MicroRNA-1246 targeting glycogen synthase kinase-3β promotes prostate cancer migration,invasion,and proliferation by activating Wnt/β-catenin signaling pathway
Objective To investigate the effect and molecular mechanism of microRNA(miR)-1246 on the migration,invasion and proliferation of prostate cancer(PCa)cells by targeting glycogen syn-thase kinase 3β(GSK3β).Methods MiRNA-Seq was used to detect the differentially expressed miRNAs in PCa tissues and adjacent tissues.The expression levels of miR-1246 in cancer tissues and serum of PCa patients were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).Bioin-formatics analysis,luciferase reporter gene experiment,qRT-PCR and Western blotting were used to ana-lyze the targeted regulation relationship between miR-1246 and GSK3β.Human PCa cells DU145 and LNCaP were transfected with miR-1246 mimics/negative control(NC mimics)or miR-1246 inhibitor/negative control(NC inhibitor),respectively.The proliferation ability of cells was detected by cell counting kit-8(CCK-8)method,and the migration and invasion ability of cells were detected by scratch test and Transwell migration test.Independent sample t test was used for comparison between the two groups.Results The re-sults of miRNA-Seq showed that 15 miRNAs were significantly up-regulated and 51 miRNAs were signifi-cantly down-regulated in PCa tissues compared with adjacent tissues.Among them,miR-1246 was signifi-cantly up-regulated in PCa tissues and serum(tissue:7.93±2.39 vs.1.00±0.16,t=10.070,P＜0.01;Serum:15.23±2.77 vs.1.01±0.09,t=17.770,P＜0.01).Bioinformatics analysis showed that miR-1246 and GSK3β had complementary nucleotide sequences.The luciferase activity of miR-1246 mim-ics+WT-GSK3β group was significantly lower than that of NC mimics+WT-GSK3β group(0.58±0.09 vs.1.00±0.14,t=8.128,P＜0.05).The results of Western blotting showed that the expression of GSK3β in miR-1246 inhibitor group was significantly higher than that in NC inhibitor group(2.43±0.29 vs.1.00±0.09,t=10.180,P＜0.01),and the GSK3β expression in miR-1246 mimics group was sig-nificantly lower than that in NC mimics group(0.51±0.07 vs.1.01±0.12,t=5.120,P＜0.05).In DU145 and LNCaP cells,the cell proliferation ability,cell migration rate and number of invasive cells in the miR-1246 mimics group were significantly increased as compared with those in the NC mimics group,and those in the miR-1246 inhibitor group were significantly reduced as compared with those in the NC in-hibitor group.Western blotting results showed that the expression of Wnt3a and β-catenin in miR-1246 in-hibitor group was lower than that in NC inhibitor group(Wnt3a:0.39±0.05 vs.1.00±0.12,t=6.620,P＜0.01;β-catenin:0.54±0.09 vs.0.98±0.10,t=6.032,P＜0.01).Conclusion MiR-1246 promotes the migration,invasion and proliferation of PCa cells by inhibiting GSK3β and activating Wnt/β-catenin signaling pathway.
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