The Effcet Of5-aza-2’-deoxycytidine On MiR-148a Expression, CpG Island Methylation Status,and Proliferation Of Pancreatic Cancer BxPC-3Cells

Objective: To explore the effcet of5-Aza-2’-deoxycytidine on miR-148aexpression, CpG island methylation status, and proliferation of pancreatic cancerBxPC-3cells.Methods: Pancreatic cancer BxPC-3cells were cultured in DMEM and dividedinto two groups. The experimental group was treated with5μmol/L5-Aza-CdR, andthe control group was treated with the same volume of PBS. Cells were collectedafter72h. The expression of miR-148a was detected by real-time quantitativepolymerase chain reaction (RT-qPCR). The CpG island methylation status ofmiR-148a gene promotor region was measured by methylation-specific PCR (MSP).Cell proliferation was examined by MTT assay.Results:(1) The relative expression of miR-148a in experimantal group was2.93±0.11fold than that in control group. The expression level of miR-148a inexperimental group was significantly up-regulated compared with control group(P<0.01).(2) The miR-148a gene promotor CpG island was negative methylation inexperimental group, while that was positive methylation in control group.(3) The cellproliferation activities of experimental group were significantly lower than those ofcontrol group at24h,48h,72h,96h and120h after treatment (P<0.05). The growthinhibition ratio were2.36%,5.55%,15.63%,16.63%and18.90%respectively.Conclusions: Demethylation agent5-Aza-CdR can lead to demethylation ofmiR-148a gene promotor CpG island, thus up-regulate the expression level ofmiR-148a and inhibite cell proliferation in pancreatic cancer BxPC-3cell. Theexpression level of miR-148a may regulated by DNA methylation in pancreaticcancer, and act as tumor suppressor gene. miR-148a is expecetd to be a potentialeffective therapeutic target in pancreatic cancer.