Abstract:Objective: To investigate the effect of miR-31-5p on gefitinib resistance and its mechanism. Methods: The targeted miRNAs of serine protease 8 (PRSS8) were predicted by the online tools of TargetScan, miRDB, mirDIP and Encyclopedia of RNA Interactomes (ENCORI), and the gefitinib resistance-related miRNA microarray data was searched and downloaded from the high-throughput sequencing of the Gene Expression Omnibus (GEO) database for screening differentially expressed miRNAs. The expressions of miR-31-5p in gefitinib-sensitive (PC9) and gefitinib-resistant (PC9/GR) non-small-cell lung cancer (NSCLC) cell were detected by real-time fluorescence quantitative PCR (RT-qPCR). PC9/GR cells were transfected to miR-31-5p NC group and miR-31-5p mimic group, the sensitivity was tested by cell counting kit-8 (CCK-8) assay. The expression of PRSS8, BCL-2-associated x (BAX), B-cell lymphoma 2 (BCL-2) was detected by Western blot (WB). Results: Bioinformatics analysis showed that miR-31-5p was a miRNA that could target PRSS8 and had differential expression in gefitinib-resistant cells. The results from RT-qPCR revealed that miR-31-5p levels were significantly reduced in PC9/GR cells compared to PC9 cells (P<0.0001), the proliferation rate of cell in miR-31-5p mimic group was significantly lower than that of NC group ( all P<0.0001 ). Additionally, the expression of BAX protein was significantly elevated in the miR-31-5p mimic group relative to the NC group (P<0.001), but the level of BCL-2 was significantly decreased (P<0.05). Further studies showed that PRSS8 mRNA (P<0.0001) and protein (P<0.05) were significantly down-regulated in miR-31-5p mimic group. Conclusion: The expression of MiR-31-5p was significantly down-regulated in PC9/GR cell, and it may partly reverse gefitinib resistance by down-regulating the expression of PRSS8.
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