Construction Of Recombinant MiR-181a Vector And Its Inhibition In Pancreatic Cancer Cells

BACKGROUND:Pancreatic cancer is a lethal disease with poor survival and high mortality,and up to now no effective therapeutic approaches have been found in blame of hardly early diagnosis,early metastasis,and resistance to chemotherapy and radiation. Accurate early diagnosis and new therapeutic modalities are therefore urgently needed. MicroRNAs(miRNAs) are small noncoding RNAs form 19 to 25 nucleotides,which negatively regulate post-transcriptional gene expression.Disturbance of microRNA expression may play a role in the metabolism,proliferation,differentiation,apoptosis of cells.K-RAS is an oncogene expressed nearly 100%in pancreatic ductal adenocarcinoma.It is the target gene of some accurancy miRNAs such as let-7 in lung cancers.MicroRNA expression patterns have been made to differentiate pancreatic adenocarcinoma from normal pancreas and chronic pancreatitis.And the target gene of certain aberrantly expressed miRNAs and the role of certain miRNAs in pancreatic cancer cell has been identified.Objective:Construct the recombinant MiR-181a vector to express miR-181 a in pancreatic cancer cell lines.Transduct the recombinant vector,comfirm whether miR-181a targeting at K-RAS oncogene and its expression has effect in inhibiting pancreatic cancer cell growth.Methords:Construct the recombinant vector pcDNA4.0-EGFP-MiR-181a to express miR-181a and vector pcDNA4.0-EGFP as control.Transfect the recombinant vector in pancreatic cancer cell lines,comfirm whether miR-181a targeting at K-RAS oncogene and its expression has effect in inhibiting pancreatic cancer cell growth.Result:Expression of miR-181a has effect in inhibiting the K-RAS protein in pancreatic cancer cells.Expression of miR-181a has effect in inhibiting pancreatic cancer cell growth in vitro; Expression of miR-181a has effect in promoting pancreatic cancer cell apoptosis in vitro;Expression of miR-181a has effect in inhibiting pancreatic cancer cell migration in vitro;Conclusion:Expression of miR-181a has effects in inhibiting pancreatic cancer cell growth and inducing cell apoptosis in vitro and inhibits K-RAS expression in pancreatic cancer cell lines.