Effect of Aβ25-35 on Bcl-2, Bax gene promoter DNA methylation in SH-SY5Y cell
10.3969/j.issn.1001-1978.2016.08.025
- VernacularTitle:Aβ25－35对SH－SY5Y细胞Bcl－2、Bax基因启动子区DNA甲基化的影响
- Author:
Min GUO
;
Xianwu ZHANG
;
Tianyu MA
;
Hong ZHANG
;
Miao SU
;
Yan WU
;
Gang LI
- Publication Type:Journal Article
- Keywords:
SH-SY5 Y cells;
amyloid-β;
apoptosis;
DNA methylation;
DNMTs;
Bcl-2;
Bax
- From:
Chinese Pharmacological Bulletin
2016;32(8):1158-1164
- CountryChina
- Language:Chinese
-
Abstract:
Aim To investigate whether the effect of Aβ25-35 on Bcl-2 and Bax gene transcription through DNA methylation in SH-SY5Y cell.Methods Differ-ent concentrations of Aβ25-35 (0, 25, 50 μmol? L-1 ) were treated with SH-SY5Y cells for 48 h or 72 h in vitro.The optimal concentration and time of Aβ25-35 in-duced SH-SY5 Y apoptosis were determined by MTT method.Protein expression levels of Bcl-2 and Bax of Aβ25-35-treated groups were determined by Western blot.Real time PCR was used to detect the mRNA lev-els of DNA methyltransferase including DNMT 1 , DN-MT3a, DMT3b, MeCP2. Methylation specific PCR ( MSP) was used to analyze the effect of Aβ25-35 media-ted Bcl-2 and Bax gene promoter methylation .Results 25 μmol? L-1 Aβ25-35 was exposed to SH-SY5Y cells for 72 h, MTT assay showed that cell viability was (68.49 ±9.83 )%, which was significantly reduced compared with the control group ( P <0.05 ) , indica-ting AD cell apoptosis model was successfully estab-lished.Bcl-2 expression of Aβ25-35-treated group was significantly reduced compared with the control group , on the contrary , the expression of Bax was significantly increased .Real-time PCR results showed that com-pared with the control group , DNMT1, DNMT3a, DMT3b, MeCP2 mRNA levels of the Aβ25-35-treated groups had no significant difference ( P>0.05 ); MSP results showed that Bcl-2 and Bax unmethylated ampli-fication was positive , methylated amplification was neg-ative in control group , Bcl-2 and Bax unmethylated amplification was positive and methylated amplification was negative in Aβ25-35-treated group.Conclusion DNA methylation of Bcl-2 and Bax gene promoter are not affected during Aβ25-35 induced SH-SY5Y cell ap-optosis .
