Study On Effects Of MiR-210-3p And EphrinA3 On Tumor Angiogenesis In Oral Squamous Cell Carcinoma

Background:The process of tumor angiogenesis,which results for the interaction of many kinds of angiogenic and antiangiogenic factors,is one of the most important rate-limiting steps in the process of tumorigenesis,and the mechanism behind angiogenesis is very complicated.Although anticancer drugs targeting key molecules in angiogenesis have achieved relatively reliable clinical efficacy,some tumors are still insensitive to drug therapy or recurrent after treatment,which not only shows the diversity and complexity of tumor angiogenesis mechanism,but also indicates that there is still a long way to go in the in-depth research on tumor angiogenesis.As one of the miRNA relating to the angiogenesis,miR-210-3p has been confirmed to participate in the angiogenesis progress.Individual studies have suggested that miR-210-3p is associated with the development of oral squamous cell carcinoma(OSCC),and it is not known whether miR-210-3p is associated with angiogenesis in OSCC.EphrinA3,a member of the Eph family of axon-directed molecules,is targeted by miR-210-3p,but its role in OSCC has not been reported.Exosomes,which can contain a large number of proteins,DNA,miRNA and other substances,can serve as an important medium for cell-cell communication.In recent years,exxosome has become a research hotspot.Tumor cells can change the gene expression,information pathway state and biological behavior of the surrounding cells by releasing exosomes and then being absorbed by the surrounding receptor cells,so as to change the tumor matrix to promote angiogenesis and tumor metastasis.By studying the relationship between miR-210-3p and EphrinA3 and their effect on OSCC and the way in which this effect acts,we can explore the angiogenesis mechanism of OSCC.Objective: To investigate the expression of miR-210-3p and EphrinA3 in OSCC and its role in tumor angiogenesis.Methods:OSCC tissues and JCT were collected.Expressions of miR-210-3p and EphrinA3 and their relationship with tumor microvessel density(MVD)was detected.Mimic and inhibitor of miR-210-3p was designed and synthesized to change the expression of mir-210-3p in CAL27 and HUVECs and to observe their effect on CAL27 and HUVECs.Exosomes in CAL27 conditional media(CM)were isolated to detect the presence of miR-210-3p and to observe its delivery from tumor cells to HUVECs.The database was searched to investigate the targeting relationship between miR-210-3p and EphrinA3.The expression of EphrinA3 in HUVECs was altered by si RNA and lentivirus to explore the effect of EphrinA3 on the tube-forming ability of HUVEC,and then the activation status of relevant signaling pathways was detected.Results:The expression level of miR-210-3p in tumor tissues was higher than that in JCT,and the expression level was proportional to MVD in the tissues.EphrinA3 was inversely expressed,with more expression in JCT than in tumor tissues and inversely proportional to MVD.By changing the expression of miR-210-3p in CAL27,we found that the migration and proliferation of CAL27 cells decreased after the expression of miR-210-3p was decreased,while the migration and proliferation of CAL27 cells increased after the expression of miR-210-3p was increased.CAL27 CM had the ability to promote HUVEC tubulization,and the expression of miR-210-3p in HUVECs cells increased after being stimulated by CAL27 CM.When we changed the expression of mir-210-3p in HUVECs,we found that the increased level of miR-210-3p promoted the migration,proliferation and tube formation ability of HUVECs into tubes,while the decreased level of mir-210-3p decreased the migration,proliferation and tube formation ability of HUVECs.The expression of miR-210-3p was detected in exosomes extracted from CAL27 supernatant,and these exosomes alone can cause the increase of miR-210-3p expression in HUVECs.After tracing miR-210-3p in CAL27 cells with green fluorescence,we found that tumor cells delivered miR-210-3p to HUVECs cells through exosomes,and then HUVECs internalized the miR-210-3p and changed its biological behavior accordingly.By searching the database,we found that EphrinA3 was the target gene of miR-210-3p,and the expression of EphrinA3 would be changed after the expression of miR-210-3p was changed.Using si RNA and lentivirus to alter the expression of EphrinA3 in HUVEC,it was found that when EphrinA3 expression was reduced,the cell migration and proliferation capacity was enhanced,and the phosphorylation ratio of AKT was increased.When EphrinA3 expression was reduced,the cell migration and proliferation capacity and AKT phosphorylation ratio were reduced.Conclusions:The expression changes of miR-210-3p and EphrinA3 may be related to the angiogenesis of OSCC.Tumor cells delivered miR-210-3p to HUVECs through exosomes,and then targeted EphrinA3 in HUVECs and reduced its expression,finally activating the PI3K/AKT pathway and promoting the tube formation of HUVECs.