Effect Of Demethylation On Expression Of BLU And HPV In Human Cervical Cancer Cell Line Hela

Objective To detect the expression of HPV and BLU and cell proliferation in HeLa cells exposed to 5-Aza-2'-deoxycytidine (5-Aza-CdR).Then to investigate the promoter methylation status of BLU and the potential mechanism of its antitumorigenesis in cervical cancer.Methods Human cervical cancer cell line Hela was exposed to 5-Aza-CdR(5,10 and 20μmol/L),a specific demethylation agent for 3 days.The expression ofBLU mRNA and HPV18E6 mRNA was detected 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 3 days,Our results were spread out below:1,After 5,10and20pmol/L 5-Aza-CdR treatment,the level of BLU mRNA expression was increased,and it was significantly correlated with the concentration of 5-Aza-CdR.the expression of HPV 18E6 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 0.490±0.022,0.403±0.017,0.294±0.017 individually and the inhibiting effect of 5-Aza-CdR was dose-dependent.3,5-Aza-CdR could induce morphologic changes in Hela cells. Which indicate the weakened vitality of the cells.Conclusion1,5-Aza-CdR can up-regulate the expression of BLU,it indicate that hypermethylation of BLU may contribute to its transcription inactivation.2,Demethylation induced re-expression of BLU cannot down-regulate the expression of HPV.3,Hypermethylation of BLU may play an important role in the carcinogenesis and progression of cervical cancer.