Effect of 5-Aza-dC on Expression and Methylation of TFPI-2 Gene in Panc-1 Pancreatic Cancer

Objective To investigate the effects of 5-aza-2，-deoxycytidine(5-Aza-dC)，a methylation inhibitor，on the expression and methylation of TFPI-2 gene in Panc-1 cell lines of pancreatic cancer. Methods Panc-1 cell line was treated with different dosages of 5-Aza-dC.TFPI-2 gene DNA，mRNA and protein were determined by MSP，RT-PCR and Western blot respectively. Results MSP detection showed that the TFPI-2 gene hypermethylation has effectively been reversed by 5-Aza-dC.Moreover，the expression levels of TFPI-2 mRNA treated with 5-Aza-dC were increased (0.211 ± 0.087，0.327 ± 0.068 and 0.609 ± 0.017，respectively).Western blot indicated that 5-Aza-dC could recover the TFPI-2 protein expression (0.429 ± 0.121，0.675 ± 0.044 and 1.132 ± 0.124 respectively).These effects within certain extent were dose and time dependent with statistical significance (P<0.05). Conclusion The hypermethylation of promoter region was a main cause for transcriptional inactivation of TFPI-2 gene in Panc-1 cell lines.5-Aza-dC might effectively reactivate the gene transcription through a demethylation role.