Up-regulation Of MiR-148a Expression In Pancreatic Carcinoma AsPC-1Cells And Its Effects On DNMT1Gene Expression,Cell Proliferation And Apoptosis

Objective To investigate the effects of the upregulated expression ofmicroRNA-148a(miR-148a)on the expression of DNMT1gene,proliferation andapoptosis of Pancreatic cancer cells,Future application of RNAi in the gene therapyof Pancreatic cancer might be expected．Materials and Methods miR-148a was transfected to Pancreatic cancerAsPC-1cells by the plasmids(pcDNA3.1/pri-miR-148a)which were constructed invitro and then cells expressing the plasmids stably were yielded．There were threegroups:negative control group(transfected with pcDNA3.1),and blank controlgroup(untransfected),experimental group(transfected with pcDNA3.1/pri-miR-148a).The DNMT1mRNA and DNMT1ptotein were detected by real-time quantitativefluorescent PCR(qRT-PCR)and Western blotting, respectively；The growth status ofcells was detected by MTT assay；Flow cytometry(FCM)was used to observe the cellapoptosis and cell cycle．Results After the plasmid expressing microRNA transfected in pancreaticcancer AsPC-1cells with stable expression of miR-148a, Compared.with the blankcontrol group and negative control group, the relative DNMT1mRNA expression ofexperimental groups was significantly lower than in negative control group or thanblank control group(0.3336±0.0839VS1.0016±0.0283VS1.0000±0.0000,allP<0.05); The expression of DNMT1protein in pancreatic cancer AsPC-1cells wassignificantly reduced, P<0.05; On the fifth day, cell proliferation was repressed to24.7%; Cells in G0/G1phase of the cell ratio is high up to72.5%, higher than thosein blank control group and negative control group in AsPC-1cells in G0/G1phaseratio (respectively63.24%and63.42%)(P<0.05); cell apoptosis in experimentalgroup increased significantly, the apoptosis rate of the experimental group was4.32times more than the control group(P<0.05).Conclusions By transfecting the plasmid(pcDNA3.1/pri-miR-148a)expressing miR-148a to Pancreatic cancer AsPC-1cells,. the expressions of DNMT1mRNA and DNMT1protein Can be down-regulated.and it can inhibit pancreatic cancer cell proliferation and promote apoptosis.The miR-148a may be a target genefor pancreatic cancer treatment.