ENO1+外泌体促进口腔癌淋巴转移的分子机制研究

Oral squamous cell carcinoma (OSCC) is prone to develop lymphatic metastasis, which is the most important risk factor in determining patient’s poor survival. To address the scientific question that how OSCC initiate and develop lymphatic metastasis, we perform the pilot study and propose the hypothesis. We hypothesize that cancer associated fibroblasts (CAFs) of OSCC could release ENO1 positive exosomes under hypoxia conditions. ENO1 positive exosomes may facilitate extracellular matrix degradation by active invadopodia, which may be regulated by PI3K/Akt pathway. We will further investigate the molecular mechanism with following aspects: Firstly, to investigate the correlation of ENO1 positive exosomes with OSCC lymphatic metastasis and invadopodia by using tumor and peripheral blood specimen of OSCC patients; Secondly, to investigate the mechanisms of hypoxia induced ENO1 positive exosomes released by OSCC CAFs in vitro and explore the effect of ENO1 positive exosomes on activation of invadopodia by using hypoxia culture and functional knock down assay; Finally, to further verify how CAFs derived ENO1 positive exosomes promote lymphatic metastasis in OSCC orthograft mice model and quantum dots in vivo tracking system. Our research might illustrate the novel molecular mechanism of lymphatic metastasis of OSCC, and develop new strategies against OSCC lymphatic metastasis.

口腔鳞状细胞癌（口腔癌）易发生淋巴转移且严重影响患者生存。本项目围绕“口腔癌淋巴结转移的分子机制”这一科学问题，结合前期研究基础，提出科学假说：“低氧环境下口腔癌肿瘤相关成纤维细胞（CAFs）释放ENO1+外泌体通过激活PI3K/AKT信号通路促进侵袭伪足的形成，从而促进口腔癌淋巴结转移”。拟从以下几方面开展研究: （1）利用人口腔癌组织及外周血标本明确ENO1+外泌体与口腔癌淋巴结转移及侵袭伪足的相关性；（2）体外通过低氧培养，功能沉默探索低氧环境促进CAFs源性ENO1+外泌体的释放及调控侵袭伪足形成的分子机制；（3）利用舌癌淋巴转移小鼠模型及量子点体内示踪探索ENO1+外泌体对口腔癌淋巴结转移及侵袭伪足形成的调控作用及机制。本项目有望明确口腔癌淋巴转移的新机制。

口腔鳞状细胞癌（口腔癌）易发生淋巴转移且严重影响患者生存。本项目围绕“口腔癌淋巴结转移的分子机制”这一科学问题，本项目主要研究细胞外囊泡及组织中ENO1及代谢相关蛋白在口腔鳞癌淋巴转移中的作用。系统性回顾了组织源性细胞外囊泡在肿瘤及非肿瘤性疾病中的研究现状及瓶颈。利用新鲜口腔鳞癌组织样本及荧光多色标记技术发现：上调的肿瘤浸润CD103+树突状细胞及间质组织驻留记忆细胞密度提示口腔鳞癌患者良好预后。比较了发生转移淋巴及未发生淋巴转移的的口腔鳞癌患者的颈清术后引流液中的细胞外囊泡的差异蛋白：发生淋巴转移的口腔鳞癌患者颈清术后引流液中的细胞外囊泡代谢相关蛋白(EHD2和CAVIN1)显著上调，可能与口腔鳞癌的淋巴转移密切相关。在口腔鳞癌组织样本及细胞中，证实：在口腔鳞癌中ApoC3可与ENO1结合并促进IL-8的释放通过STAT3信号通路抑制T细胞增殖，上述研究结果为ENO1及代谢相关蛋白介导的口腔鳞癌淋巴转移提供了理论基础。