Impacts of CircASPH Targeting miR-28-5p/IGF-1R Axis on Proliferation,Migra-tion,and Invasion of Ovarian Granulosa Cells in Polycystic Ovary Syndrome
Objective:To explore the impacts of cyclic RNA asparagine hydroxylase(CircASPH)targeting the miR-28-5p/insulin-like growth factor 1 receptor(IGF-1R)axis on the proliferation,migration,and invasion of ovari-an granulosa cells in polycystic ovary syndrome(PCOS).Methods:Human ovarian granulosa cells KGN and COV434 were used as research subjects,the targeting relationship among CircASPH,miR-28-5p,and IGF-1R was confirmed through dual luciferase reporter gene experiments and pull down experiments.KGN and COV434 cells were grouped into si-NC group,si-ASPH group,si-ASPH+anti NC group,and si-ASPH+anti miR-28-5p group.The mRNA expression levels of CircASPH,miR-28-5p,and IGF-1R mRNA were detected by qRT-PCR,cell proliferation,migration,and invasion were detected by MTT assay,Edu staining,and transwell cell assay,respec-tively;and Western blot was applied to detect the expression of proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),vimentin,N-cadherin,E-cadherin,and IGF-1R proteins.Results:Bioinformatics a-nalysis and dual luciferase reporter gene experiments showed that CircASPH,IGF-1R and miR-28-5p had targe-ted binding sites.Compared with si-NC group,the expression level of CircASPH,OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin in si-ASPH group were decreased,while the expression level of miR-28-5p and E-cadherin protein were increased,and the differences were statisti-cally significant(P<0.05).Compared with the si-ASPH+anti-NC group,the expression level of miR-28-5p and E-cadherin in the si-ASPH+anti-miR-28-5p group were decreased,and the OD490 value,Edu positive cell rate,cell migration and invasion number,MMP-2,vimentin and N-cadherin were increased,the differences were statisti-cally significant(P<0.05).Conclusions:In KGN and COV434 cells,inhibiting the expression of CircASPH can inhibit the proliferation,migration,invasion,and epithelial-mesenchymal transition of ovarian granulosa cells by reg-ulating the miR-28-5p/IGF-1R axis,which may become a new target for the treatment of PCOS.
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