The Functional Study Of CD93 As A New Antiangiogenic Target In Infantile Hemangioma

ObjectiveInfantile hemangioma(IH)is mainly characterized by increased vasculogenesis,and CD93 is a newly discovered pro-angiogenic factor that is highly expressed in a variety of malignant tumors and promotes vasculogenesis.This study aims to investigate whether CD93 promotes the occurrence and development of IH by promoting vasculogenesis,and whether CD93 polyclonal antibody can inhibit the vasculogenesis of hemangioma stem cells(Hem SCs),then evaluate whether CD93 can be used as a new target for IH anti-vasculogenesis therapy.Methods(1)The expression levels of CD93 in IH tissues and normal skin were detected by immunohistochemistry.(2)CD133+ cells and CD133-cells were screened by magnetic-activated cell sorting,and the expression levels of CD133 and CD34 were measured by q PCR to identify whether the selected CD133+ cells were Hemangioma-derived stem cells(Hem SCs).(3)The expression levels of CD93 in CD133+ cells and CD133-cells were measured by Western blot,and the vascular formation ability of the two groups of cells was compared by the in vitro matrigel angiogenesis assay.(4)The CD93 overexpressed Hem SCs and CD93 interfered Hem SCs were obtained by lentivirus transfection method,and the expression of CD93 in CD93 overexpressed Hem SCs and CD93 interfered Hem SCs was verified by Western blot and q PCR.(5)In vitro matrigel angiogenesis assay were performed on the cells of the CD93 overexpression group,the CD93 interference group and the negative control group.The total length(Tot.length),number of junctions(Nb Junctions)and number of meshs(Nb meshes)of vessels formed by the three groups of cells in vitro were compared to observe whether overexpression of CD93 promoted the ability of vasculogenesis and whether interference of CD93 inhibited the ability of vasculogenesis.furthermore,CD93 polyclonal antibody was used to interfere with the in vitro matrigel angiogenesis of CD93 overexpressed Hem SCs to observe whether CD93 antibody could inhibit the ability of vasculogenesis of CD93 overexpressed Hem SCs.Results(1)The expression levels of CD93 are higher in proliferating IH tissues than paraneoplastic skin tissues,suggesting that CD93 may participate in the occurrence and development of IH.(2)CD133+ and CD133-cells were successfully isolated by magnetic-activated cell sorting method,and the selected CD133+ cells were Hem SCs.(3)the expression levels of CD93 and the ability of in vitro matrigel angiogenesis of Hem SCs cells are higher than that of CD133-cells,therefore,Hem SCs were used in subsequent experiments.(4)The CD93 overexpressed Hem SCs and three groups of CD93 interfered Hem SCs strains are successfully obtained by lentivirus infection method.The results of WB and q PCR show that the expression levels of CD93 are significantly increased in the CD93 overexpression Hem SCs and significantly decreased in three groups of CD93 interfered Hem SCs,of which the expression levels of CD93 was lowest in interference group II was used for subsequent experiments.(5)Compared with the negative control group,the total length,number of junctions and number of meshs of vessels are significantly increased in the CD93 overexpression group(p=0.0249,p=0.0325,p=0.0108),while are decreased in the CD93 interfered group(p= 0.0294,p=0.0396,p=0.0288).moreover,CD93 polyclonal antibody inhibits the ability of vasculogenesis of the CD93 overexpression Hem SCs,and the total length,number of junctions and number of meshs of vessels are significantly decreased(p<0.05,p<0.001,p<0.001).ConclusionCD93 was highly expressed in IH tissues.The overexpression of CD93 could improve the ability of vasculogenesis in vitro,while the interference of CD93 could inhibit the ability of vasculogenesis.Further studies revealed that the CD93 antibody could inhibit the function of vasculogenesis of CD93 overexpressing Hem SCs,which suggests that CD93 is a potential antiangiogenic therapeutic target for IH.There are no CD93-related studies was reported in IH research,and this study is an original research.At present,there is still lack of specific therapeutic targets for IH in clinical practice.As a potential anti-vasculogenesis target for IH,CD93 is expected to improve the treatment status of IH and has the prospect of translational medicine.