The role of APAF-1 DNA methylation in osteosarcoma cell lines

Abstract

Objective To explore the role of APAF-1 DNA methylation in osteosarcoma cell lines through the detection of the methylation status of the APAF-1 promoter and the expression of the Apaf-1 mRNA in osteosarcoma cell lines. Methods Two osteosarcoma cell lines MG63 and Hs888T were used in this study. The methylation pattern in the CpG island of APAF-1 promoter was analyzed by techniques of combined bisulfite restrict analysis (COBRA). The mRNA expression of APAF-1 gene in the two cell lines treated with different concentrations of DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (5-Aza-CdR) (0,1×10~(-7) and 3×10~(-7) mol/L) was assayed by QT-PCR. Results CpG island methylation of APAF-1 gene was found in Hs888T cell line. The expression of Apaf-1 mRNA in Hs888T cell line was in-creased after the treatment of 5-Aza-CdR. There was significant difference between Hs888T cell line treated with 3×10~(-7) mol/L of 5-aza-CdR and controls (P<0.05). Conclusion CpG island methylation of APAF-1 gene in the osteosarcoma cell line Hs888T inhibits the expression of Apaf-1, which implies that APAF-1 DNA methylation might participate in the genesis of some osteosarcomas. Key words: Osteosarcoma; APAF-1; Methylation