Molecular mechanism of regulating miR-223-3p/RHOB expression by circRNA SAMD8 to inhibit progression of pancreatic ductal adenocarcinoma
Objective To investigate the potential mechanism of circRNA SAMD8(circ-SAMD8)in development of pancreatic ductal adenocarcinoma(PDAC).Methods The expression profile of circRNAs in PDAC tissues was analyzed based on Microarray data(GSE79634).Quantita-tive real-time polymerase chain reaction(qRT-PCR)was used to verify the expression of circ-SAMD8(hsa_circ_0006148)in PDAC tissues and cells(CFPAC-1 and PANC-1).The target microRNA(miRNA)of circ-SAMD8 and its downstream mRNA were predicted by bioinformatics analysis,and identified by double luciferase reporter gene assay.The proliferation ability of PDAC cells was detec-ted by MTT assay and colony formation assay.The expression levels of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax were detected by Western blot.The percentage of apoptotic cells was detected by flow cytometry.Results The expression levels of circ-SAMD8 in PDAC tissues and cells were significantly lower than those in paracancer tissues and normal cells(P＜0.05).Overexpression of circ-SAMD8 could effectively promote apoptosis of PDAC cells and inhibited proliferation of PDAC cells.Dual luciferase reporter gene experiments showed that miR-223-3p was a potential interacting mol-ecule of circ-SAMD8,and the downstream mRNA target of miR-223-3p was RHOB.The miR-223-3p was abnormally overexpressed in PDAC tissues and cells,accompanied by low RHOB expression.In PDAC cells transfected with miR-223-3p mimics or sh-circ-SAMD8,RHOB expression was significantly decreased,proliferation ability was enhanced,and apoptosis rate was decreased.Conclusion Circ-SAMD8 regulates the expression of RHOB downstream by sponging miR-223-3p,and effectively inhibits the occurrence and development of PDAC.
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维普
