Regulation of has-miR-215-5p on endometrial glandular epithelial cell proliferation and migration by inhibiting the expression of BLCAP in hypoxia
Objective:To explore the potential mechanism of endometrial glandular epithelial cell(EEC)proliferation and migration under hypoxia condition.Methods:RNA from EEC under hypoxia(l%oxygen concentration)and normoxia(21%oxygen concentration)conditions were extracted for RNA-sequence to analyze the differences in intracellular microRNA(miRNA)gene expression between the two status.Reverse transcription quantitative polymerase chain reaction(RT-qPCR)was used to identify differentially expressed miRNA(has-miR-215-5p).The target genes of has-miR-215-5p were analyzed through miRNA target gene database website(Target Scan).Western blot was used to detect the expression of bladder cancer-associated protein(BLCAP).Cell counting kit-8(CCK-8)was used to observe the proliferation of EEC and the scratch test was used to detect the migration ability of EEC under hypoxia and normoxia conditions.Results:The miRNA sequencing results showed that the expression of has-miR-215-5p in EEC was significantly increased under hypoxia conditions(P<0.001),and the RT-qPCR result was the same as the sequencing results.The miRNA Target Scan database was searched to analyze and predict that BLCAP was the downstream target gene of has-miR-215-5p.The results of dual-luciferase reporter assay showed that has-miR-215-5p binded to BLCAP-3'-UTR,and the binding activity disappeared after the mutation of the binding site.Western blot analysis confirmed that the expression of BLCAP protein in EEC was significantly decreased under hypoxia conditions(P<0.001).CCK-8 and scratch experiments showed that the proliferation ability of EEC was significantly reduced and the migration ability was significantly enhanced under hypoxia conditions(P<0.001).Conclusions:Under hypoxia conditions,has-miR-215-5p in EEC may inhibit the expression activity of BLCAP by binding to BLCAP-3'-UTR,resulting in decreased proliferation and enhanced migration of EEC.