May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
PEDF-Mediated Decrease in Uveal Melanoma Hepatic Micrometastasis: Genome-Wide Analysis of Gene Expression Profiles
Author Affiliations & Notes
  • H. Yang
    Ophthalmology, Emory University Eye Center, Atlanta, Georgia
  • Y. H. Li
    Yerkes National Primate Research Center, Emory University, Atlanta, Georgia
  • N. Wagar
    Yerkes National Primate Research Center, Emory University, Atlanta, Georgia
  • H. E. Grossniklaus
    Ophthalmology, Emory University Eye Center, Atlanta, Georgia
  • Footnotes
    Commercial Relationships  H. Yang, None; Y.H. Li, None; N. Wagar, None; H.E. Grossniklaus, None.
  • Footnotes
    Support  NCI R01 CA126557, NEI P30 EY06360 and an unrestricted grant from Research to Prevent Blindness, Inc.
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 30. doi:https://doi.org/
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    • Get Citation

      H. Yang, Y. H. Li, N. Wagar, H. E. Grossniklaus; PEDF-Mediated Decrease in Uveal Melanoma Hepatic Micrometastasis: Genome-Wide Analysis of Gene Expression Profiles. Invest. Ophthalmol. Vis. Sci. 2008;49(13):30. doi: https://doi.org/.

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

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Abstract

Purpose: : Pigment epithelium-derived factor (PEDF) exhibits indirect and direct anti-melanoma mechanisms of action. We have previously demonstrated that mouse PEDF gene transfection was able to decrease hepatic micrometastasis in a mouse ocular melanoma model. The purpose of this study is to explore the molecular mechanisms of the PEDF-mediated reduction of micrometastasis.

Methods: : pLenti6.2/N-LumioTM/V5-DEST/mPEDF was transduced into the mouse melanoma B16LS9 cell line, and pLenti6.2/N-LumioTM/V5-DEST/LacZ was transduced a control. Transduced cell clones were selected by Blasticidin for 2 weeks and maintained in culture medium. The level of PEDF expression was detected by Western blot and SYBR green real-time PCR. Gene expression profiles of stably PEDF-overexpressing, vehicle control and wild-type B16LS9 cells were analyzed with Affymetrix GeneChip Mouse Genome 430 2.0 arrays representing approximately 34,000 transcripts.

Results: : A total of 3212 transcripts exhibited at least 2-fold differences in their expression levels. PEDF gene and VEGF-A gene were up-regulated respectively 32.16-fold and 2.47-fold in the PEDF-transfected cells compared to the wild type cells, which means PEDF gene expressed 13.03-fold higher than VEGF gene in the PEDF-transfected cells. Contrarily, PEDF gene and VEGF-A gene were up-regulated respectively 1.05-fold and 4.4-fold in the LacZ-transfected cells compared to the wild type cells, and the ratio of PEDF to VEGF was 0.24. S100b, Junb and TRAF family member-associated Nf-kappa B activator (Tank) were down-regulated, and hypoxia-inducible factor 1, alpha subunit inhibitor (Hif1an), metastasis suppressor 1(Mtss1) and tissue inhibitor of metalloproteinase 2 (Timp2) were up-regulated in stable PEDF overexpressing B16LS9 cells.

Conclusions: : PEDF gene overexpression results in an increase PEDF/VEGF ratio. The mechanism of inhibiting micrometastasis may be caused by the PEDF gene overexpression that inhibits NF-ΚB which leads to decreased melanoma cell proliferation with decreased JunB and HIF which regulate melanoma angiogenesis.

Keywords: gene/expression • gene microarray • melanoma 
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