Expression Of The HIC-1 In Human Cervical Carcinoma Cell Line Hela And Its Regulation By Methylation

Objective To detect the expression of HPV and HIC-1 gene and their relations and study proliferative changes in HeLa cells with the effects of 5-Aza-2'-deoxycytidine (5-Aza-CdR).Then to investigate the promoter methylation status of HIC-1 and the potential mechanism of its antitumorigenesis in cervical cancer.Methods Human cervical cancer cell line Hela was treated with 5-Aza-CdR (5, 10 and 20μmol/L ), a specific demethylation agent for 5 days. The expression of HIC-1 mRNA and HPV18E6 mRNA was observed by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). The anti- proliferative effect was measured by methabenzthiazuron (MTT) assay.The data was analyzed with SPSS11.5 statistical software.Results After 5-Aza-CdR was added into HeLa cells with different dose for 5 days, our results were spread out below:1,After 5,10and20μmol/L 5-Aza-CdR treatment, the level of HIC-1 mRNA and P53 mRNA expression were increased, and they were significantly correlated with the concentration of 5-Aza-CdR. The expression of HPV18E6 mRNA was remaining unchanged in all of the cases.2,Hela cells treated with 5-Aza-CdR displayed a slow growth rate in comparison with that of the control cells, The cell vitalities of treated group were 91.22%±2.7%,65.08%±4.06%,49.68%±1.49%,38.80 %±3.00% individually compared with the control group and the inhibiting effect of 5-Aza-CdR was dose-dependent.3,5-Aza-CdR could reduce the density of Hela cells ,and the convergence of team was about 70 percent of the control group, the morphologic changes in cells was no significant.Conclusion1,Aberrant methylation of the HIC-1gene seems to be an important mechanism underlying its down-regulation of expression.2,Aberrant methylation of the HIC-1 seems to be one of the role in the proliferation of Hela cells .3,5-Aza-CdR can inhibit the proliferation of Hela cells with dose-dependent relationgship of its effect.