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.