Dioscin induces demethylation of DAPK-1 and RASSF-alpha genes via the antioxidant capacity, resulting in apoptosis of bladder cancer T24 cells

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2017-02-17

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DNA methylation at CpG rich regions often occurs at tumor suppressor gene promoters, resulting in reduced gene expression and final carcinogenesis. Hypermethylation of tumor suppressor genes, including DAPK-1 and RASSF-1α genes, have been found in patients with bladder carcinoma (BC) in some western countries. Reactive oxygen species (ROS) was reported to play a causative role in gene hypermethylation. In this study, we detected the methylation status and expression of DAPK1 and RASSF-1α genes in tissue samples from Chinese BC pa- tients, using methylation-specific PCR, reverse transcription PCR and western blotting. Further, we examined the ability of dioscin, a natural antioxidant, to regulate methylation status and expression of DAPK-1 and RASSF-1α genes in BC cell lines. In our results, DAPK-1 and RASSF-1α genes showed higher methylation level and lower express level in BC tissues than matched normal tissues. Treatment with dioscin decreased viability of BC 5637 and T24 cells, but not non-cancer bladder epithelial cell, SV-HUC-1. Dioscin triggered demethylation of DAPK1 and RASSF-1α genes in T24 cells and increased the gene and protein expression in 5637 and T24 cells. Both dioscin and substituted antioxidants (N-acetyl cysteine and Vitamin E) decreased intracellular ROS, but the effect of dioscin was abolished by adding H2O2. Similar to dioscin, the substituted antioxidants also induced the gene demethylation and T24 cell apoptosis. Co-treatment with dioscin and H2O2 had no such effects. Collectively, di- of DAPK-1 and RASSF-1α genes via the antioxidant capacity, resulting in apoptosis of bladder cancer T24 cells or inhibitory cell viability.

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dioscin, methylation, DAPK-1, RASSF-1α, antioxidant capacity, bladder cancer

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