The Research On DAC Accelerates Hela Apoptosis Of Cervical Carcinoma And Demethylation Of APC Gene

Objectives:To explore the effect of DAC on Hela cells apoptosis and APC gene methylation. To provide theoretical basis and experimental foundations of DAC applying to clinical treatment in cervical cancer.Methods:(1) DAC with different concentrations were used in cervical carcinoma Hela cells for 24 h and 36 h, 48 h.MTTs were performed to detect the proliferation inhibition effects in each group to screen the best concentration and time of proliferation inhibition for DAC in Hela cell;(2). Annexin V-FITC/PI double staining was used to detect DAC’s apoptosis influence on cervical carcinoma He La cell; PI single staining flow cytometry to detect DAC’s cell cycle influence on cervical carcinoma He La cells;(3). Using Methylation specific PCR(Methylation specific PCR, MSP) to detect the Methylation status of gene APC’s 5 ’end promoter region Cp G island, before and after the treatment of DAC in cervical carcinoma He La cells,;(4).RT-PCR was performed to detect APC m RNA’s expression in He La cells with or without DAC treatment.(5). Using western blot to detect the expressions of protein APC and protein β-catenin in He La’s intracellular and nuclear.Results:1. The inhibition of DAC to Hela cells hyperplasia of cervical carcinoma depend on concentration & time.2. It is proved that DAC is able to accelerate the apoptosis of Hela cells. The apoptotic rate is improved from 12.41±0.24% to 73.82±0.11%. The difference is of statistical significance(P<0.05); DAC can retard the Hela cells of cervical carcinoma at the stage of S & G2.3. After the treatment of Hela cells with DAC, the demethylation of promoter region of APC gene increases severly.4. After the treatment of Hela cells with DAC, the expression of APC m RNA increases & the difference is of statistical significance(P<0.05).5. The relative expressions of APC protein in Hela cells are 0.780 and 1.542 before and after the treatment with DAC; the relative expressions of intacellunar β-catenin protein are 0.89 and 0.354 before and after the treatment; the relative expressions of intranuclear β-catenin protein are 0.632 and 0.336 before and after the treatment. Statistical significance is proved on the difference of APC protein and intracellunar / intranuclear β-catenin protein(P<0.05).Conclusion:1. DAC is able to inhibit the hyperplasia of cervical carcinoma cells in the concentration & time dependence manner.2. DAC is able to accelerate the apoptosis of Hela cells, which leads to retardant at the stage of S & G2.3. DAC can affect the demethylation of APC gene, decrease of expression of intracellunar of APC protein, intracellunar / intranuclear β-catenin, block the signal channel of WNT, which lead to the apoptosis of cervical carcinoma cells.