DOI: 10.36347/sajb.2016.v04i11.011

Pages: 1038-1044

The objective is to discuss the treatment effects of low doses of 5 – Azacytidine to KYSE450 esophageal cancer cell line. The cells of KYSE450 esophageal cancer cell line were used different drug concentrations of 5-Azacytidine（5-Aza）, and used Western blot hybridization technique(Western blotting) detected ofγH2AX protein expression of cells, selected drug concentration of relative to the maximum and not produce cytotoxic for subsequent experiments. Methylation-specific PCR (Methylation Specific Polymerase Chain Reaction, MSP) detected methylation status changed circumstances of DNMT1 and DNMT3b gene promoter at different time points in pretherapy and post-treatment. 3. Western blotting analysis the protein expression changed of DNMT1 and DNMT3b genes at different time points in pretherapy and post-treatment. Real-time quantitative PCR (Quantitative Real time Polymerase Chain Reaction, qRT-PCR) technique detected mRNA expression of DNMT1 and DNMT3b genes at different time points in pretherapy and post-treatment. 5. Flow cytometry is used to analyze the cell cycle change of each cell. 1. Application of 5-Aza treatment KYSE450 esophageal cancer cells after 72h, measured 1000nM/L drug concentrations relative to the maximum dose of the drug does not produce toxicity. The cells of KYSE450 esophageal cancer cell line were treated with low doses of 5-Aza, DNMT1, DNMT3b gene methylation was decreases at first and increases late with time rendering. 3. Demethylation can increase silencing the expression of DNMT1, DNMT3b genes, so that is decreased expression of the mRNA and protein, there were the same trend with decreases at first and increases late, and can lead to cell cycle arrest, inhibition of cell proliferation. Low doses of 5-Aza does not produce toxic effects on cells, and can be reversed DNMT1, DNMT3b methylation status for clinical epigenetic therapy provides long-term basis. The expression of DNMT1 and DNMT3b gene decrease with mRNA and protein and cell cycle