Adenose Intervenes Methylation Of RECK Gene In Human Colorectal Cancer SW480 Cells

Objective:To determine the effect of adenosine on the methylation of the reversioninducing-cysteine-rich protein with Kazal motifs (RECK) gene in human colorectal cancer SW480 cells and investigate the underlying mechanism.Methods:SW480 cells were treated with adenosine at a dose of 0 or 3.0 mmol/L for 72 h,and then the CpG islands methylation levels of the RECK gene promoter region were detected by Bisulfite sequencing polymerase chain reaction (BSP), The RECK expression at mRNA and protein levels,the methyltransferase (DNMT1 and DNMT3a) protein expression and cell apoptosis before and after adenosine processing were detected by RT-PCR, Western blotting,and flow cytometry respectively.Results:After the intervention of adenosine, the RECK gene methylation levels was significantly lower than that of the control group [(49.14 ± 2.38)% vs (74.84 ± 2.38)%, P< 0.01]. The mRNA and protein expression levels of RECK and the apoptotic rates were all significantly higher than the control group [0.3353 ±0.3502 vs 0.0800 ± 0.0036,0.6030 ± 0.0178 vs 0.0900 ± 0.0030, (27.90 ± 2.32)% vs (5.09 ±0.30)%, all P <0.01]. The protein levels of methyltransferase DNMT1 and DNMT3a were significantly lower than the control group (0.1413 ± 0.0021 vs 0.645 0 ± 0.0092,0.1300 ± 0.0046 vs 0.5267 ±0.0095, both P< 0.01).Conclusion:Adenosine inhibits methyltransferase activity to reduce the DNA methylation of RECK promoter region, then enhances the mRNA and protein levels of RECK, and thus prompts the apoptosis of SW480 cells.