基于外泌体ZFAS1调控miR-150-5p/VEGFA轴探讨食道通结方抑制食管癌血管新生的机制研究

肿瘤血管新生是食管癌生长的必要条件。外泌体LncRNA ZFAS1能促进食管癌细胞的增殖和转移。我们前期研究表明，过表达ZFAS1可促进食管癌细胞分泌外泌体ZFAS1，外泌体ZFAS1能下调与其共培养的血管内皮细胞miR-150-5p的表达，上调VEGFA蛋白的表达，诱导肿瘤血管生成。中药食道通结方干预能使食管癌细胞分泌外泌体ZFAS1的表达降低，使血管内皮细胞miR-150-5p上调，VEGFA蛋白下调，令血管生成受抑。据此提出科学假说：食道通结方可通过外泌体ZFAS1调控miR-150-5p/VEGFA轴抑制食管癌血管新生。本项目拟采用外泌体-细胞共培养、免疫荧光、移植瘤模型等方法从食道通结方抑制外泌体ZFAS1分泌、抑制血管生成表型、调控miR-150-5p/VEGFA轴及VEGFA/VEGFR2通路三方面阐明该方抗食管癌血管生成的机制，为健脾理气化痰法治疗食管癌提供新的实验依据。

Tumor angiogenesis is a necessary condition for the growth of esophageal cancer. Exosomal LncRNA ZFAS1 can promote the proliferation and metastasis of esophageal cancer cells. Our previous studies have shown that overexpression of ZFAS1 can promote tumor angiogenesis by promoting secretion of esophageal cancer cell derived exosomal ZFAS1. Exosomal ZFAS1 can downregulate the expression of miR-150-5p, upregulate the expression of VEGFA protein in vascular endothelial cells in the co-cultured system, thus inducing tumor angiogenesis. The traditional Chinese medicine Shidao Tongjie Decoction can inhibit tumor angiogenesis by reducing the expression of ZFAS1 in esophageal cancer derived exosome, upregulating the expression of miR-150-5p, downregulating the expression of VEGFA protein in vascular endothelial cells. Based on this, a scientific hypothesis was put forward that the decoction could inhibit angiogenesis of esophageal cancer by regulating miR-150-5p/VEGFA axis through exosomal ZFAS1. Exosome-cell co-culture, immunofluorescence, xenograft model and other methods were to be used to elucidate the mechanism of Shidao Tongjie Decoction inhibiting angiogenesis of esophageal cancer in three aspects: inhibition of secretion of exosomal ZFAS1, inhibition of angiogenesis phenotype, regulation of miR-150-5p/VEGFA axis and VEGFA/VEGFR2 signal pathway. We hope to provide new experimental evidence for the treatment of esophageal cancer with tonifying spleen, regulating qi and eliminating phlegm methods.