ABSTRACT
ObjectiveTo investigate the effects of 5-Aza-CdR on the transcriptional regulation through methylation of the DNA promoter protocadherin 8(PCDHg) gene in pancreatic cancer cell line Capan-2.The Capan-2 retardation in growth rate and apoptosis were assessed in when administered 5-Aza-CdR and the chemotherapy agent,gemcitabine.MethodsMTT and flow cytometry were used to analyze the cell growth inhibition and apoptosis when treated with 5-Aza-CdR or in combination with gemcitabine.Methylation-specific PCR,RT-PCR and western blot were performed to detect methylation state,mRNA and protein respectively of PCDH8 gene in 5-Aza-CdR-treated Capan-2cells.Results Capan-2 cells treated with 5-Aza-CdR showed a slower growth rate,and a significant growth inhibition when given both 5-Aza-CdR in combination with gemcitabine.Compared with single drug administration and control,5-Aza-CdR together with gemcitabine can induce a stronger apoptosis signal.Different concentrations 5-Aza-CdR of were able to reverse methylation,restore mRNA and protein levels of PCDH8 in Capan-2.Conclusion5 Aza-CdR may demethylate the PCDH8 gene,which would effectively remove the gene silencing caused by high methylation,and thus induce gene mRNA transcription and protein expression to inhibit cell growth and have collaborative antitumor functions with gemcitabine.