July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Protein Kinase C- delta Regulates Cellular Attachment, Survival, and Metastases-Associated Genes in Uveal Melanoma
Author Affiliations & Notes
  • Ansley K Ulmer
    Biology, Furman University, Greenville, South Carolina, United States
    Ophthalmology/ Hamilton Eye Institute, University of Tennessee Health Science Center, Memphis, Tennessee, United States
  • Bradley Gao
    Ophthalmology/ Hamilton Eye Institute, University of Tennessee Health Science Center, Memphis, Tennessee, United States
  • Zachary K. Goldsmith
    Ophthalmology/ Hamilton Eye Institute, University of Tennessee Health Science Center, Memphis, Tennessee, United States
  • Anna M Walsh
    Ophthalmology/ Hamilton Eye Institute, University of Tennessee Health Science Center, Memphis, Tennessee, United States
  • Sarah E Langsdon
    Ophthalmology/ Hamilton Eye Institute, University of Tennessee Health Science Center, Memphis, Tennessee, United States
  • Kelley Yuan
    Ophthalmology/ Hamilton Eye Institute, University of Tennessee Health Science Center, Memphis, Tennessee, United States
  • Matthew W Wilson
    Ophthalmology/ Hamilton Eye Institute, University of Tennessee Health Science Center, Memphis, Tennessee, United States
    Surgery, St. Jude Children's Research Hospital, Memphis, Tennessee, United States
  • Vanessa Marie Morales-Tirado
    Ophthalmology/ Hamilton Eye Institute, University of Tennessee Health Science Center, Memphis, Tennessee, United States
    Microbiology, Immunology, and Biochemistry, University of Tennessee Health Science Center, Memphis, Tennessee, United States
  • Footnotes
    Commercial Relationships   Ansley Ulmer, None; Bradley Gao, None; Zachary Goldsmith, None; Anna Walsh, None; Sarah Langsdon, None; Kelley Yuan, None; Matthew Wilson, None; Vanessa Morales-Tirado, None
  • Footnotes
    Support  Research to Prevent Blindness, West Cancer Center, 930 Friends, The Neuroscience Institute at UTHSC, Gerwin Fellowship, Furman University Neuroscience Fellowship
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 3168. doi:
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    • Get Citation

      Ansley K Ulmer, Bradley Gao, Zachary K. Goldsmith, Anna M Walsh, Sarah E Langsdon, Kelley Yuan, Matthew W Wilson, Vanessa Marie Morales-Tirado; Protein Kinase C- delta Regulates Cellular Attachment, Survival, and Metastases-Associated Genes in Uveal Melanoma. Invest. Ophthalmol. Vis. Sci. 2018;59(9):3168.

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

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Abstract

Purpose : Uveal melanoma (UM) is the most common primary intraocular malignancy in adults. It comprises about 5% of total melanomas and it is genetically and molecularly distinct than cutaneous melanoma. Somatic mutations in the GNAQ and GNA11 genes have been found in over 80% of UM signaling through PLC and ERK pathways. We have previously shown an anti-apoptotic role of Protein Kinase C-delta (PKC-d), a novel isoform of the PKC family, which is activated by diacylglycerol and phospholipids, in UM. We also demonstrated increased expression in UM eyes compared to healthy controls. In this study we focus on the morphological changes, AKT- and NFkB- cell survival mechanisms, and the modulation of genes highly associated with metastatic progress in UM after PKCD gene silencing.

Methods : We utilized 3 different UM cell lines: Mel270, OMM2.5, and Mel290. Knockdown of PKCD expression was performed by siRNA and confirmed by qPCR and Western blot analyses. Morphological changes were evaluated by microscopy imaging among different cell culture conditions, including untreated, scramble siRNA, PKCD siRNA, and the pharmacological PKC modulator Rottlerin. AKT and MDM2 signaling were evaluated by Wb and nuclear translocation of the p65 NFkB subunit by imaging flow cytometry.

Results : We observed increase in cell detachment across all UM cell lines studied after PKCD knockdown. These morphological changes were confirmed with Rottlerin. There was a reduction in cell survival mechanisms across all UM after treatment with PKCD siRNA. We found a reduction in MMP2 and CD44, which are associated with metastatic progress in cancer.

Conclusions : Our work demonstrates PKCD knockdown could reduce tumor migration as it increases cell detachment, and reduced the expression of CD44 and MMP2. In addition, we observed a reduction in cell survival in an AKT-dependent manner. Collectively, our work suggests PKC-d could be used as a potential targeted therapy for UM.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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