MiR-199a-5p Inhibits Proliferation And Induces Apoptosis In Hemangioma Cells Through Targeting HIF1?

Objective To investigate the function of miR-199a-5p(miR-199a)in HAs progression.Methods We collected normal skin tissues(n=15),proliferating phase HAs(n=15)and involunting phase HAs(n=15),qRT-PCR analysis of expression level of miR-199 a and PCNA(Proliferating Cell nuclear A antigen).Spearman correlation analysis of association of miR-199 a and PCNA expression in proliferating phase HAs and involunting phase HAs.After HDEC and CRL-2586 EOMA cells were transfected with miR-199 a mimic or miR-199 a inhibitor,evaluation of the cell proliferation activity and analysis of the cell apoptosis index.We screened the target genes of miR-199 a by using the starBase v2.o for bioinformatic analysis.After HDEC and CRL-2586 EOMA cells were transfected with miR-199 a mimic or miR-199 a inhibitor,qRT-PCR and western blotting analysis the expression of HIF1?.PmirGLO report vector carrying wild type 3?-UTR or mutated 3?-UTR of HIF1? was co-transfected with miR-199 a mimic or inhibitor into HDEC and CRL-2586 EOMA cells.Luciferase activities were examined with a Dual-luciferase Reporter System.Results(1)The expression level of miR-199 a was decreased,while that of PCNA was increased in proliferating phase HAs compare with the normal skin tissues and the involuting phase HAs.In addition,Spearman correlation analysis indicated that miR-199 a had a negative correlation with PCNA expresstion in proliferating phase HAs,but had no correlation in involuting phase HAs.(2)miR-199 a mimic suppressed cell proliferation activity and induced cell apoptosis in HDEC and CRL-2586 EOMA cells.The expression levels of PCNA and cleaved caspase-3 were detected by western blotting,indicating a decreased expression of PCNA and increased expression of cleaved caspased-3 caused by miR-199 a mimic.The results of miR-199 a inhibitor is the opposite.(3)After bioinformatic analysis,HIF1? ranked the first place and was considered to have the greatest potential to bind with miR-199 a.Besides,qRT-PCR and western blotting analysis showed that miR-199 a mimic decreased the expression of HIF1?,whereas miR-199 a inhibitor increased its expression in HDEC and CRL-2586 EOMA cells.The results of Dual-luciferaseReporter System illustrated that miR-199 a mimic reduced the luciferase activity of WT 3`UTR of HIF1?,while miR-199 a inhibitor increased this effect,but both of them had no effect on the luciferase activity of mutation 3` UTR of HIF1?.Conclusion(1)miR-199 a may inhibited HAs cell proliferation.(2)miR-199 a mimic inhibited cell proliferation and induced apoptosis,but miR-199 a inhibitor promoted proliferation and prevented apoptosis.(3)HIF1? was identified as a direct target of miR-199a.