中国肺癌杂志2024，Vol.27Issue(11)：805-814,10.DOI:10.3779/j.issn.1009-3419.2024.102.38
骨转移性LUAD细胞通过HGF/YAP信号通路促进血管生成的机制研究
Mechanism of Bone-metastatic LUAD Cells Promoting Angiogenesis Through HGF/YAP Signaling Pathway
摘要
Abstract
Background and objective The early stages of tumor bone metastasis are closely associated with changes in the vascular niche of the bone microenvironment,and abnormal angiogenesis accelerates tumor metastasis and progression.However,the effects of lung adenocarcinoma(LUAD)cells reprogrammed by the bone microenvironment on the vascular niche within the bone microenvironment and the underlying mechanisms remain unclear.This study investigates the effects and mechanisms of LUAD cells reprogrammed by the bone microenvironment on endothelial cells and angiogenesis,providing insights into the influence of tumor cells on the vascular niche within the bone microenvironment.Methods The culture media from bone-metastatic LUAD cell A549-GFP-LUC-BM3(BM3-CM)and A549-GFP-LUC(A549-CM)were separately applied to human umbilical vein endothelial cell(HUVEC).A colony formation assay,scratch assay,and tube forma-tion assay were conducted to evaluate the proliferation,migration,and angiogenesis of HUVEC.Gene set enrichment analysis(GSEA)was conducted to identify enriched pathways,while reverse transcription quantitative polymerase chain reaction(RT-qPCR)and enzyme linked immunosorbent assay(ELISA)were performed to quantify hepatocyte growth factor(HGF),a protein that plays a crucial role in angiogenesis.Furthermore,the pivotal function of HGF and its underlying molecular mechanisms have been substantiated through the utilisation of recombinant proteins,neutralising antibodies,pathway inhibi-tors,immunofluorescence staining,and Western blot.Results BM3-CM demonstrated a more pronounced impact on the proliferation,migration,and angiogenesis of HUVEC compared to A549-CM.Bioinformatics analysis,combined with in vitro experiment,demonstrated that the secretory protein HGF was significantly elevated in BM3 cells and BM3-CM(P＜0.05).The addition of HGF neutralizing antibodies to BM3-CM inhibited the promoting effect of BM3-CM on HUVEC(P＜0.05),while the addition of recombinant HGF to A549-CM reproduced that promoting effect of BM3-CM on HUVEC(P＜0.05).HGF can enhance the activation of YAP(Yes-associated protein)in HUVEC,and this promotion effect may be achieved by activating Src and activating YAP into the nucleus(P＜0.05),but this effect can be inhibited by HGF neutralizing antibodies(P＜0.05).Furthermore,the addition of recombinant HGF to A549-CM can recapitulate the YAP activation effect of BM3-CM in HUVEC(P＜0.05).Conclusion Bone microenvironment reprogrammed bone-metastatic LUAD cells BM3 promote the proliferation,migration,and angiogenesis of HUVEC through the HGF/YAP axis,potentially playing a significant role in the modifications of the vascular niche.关键词
骨转移肺腺癌细胞/血管生成/HGF/YAPKey words
Bone-metastatic LUAD cells/Angiogenesis/HGF/YAP引用本文复制引用
邓妍,仇荣,刘星羽,苏英洋,薛阳,杜玉珍..骨转移性LUAD细胞通过HGF/YAP信号通路促进血管生成的机制研究[J].中国肺癌杂志,2024,27(11):805-814,10.基金项目
本研究受国家自然科学基金项目(No.81974315)资助 This study was supported by the grant from National Natural Science Foundation of China(No.81974315)(to Yuzhen DU). （No.81974315）
