Mechanism of PMP24 gene silencing in a PC3 prostate cancer cell linePublic
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Numerous genetic and epigenetic changes occur in the cell during prostate carcinogenesis. Recently, the importance of epigenetic events in carcinogenesis has been emphasized. Changes in DNA methylation pattern is one of these events. Tumor cells commonly exhibit overall genome hypomethylation and local hypermethylation of many genes. PMP24 encodes a 24 kDa Peroxisomal Membrane Protein that was recently determined to be hypermethylated in two prostate cancer cell lines and to contribute to prostate cancer progression. This project studied the methylation pattern and expression of the PMP24 gene in a PC3 cell line. The effect of different concentrations of demethylating agent 5-AZA-2'-deoxycytidine (5AZAdC) and different times of treatment on the PMP24 methylation pattern and expression were studied. This study establishes a correlation between PMP24 gene CpG island hypermethylation and gene silencing in the PC3 cell line. The gene is re-expressed by 5AZAdC treatment but not by the histone deacetylase inhibitor TSA, which indicates that PMP24 gene expression is silenced solely by DNA methylation, not by histone deacetylation. The AP2 binding site within promoter CpG island of PMP24 gene was discovered to be demethylated first after the treatment with 5AZAdC, which suggests that demethylation is specific, targeting the crucial sites for transcription first.
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