Mechanism of miR-23c regulation of MELK on proliferation and Apoptosis of prostate cancer cell
Objective To explore the target genes of miR-23c and its role in proliferation,invasion,and apoptosis of prostate cancer cell.Methods Intervention experiments were performed to verify that miR-23c regulates the expression of MELK,which was divided into four groups,i.e.,LNCaP cell line(Control group),miR-NC(NC group),miR-23c inhibitor group,and miR-23c mimic group,and the latter three were transfected,respectively;MELK was screened out as a downstream target gene of miR-23c by the prediction software and the effect of miR-23c on protein expression was detected.The optimal interfering plasmid MELK-shRNA was determined by screening targeting experiments.Formal experiments were performed to detect the expression level of MELK,which was divided into three groups,i.e.,LNCaP cell line(Control group),transfected with shRNA-NC(shRNA-NC group)and MELK-shRNA(MELK-shRNA group).PCR was performed to detect the expression of MELK;CCK-8 assay was performed to detect cell proliferation;Transwell assay was performed to detect cell invasion ability and flow cytometry was performed to detect the degree of apoptosis of LNCaP cells.Reversion experiments were performed after co-transfection of miR-23c to verify that miR-23c inhibited the proliferation and invasion of LNCaP cells by targeting MELK.Results Bioinformatics analysis and prediction suggested that the presence of a binding site between miR-23c and MELK,and targeting experiments indicated that the expression of MELK could be affected by regulating the expression of miR-23c;invasion experiments revealed that the number of cell proliferation in the MELK-shRNA group was significantly decreased compared with that in the Control group(P<0.001);apoptosis rates for each group detected by flow cytometry were(8.42±0.50)%,(9.28±0.66)%,and(37.55±0.88)%,and the apoptosis rate of cells in MELK-shRNA group was significantly increased(P<0.001).The luciferase assays detected that miR-23c reduced the luciferase intensity of MELK-3'UTR(WT)to 28%,suggesting that miR-23c could regulate the expression of MELK.Conclusion miR-23c could inhibit the proliferation and invasion of LNCaP cells and increase the level of apoptosis by targeting MELK.
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