环状RNA circEIF4G2靶向微小RNA-379-5p诱导婴幼儿血管瘤内皮细胞增殖、迁移和侵袭的机制

Mechanism of circular RNA circEIF4G2 targeting microRNA-379-5p to induce the proliferation,migration and invasion of infantile hemangioma endothelial cells

目的:探讨circEIF4G2对婴幼儿血管瘤内皮细胞(HemECs)增殖、迁移和侵袭的影响及其机制。方法:收集2015年8月至2019至11月于山西医科大学第二医院行手术治疗的27例婴幼儿血管瘤患者的瘤组织和瘤旁正常皮肤组织,HemECs购自上海钦诚生物科技有限公司,实时定量聚合酶链反应(RT-qPCR)法检测组织和细胞中circEIF4G2和微小RNA-379-5p(miR-379-5p)表达。将HemECs分为si-NC、si-circEIF4G2、miR-379-5p、miR-NC、si-circEIF4G2+anti-miR-379-5p、si-circEIF4G2+anti-miR-NC组,采用细胞计数试剂盒(CCK-8)法检测各组细胞增殖,Transwell检测各组细胞迁移和侵袭。双荧光素酶报告基因实验验证circEIF4G2和miR-379-5p调控关系。两组间比较采用独立样本t检验;多组间比较采用单因素方差分析,进一步两组间比较采用LSD-t检验。结果:婴幼儿血管瘤组织中circEIF4G2表达高于正常皮肤组织(3.76±0.42比1.00±0.13,t=32.619,P<0.05),而miR-379-5p表达低于正常皮肤组织(0.46±0.07比1.00±0.12,t=20.197,P<0.05)。HemECs中circEIF4G2表达高于正常血管内皮细胞(2.61±0.20比1.00±0.07,t=22.794,P<0.05),而miR-379-5p表达低于正常血管内皮细胞(0.41±0.03比1.00±0.09,t=18.657,P<0.05)。si-circEIF4G2、si-NC组HemECs的吸光度(A)值分别为0.57±0.04、1.01±0.07,迁移数分别为(82.00±3.17)、(191.00±8.26)个,侵袭数分别为(61.00±3.11)、(132.00±7.73)个,si-circEIF4G2组各检测指标均低于si-NC组(t=16.187、36.960、25.564,P均<0.05)。miR-379-5p组和miR-NC组HemECs的A值分别为0.61±0.05、1.11±0.09,迁移数分别为(94.00±5.10)、(187.00±6.10)个,侵袭数分别为(71.00±4.27)、(138.00±8.10)个,miR-379-5p组各检测指标均低于miR-NC组(t=13.768、35.089、21.951,P均<0.05)。circEIF4G2靶向结合并负调控miR-379-5p表达。si-circEIF4G2+anti-miR-379-5p组和si-circEIF4G2+anti-miR-NC组HemECs的A值分别为1.04±0.09、0.58±0.05,迁移数分别为(184.00±10.02)、(86.00±4.10)个,侵袭数分别为(143.00±8.06)、(65.00±2.16)个,si-circEIF4G2+anti-miR-379-5p组各检测指标均高于si-circEIF4G2+anti-miR-NC组(t=13.462、27.156、28.043,P均<0.05)。结论:circEIF4G2在血管瘤组织和HemECs中呈高表达,下调其表达可能通过靶向负调控miR-379-5p抑制HemECs增殖、迁移和侵袭,其可能成为婴幼儿血管瘤治疗的分子靶点。

Objective To investigate the effect of circEIF4G2 on the proliferation,migration and invasion of infantile hemangioma endothelial cells(HemECs)and its mechanism.Methods The tumor tissue and normal skin tissue adjacent to the tumor were collected from 27 cases of infantile hemangioma who underwent surgical treatment at the Second Hospital of Shanxi Medical University from August 2015 to November 2019.HemECs were cultured in vitro.The expression of circEIF4G2 and microRNA(miR)-379-5p in the tissues and cells was detected by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR).HemECs were divided into si-NC group,si-circEIF4G2 group,miR-379-5p group,miR-NC group,si-circEIF4G2+anti-miR-379-5p group and si-circEIF4G2+anti-miR-NC group.Cell counting kit-8(CCK-8)assay was used to detect the cell proliferation,and Transwell was used to detect the cell migration and invasion.The dual luciferase reporter gene experiment verified the regulatory relationship between circEIF4G2 and miR-379-5p.Results The expression of circEIF4G2 in infantile hemangioma tissues was significantly higher than that in normal skin tissue(3.76±0.42 vs.1.00±0.13,t=32.619,P<0.05),but the expression of miR-379-5p was significantly lower than that in normal skin tissue(0.46±0.07 vs.1.00±0.12,t=20.197,P<0.05).As compared with the normal vascular endothelial cells,the expression of circEIF4G2 in HemECs was significantly increased(2.61±0.20 vs.1.00±0.07,t=22.794,P<0.05),but the expression of miR-379-5p was significantly reduced(0.41±0.03 vs.1.00±0.09,t=18.657,P<0.05).The A values of HemECs in the si-circEIF4G2 group and si-NC group were 0.57±0.04 and 1.01±0.07,the number of migrating cells was(82.00±3.17)and(191.00±8.26),and the number of invasive cells was(61.00±3.11)and(132.00±7.73),respectively.The detection indexes in si-circEIF4G2 group were significantly lower than those in the si-NC group(t=16.187,36.960,25.564,P all<0.05).The A values of HemECs in the miR-379-5p group and miR-NC group were 0.61±0.05 and 1.11±0.09,the number of migrating cells was(94.00±5.10)and(187.00±6.10),and the number of invasive cells was(71.00±4.27)and(138.00±8.10),respectively.The detection indexes in miR-379-5p group were significantly lower than those in miR-NC group(t=13.768,35.089,21.951,P all<0.05).The circEIF4G2 could negatively regulate the expression of miR-379-5p.The A values of HemECs in the si-circEIF4G2+anti-miR-379-5p group and si-circEIF4G2+anti-miR-NC group were 1.04±0.09 and 0.58±0.05,the number of migrating cells was(184.00±10.02)and(86.00±4.10),and the number of invasive cells was(143.00±8.06)and(65.00±2.16)respectively.The detection indexes in the si-circEIF4G2+anti-miR-379-5p group were significantly higher than those in the si-circEIF4G2+anti-miR-NC group(t=13.462,27.156,28.043,P all<0.05).Conclusion circEIF4G2 were highly expressed in hemangioma tissues and HemECs,and down-regulating its expression may inhibit the proliferation,migration and invasion of HemECs by targeting and negatively regulating miR-379-5p.It may become a molecular target for the treatment of infantile hemangioma.