医学分子生物学杂志2026，Vol.23Issue(1)：51-59,9.DOI:10.3870/j.issn.1672-8009.2026.01.007
EPHB4对骨肉瘤细胞恶性进展的调控作用及机制研究
Regulation of EPHB4 on Malignant Progression of Osteosarcoma Cells
摘要
Abstract
Objective To explore the effect of erythropoietin-producing hepatocyte receptor B4(EPHB4)on the malignant progression of osteosarcoma and its mechanism.Methods The expres-sion level of EPHB4 in U2OS,SaOS2-LM7,SaOS2,and 143B was detected by Western blotting.SaOS2-LM7 cells was divided into 4 groups:control group,si-NC group,si-EPHB4 group,and sEPHB4(soluble EPHB4 blocking antibody)group.Cell proliferation was detected by CCK-8 assay and EdU staining,cell apoptosis was detected by flow cytometry,cell invasion and migration were detected by Transwell assay and Wound healing assay.A SaOS2-LM7 cell line la-beled with red fluorescent protein(RFP)which was stably silenced of EPHB4 was constructed(EPHB4-si-RNA-RFP).Twenty-four tumor-bearing nude mice were divided into 2 groups:the con-trol-RFP group and the EPHB4-si-RNA-RFP group.The lung metastasis was detected 1-4 weeks later by using small animal in vivo imaging system.Immunohistochemistry was used to analyze the expres-sion of EPHB4,cluster of differentiation 33(CD33),CYCLIN D1,matrix metalloproteinase 9(MMP-9),MMP-14,and β-CATENIN in tumor tissues 4 weeks later.Results The expression level of EPHB4 in the SaOS2-LM7,SaOS2,and 143B was significantly higher than that in the U2OS(all P<0.05).Compared with those in the control group,the expression level of EPHB4,the proliferation activity,and the EdU positive rate of SaOS2-LM7 in the sEPHB4 group and the si-EPHB4 group were significantly decreased,the apoptosis rate was significantly increased,and the migration rate and invasion rate were significantly decreased(all P<0.05).In addition,the rela-tive expression levels of β-CATENIN,CYCLIN D1,MMP-9,and MMP-14 in the sEPHB4 group and the si-EPHB4 group were also significantly decreased when compared with those in the control group(all P<0.05).The lung metastasis at 3 weeks and 4 weeks was less in the EPHB4-siRNA-RFP group than in the control-RFP group(both P<0.05).The protein expression of EPHB4,CD33,CYCLIN D1,MMP-9,MMP-14,and β-CATENIN in the tumor tissues of the EPHB4-siR-NA-RFP group was down-regulated(all P<0.05).Conclusion Silencing EPHB4 significantly in-hibits the activation of the WNT/β-CATENIN signaling pathway in osteosarcoma cells,suppresses cell proliferation,migration,and invasion,and promotes cell apoptosis.关键词
肝配蛋白B4/骨肉瘤/迁移/侵袭/凋亡/WNT/β-CATENIN信号通路Key words
erythropoietin-producing hepatocyte receptor B4/osteosarcoma/migration/inva-sion/apoptosis/WNT/β-CATENIN signaling pathway分类
医药卫生引用本文复制引用
张飞,陆红祥,买买提克里木·吐松江,杨童磊,刘秋宏,许刚..EPHB4对骨肉瘤细胞恶性进展的调控作用及机制研究[J].医学分子生物学杂志,2026,23(1):51-59,9.基金项目
新疆军区总医院院内课题-喀喇昆仑人才基金拔尖项目(No.2022BJ001) This work was supported by a grant from the Internal Research Project of Xinjiang Military Region General Hospital-"Karakoram"Talent Fund Top-notch Project(No.2022BJ001) （No.2022BJ001）
