Mechanism of circBFAR regulating hsa-miR-424-5p and PI3K-Akt signaling pathway genes to promote the progression of pancreatic cancer
Objective To investigate the differential expression of circBFAR in pancreatic cancer and its molecular mechanisms of action.Methods The differential expression of circBFAR in pancreatic cancer and control tissues was analyzed in the GSE69362 dataset.Differentially expressed miRNAs(DEmiRs)between pancreatic cancer and control tissues were identified using the TCGA database,and an intersection analysis was performed with circBFAR target miRNAs.Differentially expressed mRNAs(DEmRs)between pancreatic cancer and control tissues were identified in the GSE62452 and GSE28735 datasets,and an intersection analysis was conducted with the target-regulated mRNAs of the identified DEmiRs.Enrichment analysis was performed on DEmRs regulated by circBFAR target DEmiRs to identify genes involved in the PI3K-Akt signaling pathway,and survival curves were plotted.Ten pancreatic cancer tissues and corresponding adjacent normal tissues were collected,and RT-qPCR and Western blot were used to detect the expression of PI3K-Akt pathway genes.In the pancreatic cancer cell line PANC-1,circBFAR or hsa-miR-424-5p was knocked down,and the cells were divided into five groups for the experiments:control group,si-circNC group,si-circBFAR group,si-circBFAR+inhibitor NC group,and si-circBFAR+hsa-miR-424-5p inhibitor group.Cell viability was detected using the CCK-8 assay,cell migration and invasion were assessed by wound healing and Transwell assays,and the expression of signaling pathway genes was detected by RT-qPCR and Western blot.Results The expression level of circBFAR in pancreatic cancer tissues was significantly higher than that in the control group(P＜0.001).Among the identified target DEmiRs,hsa-miR-424-5p was downregulated in pancreatic cancer(P＜0.01).Target DEmRs regulated by hsa-miR-424-5p,including LAMA3,ITGA3,MET,and AREG,are involved in the PI3K-Akt signaling pathway,and their high expression in pancreatic cancer is associated with poor prognosis(P＜0.001).RT-qPCR and Western blot results showed that circBFAR,LAMA3,ITGA3,MET,and AREG were highly expressed in pancreatic cancer patients,while hsa-miR-424-5p was lowly expressed.Transfection of si-circBFAR in PANC-1 cells significantly inhibited cell proliferation,migration,and invasion,as well as the expression of LAMA3,ITGA3,MET,and AREG.Transfection of hsa-miR-424-5p inhibitor counteracted the effects of si-circBFAR on the cells(P＜0.01).Conclusion circBFAR promotes the progression of pancreatic cancer by regulating the expression of hsa-miR-424-5p and its target genes LAMA3,ITGA3,MET,and AREG in the PI3K-Akt signaling pathway.Inhibiting circBFAR may be a potential strategy for the treatment of pancreatic cancer.
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维普
