利用斑马鱼模型研究FGF19通过促进血管生成影响鼻咽癌转移的机制

Nasopharyngeal carcinoma (NPC) has a high tendency to metastasis and metastasis is an important factor that might affect the prognosis of NPC. Previous studies have shown that tumor angiogenesis exerts an important role in NPC metastasis. In the present study, we found that FGF19 is highly expressed in NPC and related to the metastasis of the patients. Besides, FGF19 could affect the biological behavior of vascular endothelial cells in tumor microenvironment and promote the progress of angiogenesis. In view of this, FGF19 could promote NPC angiogenesis and be involved in NPC metastasis. In this study, firstly, immunohistochemistry and ELISA were used to detect the expression of FGF19 in NPC. The relationship between FGF19 expression with metastasis and NPC microvessel density(MVD) were analyzed statistically. Secondly, we used CCK8, Transwell, tube formation, nude mice subcutaneous angiogenesis assay to investigate the effects of FGF19 on angiogenesis. Angiogenesis was also observed in a zebrafish model. Furthermore, the molecular mechanism of FGF19 on angiogenesis was studied by gene editing and high-resolution in vivo imaging in a transgenic zebrafish model. Our study might help to find new biomarkers and potential therapeutic targets that reflect NPC metastasis. What’s more, it might provide a new scientific basis for the treatment of NPC.

鼻咽癌（NPC）具有高度转移性，转移是影响其预后的重要因素。研究报道，肿瘤血管生成与NPC转移密切相关。申请人前期研究及预实验发现FGF19在NPC中高表达，与患者的转移有关；FGF19可影响肿瘤微环境中血管内皮细胞的生物学行为，促进血管生成。据此提出：FGF19可通过促进NPC血管生成，参与NPC转移。本课题拟通过免疫组化、ELISA等方法检测NPC中FGF19的表达水平，并统计分析其表达与转移及NPC微血管密度的关系；采用CCK8、Transwell、小管形成、裸鼠皮下血管形成实验及斑马鱼体内模型分析FGF19对血管生成的影响；接着在转基因斑马鱼模型中，应用基因编辑和高分辨率活体成像等技术深入研究FGF19影响血管生成的分子机制。本研究有助于找到反映NPC转移的新的生物学标志物和潜在治疗靶向，为NPC精准治疗提供新的科学依据。

鼻咽癌（NPC）具有高度转移性，转移是影响其预后的重要因素。研究发现，肿瘤血管生成与NPC转移密切相关。FGF19，作为成纤维细胞生长因子家族的重要一员，可促进细胞的增殖及分化。本项目探究FGF19在鼻咽癌血管生成中的功能及相关机制。首先利用免疫组化及western blot检测NPC组织中FGF19的表达水平，发现FGF19在NPC组织中高表达，与患者的临床分期及淋巴结转移有关，且高表达的患者预后更差。接着通过体外实验（CCK8、细胞流式、transwell迁移实验、划痕实验）和斑马鱼转移模型发现FGF19可促进NPC细胞的增殖和迁移。我们通过免疫组化分析发现FGF19表达与NPC组织微血管密度(MVD)有关，并通过体外实验（CCK8、transwell迁移实验、划痕实验、tube formation）及体内实验（裸鼠皮下血管形成、斑马鱼模型）等证实FGF19可促进血管生成。为进一步探究FGF19促进血管生成的机制，我们通过co-IP证实FGF19可与Annexin A2（ANXA2）相互作用，影响其表达。研究表明，当ANXA2下调后，FGF19的促血管生成作用受到抑制。最后，通过ELISA检测血清中FGF19的表达水平，发现NPC患者血清中FGF19表达高于正常人，且有转移者表达更高。本研究表明：NPC患者组织及血清中高表达的FGF19可通过影响ANXA2的表达，促进NPC血管生成，继而影响NPC转移。本研究阐述了FGF19在NPC血管生成中的作用及潜在机制，为NPC早期诊断和预后评估提供依据。