGF-C expression was found under these culture conditions. Conclusions: These data demonstrated that VEGF-D, but not VEGF-C, mRNA was strongly induced by depletion of extracellular Ca2+ from the culture medium and did not be affected by calcium influx. These findings suggest that VEGF-D expression can be regulated by cell-matrix and/or cell-cell contact, and may imply a potential angiogenic/lymphangiogenic potential of proliferating RPE cells when the.RPE layer is broken down.

Keywords: retinal pigment epithelium • gene/expression • extracellular matrix 
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