J Lawry1, Z Currie, M O Smith, I G Rennie. 1. Institute for Cancer Studies, University Medical School, Sheffield, UK. j.lawry@sheffield.ac.uk
Abstract
PURPOSE: Uveal melanoma continues to present problems when attempting to predict disease progression. This study attempts to identify markers indicative of the biological characteristics of cells isolated from samples of uveal melanoma, including adhesion (ICAM-1), immune reactivity (MHC Class I and II), cell cycle control (c-erbB-2, c-myc) and apoptosis control (bcl-2, p53) using dual parameter (DNA/MoAb) flow cytometry. METHODS: Sixty-three fresh tissue samples from choroidal melanomas were taken at enucleation. Samples were assayed for DNA content and cell cycle, the above antibodies together with positive (PHM-5) and negative (2 degrees FITC Ab) controls. The clinical parameters sex, age, tumour location, cell type, tumour volume and presence of metastases were compared with the results and analysed with the non-parametric Mann-Whitney U-t-test. RESULTS: ICAM-1 expression proved to be the most clinically relevant, being present on a higher proportion of cells in tumours > 2000 mm3 (median 38, n = 19) compared with the smaller tumours < 2000 mm3 (median 17, n = 26) (p = 0.0015). Metastatic disease was present in 11 patients and did not correlate with any of the surface markers. C-myc, c-erbB-2 and MHC Class II expression were associated with cell type, all showing greater expression in spindle cell tumours than mixed/epithelial types. CONCLUSION: These results show flow cytometry as a quick, easy method to provide a 'phenotypic profile' for these tumours, and identifies cell cycle control and adhesion molecule expression as important areas for further investigation. c-erbB-2 and bcl-2 positivity was typically seen on over 60% cells in each sample, indicating two potential targets for therapeutic intervention.
PURPOSE:Uveal melanoma continues to present problems when attempting to predict disease progression. This study attempts to identify markers indicative of the biological characteristics of cells isolated from samples of uveal melanoma, including adhesion (ICAM-1), immune reactivity (MHC Class I and II), cell cycle control (c-erbB-2, c-myc) and apoptosis control (bcl-2, p53) using dual parameter (DNA/MoAb) flow cytometry. METHODS: Sixty-three fresh tissue samples from choroidal melanomas were taken at enucleation. Samples were assayed for DNA content and cell cycle, the above antibodies together with positive (PHM-5) and negative (2 degrees FITC Ab) controls. The clinical parameters sex, age, tumour location, cell type, tumour volume and presence of metastases were compared with the results and analysed with the non-parametric Mann-Whitney U-t-test. RESULTS:ICAM-1 expression proved to be the most clinically relevant, being present on a higher proportion of cells in tumours > 2000 mm3 (median 38, n = 19) compared with the smaller tumours < 2000 mm3 (median 17, n = 26) (p = 0.0015). Metastatic disease was present in 11 patients and did not correlate with any of the surface markers. C-myc, c-erbB-2 and MHC Class II expression were associated with cell type, all showing greater expression in spindle cell tumours than mixed/epithelial types. CONCLUSION: These results show flow cytometry as a quick, easy method to provide a 'phenotypic profile' for these tumours, and identifies cell cycle control and adhesion molecule expression as important areas for further investigation. c-erbB-2 and bcl-2 positivity was typically seen on over 60% cells in each sample, indicating two potential targets for therapeutic intervention.