Abstract
Purpose: :
To obtain a comprehensive view of the transcriptional profile relative to tumorgenicity in human uveal melanoma
Methods: :
Total RNAs isolated from a normal human uveal melanoctye cell line, primary uveal melanoma cell lines Mel290 and Mel270, formalin-fixed paraffin-embedded (FFPE) primary uveal melanoma and normal uveal tissue adjacent to the melanoma were used for genome-wide transcriptome analysis on Affymetrix U133 plus 2.0 microarrays. Real-time quantitative RT-PCR and immunohistochemistry was performed to validate the differentially expressed genes identified by the microarray assay.
Results: :
Hierarchical cluster analysis of the expressed genes showed that 2646 probe sets in melanocyte versus melanoma cells and 2365 probe sets in normal uveal tissue versus uveal melanoma were differentially expressed. Cross-comparison analysis showed that the expressions of 299 probe sets targeted on 79 genes have no difference between cultured cells and FFPE tissue. We found 20 genes that were consistently differentially expressed between cancerous and non-cancerous cells/tissue. Furthermore, abnormal expression of some relevant genes such as MMP2 and TIMP3 were validated by real-time quantitative RT-PCR and immunohistochemistry. Overexpression of MMP2 and TIMP3 was found in 100% of uveal melanoma specimens.
Conclusions: :
Comprenhesive gene expression analysis of uveal melanoma cells/tissue and normal uveal melancytic cells/tissue revealed different gene profiles, which could be used to discriminate between uveal melanoma and non-cancerous cells/tissues. These findings may be used to facilitate the identification of molecular markers for detecting metastatic uveal melanoma and novel potential therapeutic targets for primary uveal melanoma.
Keywords: tumors • uvea • gene/expression