Methylation of GSTP1 promoter region in hepatocellular injury induced by isoniazid / 国际药学研究杂志

ABSTRACT

Objective To investigate the relationship between methylation of CpG island in glutathione S-transferase P1(GSTP1) promoter region and injury induced by isoniazid in HL-7702 cells.Methods HL-7702 cells were divided into the control group and three isoniazid groups(200,400,800 mg/L).Colorimetric method was used to detect the activity level of lactate dehydrogenase in the medium of HL-7702 cells;the mRNA expression of GSTP1,DNA methyltransferases 1(DNMT1),DNMT3a and DNMT3b were de?tected by real-time fluorescence quantitative PCR;the protein expression levels of DNMT1,DNMT3a and DNMT3b were detected by enzyme-linked immunosorbent assay;the methylation of the CpG island in the GSTP1 promoter region was determined by the bisulfite sequencing PCR.Results The activity level of lactate dehydrogenase in supernatants of the HL-7702 cells in isoniazid group(400, 800 mg/L)was higher than that in the control group(P<0.01).Compared with the control group,the mRNA expression of DNMT 1、3a、3b and GSTP1 were elevated in 400 and 800 mg/L isoniazid groups(P<0.05,P<0.01).The proteins expression of DNMT1 and 3a in the 400 and 800 mg/L isoniazid groups were higher than that in the control group(P<0.05,P<0.01),and the protein expression of DNMT 3b in the 800 mg/L isoniazid groups were higher than that in the control group(P<0.01).The methylation level of CpG is?land in GSTP1 promoter region of three isoniazid groups were decreased.Conclusion The CpG island in the promoter of GSTP1 has hypomethylation in hepatocyte cells damaged by isoniazid.