Effects of 5-Aza-dcR and TSA on methylation of transcription regulation of Syk gene in colorectal cancer cell line

Abstract

Objective To explore the effects of5-Aza-2-deoxyeytidine (5-Aza-dcR) and Tricho-statin A (TSA) on Spleen tyrosine kinase (Syk) gene expression, cell proliferation and infiltration in colo-rectal cancer cell line HCT-15. Methods The HCT-15 cells were treated with 2.5 μmol/L 5-Aza-dcR, 1 μmol/L TSA or together. Syk gene methylation status and expression level were detected by MSP, RT-PCR and/or Western blot. HCT-15 cell proliferation and in-vitro cell infiltration ability were detected by 3H in-corporation assay and in-vitro cell infiltration assay. Results (1) Compared with the control group, Syk gene was demethylated and restored expression in the presence of 5-Aza-dcR and/or TSA and TSA en-hanced the effect of 5-Aza-dcR-induced demethylation; (2) Compared with Syk (-) HCT-15 cell group in the absence of 5-Aza-dcR and/or TSA,the cell proliferation ability in groups of 5-Aza-deR,TSA, com-bined treatment,or Syk stable expression was reduced by 30% ,25% ,45% and 54% respectively. Mean-while,cell infiltration ability was reduced by 22% ,27% ,43% and 64% respectively, Conclusion Ad-ministration of 5-Aza-dcR and/or TSA can restore the re-expression of methylated Syk gene, and inhibit the proliferation, invasion and metastasis of HCT-15 cells. Key words: 5-Aza-2-deoxycytidine ; Trichostatin A; Spleen; Tyrosine kinase ; Methyla-tion