Reduce The APC, Taxol DKK-3 Gene Methylation Caski Cell Proliferation Inhibition

Objective: To study the effects of paclitaxel on the methylation level of APC and DKK-3 gene and cell proliferation in human cervical cancer Caski cells. To provide demonstration about DNA methylation in the diagnosis and treatment of cervical cancer.Methods: 1. Paclitaxel with concentration 0,1,5,10,20,40,80,160,320 μmol/L were used to treat cervical cancer Caski cells. The growth curve was obtained through MTT method.2. After Caski cells were treated by 20 μmol/L paclitaxel and the grow inhibition was observed in different time points 24 h, 36 h, 48 h.3. After Caski cells were treated by paclitaxel with different concentration, the m RNA level were detected by RT-PCR in Caski cells. After Caski cells were treated by 20 μmol/L paclitaxel for 24 h, 36 h, 48 h, the m RNA level were detected by RT-PCR in Caski cells.4. After Caski cells were treated by paclitaxel with different concentration, the methylation level of APC and DKK-3 were detected by Methylation-Specific-PCR(MSP). After Caski cells were treated by 20 μmol/L paclitaxel for 24 h, 36 h, 48 h, methylation level of APC and DKK-3 were detected by MSP in Caski cells.Results: 1. The growth inhibit ratio of Caski cells were increased according to the raising of paclitaxel concentration after treated by paclitaxel with different concentration for 24 h. There were significant difference among the groups treated by paclitaxel with different concentration(P<0.05). Paclitaxel can inhibit the growtth of Caski cells in a dose-dependent manner.2. After Caski cells were treated by 20 μmol/L paclitaxel, in time points 24 h, 36 h, 48 h, the inhibit ratio were(51.1±1.5)%,(57.2±1.6)%,(64.7±2.1)%, respectively. There were significant difference among the three time points. Paclitaxel can inhibit the growth of Caski cells in a time-dependent manner.3. The expression level of APC and DKK-3 m RNA were increased in Caski cells treated with paclitaxel more than 5 μmol/L. The higher the concentration was, the higher expression level of APC and DKK-3 m RNA. But there was no obvious change in Caski cells treated with 1 μmol/L paclitaxel(p>0.05).4. In the time points 24 h, 36 h, 48 h, there were no significant difference on the methyaltion level of APC and DKK-3 gene between untreated group and the group with 1 μmol/L paclitaxel treating( p>0.05). Upon 5 μmol/L, along with the prolonging of treatment time and increasing in dosage of paclitaxel, the promoter methylation level of APC and DKK-3 gene in Caski cells decreased significantly(p<0.05). And there were significant difference among different concentrations and time points(p<0.05).Conclution:1.Paclitaxel could inhibit the growth of Caski cells in a dose- and time-dependent manner.2.Paclitaxel inhibit the growth of Caski cells may through down regulation of promoter methylation level of APC and DKK-3 and up regulation of their m RNA.