April 2010
Volume 51, Issue 13
ARVO Annual Meeting Abstract  |   April 2010
Pigment Epithelium-Derived Factor RNA Interference Promotes Ocular Melanoma Growth in a Mouse Model
Author Affiliations & Notes
  • H. Yang
    Ophthalmology, Emory University Eye Center, Atlanta, Georgia
  • H. E. Grossniklaus
    Ophthalmology, Emory University Eye Center, Atlanta, Georgia
  • Footnotes
    Commercial Relationships  H. Yang, None; H.E. Grossniklaus, None.
  • Footnotes
    Support  NIH NCI R01 CA126447 and NEI R24 EY017045
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 864. doi:
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    • Get Citation

      H. Yang, H. E. Grossniklaus; Pigment Epithelium-Derived Factor RNA Interference Promotes Ocular Melanoma Growth in a Mouse Model. Invest. Ophthalmol. Vis. Sci. 2010;51(13):864.

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

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Purpose: : Our previous studies demonstrated that overexpression of pigment epithelium-derived factor (PEDF) inhibited ocular melanoma growth and metastasis. The purpose of this study is to determine if down-regulation of endogenous PEDF affects the growth of ocular melanoma, and to know the functional consequences of PEDF down-regulation during malignant progression of ocular melanoma.

Methods: : To establish a melanoma cell line which expresses the down-regulated PEDF gene, PEDF shRNA or control shRNA lentiviral particles were transduced into mouse melanoma cell line B16LS9. After puromycin selection for 2 weeks, PEDF expression was detected by RT-PCR and western blot. CyQUANT® cell proliferation assay was performed in B16LS9 with PEDF down-regulation (B16LS9-PEDFshRNA) and B16LS9 with control shRNA transduction (B16LS9-shRNA). 2.5x105/2µl of B16LS9-PEDFshRNA and B16LS9-shRNA cells were inoculated into posterior chamber in C57/BL6 mice. At the 7th day after inoculation, the eye was checked with histological methods, and the largest area of ocular tumor was measured under microscope with Image J.

Results: : PEDF expression was found in B16LS9-shRNA. The loss of PEDF expression in B16LS9-PEDFshRNA cells was confirmed using RT-PCR and western blot. Tumor cell proliferation in vitro was increased by PEDF knockout. In vivo experiments showed the area of ocular tumor with B16LS9-PEDFshRNA was significantly larger than one with B16LS9-shRNA (P<0.01).

Conclusions: : Endogenous PEDF plays an important role in the proliferation of ocular melanoma. Loss of PEDF promotes the growth in ocular melanoma.

Keywords: melanoma • proliferation • gene modifiers 

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