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S. Landreville, V. Boily, S. Champagne, Québec Uveal Melanoma Group, A. Rousseau, C. Salesse; Repression of Genes Associated With Apoptosis, Development and Pigmentation in Uveal Melanoma. Invest. Ophthalmol. Vis. Sci. 2007;48(13):4775.
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There is increasing evidence that cancer can arise from a cancer stem cell, a tumor-initiating cell that has properties similar to those of stem cells. We demonstrated previously that uveal melanoma cells contain a small subpopulation of cells that exhibit a cancer stem cell-like phenotype. Subtractive library was thus prepared to identify genes which are down-regulated in the primary tumor when compared to the normal tissue.
Suppressive Subtractive Hybridization (SSH) was used to isolate genes that are repressed in uncultured primary tumors of uveal melanoma compared to normal uveal melanocytes. A library was thus constructed from the resulting subtracted cDNAs. Genes associated with apoptosis, development and pigmentation were then confirmed to be down- regulated in uncultured primary tumors using semi-quantitative RT-PCR. We also studied the methylation status of some of these genes by performing methylation-specific polymerase chain reaction (MSP-PCR).
The SSH library allowed to identify 112 genes repressed in primary uveal melanoma tumors. Semi-quantitative RT-PCR analyses confirmed the repression of selected genes implicated in apoptosis, development and pigmentation. MSP-PCR demonstrated hypermethylation of the promoter of some of these genes.
The analysis of the subtracted library revealed genes that are repressed in primary tumor of uveal melanoma. Some of these genes are associated with apoptosis, development and pigmentation and thus provided additional evidence that uveal melanoma can arise from a cancer stem cell.
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