April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Hemangiogenic profile of uveal versus conjunctival melanoma cell lines
Author Affiliations & Notes
  • Ludwig M. Heindl
    Department of Ophthalmology, University of Cologne, Cologne, Germany
  • Nasrin Refaian
    Department of Ophthalmology, University of Cologne, Cologne, Germany
  • Simona Luise Schlereth
    Department of Ophthalmology, University of Cologne, Cologne, Germany
  • Deniz Hos
    Department of Ophthalmology, University of Cologne, Cologne, Germany
  • Jacobus J Bosch
    Department of Internal Medicine 5, Hematology and Oncology, University of Erlangen-Nuremberg, Erlangen, Germany
  • Claus Cursiefen
    Department of Ophthalmology, University of Cologne, Cologne, Germany
  • Konrad R Koch
    Department of Ophthalmology, University of Cologne, Cologne, Germany
  • Footnotes
    Commercial Relationships Ludwig Heindl, None; Nasrin Refaian, None; Simona Schlereth, None; Deniz Hos, None; Jacobus Bosch, None; Claus Cursiefen, None; Konrad Koch, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 5065. doi:
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      Ludwig M. Heindl, Nasrin Refaian, Simona Luise Schlereth, Deniz Hos, Jacobus J Bosch, Claus Cursiefen, Konrad R Koch; Hemangiogenic profile of uveal versus conjunctival melanoma cell lines. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5065.

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

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Abstract

Purpose: Uveal and conjunctival melanoma (UM, CM) are distinct malignant entities, which show diverging patterns of metastatic spread. Hematogenic metastases, which are dependent on tumor cell driven formation of blood vessels (hemangiogenesis) within and surrounding the tumor, often occur in UM. In contrast, spreading of CM is less frequent and primarily affects regional lymph nodes. The purpose of the present study was to analyze whether these differences of hematogenic spread, which are of high prognostic relevance, could be attributable to differential hemangiogenic potentials of UM and CM cells. This was assessed by measuring (1) VEGF-A expression in UM and CM cells and (2) proliferation rates of blood endothelial cells (BECs) incubated with UM/CM cell conditioned medium.

Methods: mRNA expression of VEGF-A by human primary UM cells OCM-1 and Mel-270 as well as CM cells CM2005.1 was measured by quantitative real-time PCR. For proliferation analysis, BECs were seeded on a 96-well-plate (4000 cells/well) and were left to attach for 24 hours. Hereafter cells were incubated with 50 % of BEC medium containing 1 % fetal calve serum and 50 % UM or CM cell conditioned medium. BrdU was added 1 hour later. Colorimetric analysis was performed after 48 hours of incubation at a measuring wavelength of 450 nm.

Results: VEGF-A mRNA was expressed in all three cell lines. Mel-270 cells expressed VEGF-A mRNA to a lower extent than OCM-1 and CM2005.1 (p<0.05). The proliferation of BECs was significantly increased by adding conditioned medium of CM2005.1, OCM-1, or Mel-270 cells (p<0.05 for all groups). CM2005.1- and OCM-1-conditioned medium had a similar impact, while a smaller proliferative increase of BECs was found under the influence of Mel-270 (p<0.05).

Conclusions: Both UM and CM cells expressed VEGF-A, and increased BEC proliferation. Higher VEGF-A levels (independent of tumor cell origin) were associated with a stronger increase of proliferation. A reduced hemangiogenic potential of CM cells was not detectable. Other additional mechanisms within the tumor microenvironment might thus account for the lower hematogenic metastatic rate in CM.

Keywords: 744 tumors • 474 conjunctiva • 589 melanoma  
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