May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Changes in the proliferation behaviour of a human uveal melanoma cell line from different steps in the metastatic cascade
Author Affiliations & Notes
  • J.J. Cools–Lartigue
    Microbiology/Immunology, Henry C Whitelson Ocul Path Lab, Montreal, PQ, Canada
  • J.–C.A. Marshall
    Microbiology/Immunology, Henry C Whitelson Ocul Path Lab, Montreal, PQ, Canada
  • P.L. Blanco
    Microbiology/Immunology, Henry C Whitelson Ocul Path Lab, Montreal, PQ, Canada
  • V.S. Saraiva
    Microbiology/Immunology, Henry C Whitelson Ocul Path Lab, Montreal, PQ, Canada
  • M.N. Burnier Jr
    Microbiology/Immunology, Henry C Whitelson Ocul Path Lab, Montreal, PQ, Canada
  • E. Antecka
    Microbiology/Immunology, Henry C Whitelson Ocul Path Lab, Montreal, PQ, Canada
  • Footnotes
    Commercial Relationships  J.J. Cools–Lartigue, None; J.A. Marshall, None; P.L. Blanco, None; V.S. Saraiva, None; M.N. Burnier Jr, None; E. Antecka, None.
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 1196. doi:
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      J.J. Cools–Lartigue, J.–C.A. Marshall, P.L. Blanco, V.S. Saraiva, M.N. Burnier Jr, E. Antecka; Changes in the proliferation behaviour of a human uveal melanoma cell line from different steps in the metastatic cascade . Invest. Ophthalmol. Vis. Sci. 2004;45(13):1196.

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

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Abstract

Abstract: : Purpose: Approximately forty five percent of uveal melanoma patients will die within ten years of local treatment due to metastatic disease. Cellular changes between each step of the metastatic cascade are not well characterized. The aim of this study is to describe the changes in the proliferation rate of a human uveal melanoma cell line (92.1) in successive steps of the metastatic cascade of a rabbit model. Methods: Two albino New Zealand rabbits were immuno suppressed with 15mg/kg/day intra–muscular cyclosporin A injections for three days before 1x106 92.1 cells were injected into the suprachoroidal space. Cyclosporin A doses were maintained for four weeks, and then decreased to 10mg/kg/day until nine weeks after cell implantation when the animals were sacrificed. Cells were then immediately harvested from the intraocular tumour, and the primary foci of lung metastasis. Malignant cells in the peripheral blood were isolated using the Ficoll–plaque method. Cells were re–cultured in RPMI 1640 medium supplemented with 5% fetal bovine serum, and were utilized for the proliferation assay within two passages. The sulforhodamine–B proliferation assay was used as per the National Cancer Institute method. Briefly, all cells were seeded at a concentration of five thousand cells per well in two ninety–six well plates. Plates were incubated for 72 hours before total cellular protein content per well was assessed. The averaged results per cell line were recorded. The Student’s t– test was used to compare results from control 92.1 cells and 92.1 cells cultured from each location. A p value of less than 0.05 was considered statistically significant. Results: In the first rabbit, no statistically significant changes were seen in the proliferation rates of cells re–cultured from the intraocular tumor and circulating malignant cells when compared to the control. A 51.2% increase in proliferation was seen in cells re–cultured from the lung metastasis (p<0.01). In cells re–cultured from the second rabbit, a 19.6% decrease in the proliferation rate was seen in the intraocular tumor compared to control (p<0.01). In addition, an increase in proliferation was seen in cells re–cultured from peripheral blood (38.5%, p<0.01) and lung metastasis (11.8%, p<0.01). Conclusions: This human uveal melanoma cell line displayed increased proliferation in the metastatic site. This finding indicates that cytoskeletal changes, local factors and/or a combination of both play a major role in the ability of cells to disseminate and grow in the metastatic site.

Keywords: melanoma • proliferation 
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