基于HIF-1α—COX-2—VEGF信号通路的大肠癌血瘀证血管新生机制及活血化瘀中药的作用研究

缺氧是大肠癌等实体瘤基本的病理特征，缺氧诱导HIF-1α过表达是大肠癌血管新生的重要原因，其临床表现、病理变化与大肠癌"血瘀证"证候类型暗合，但两者是否存在关联尚无类似研究。我们前期研究发现，活血化瘀中药丹参及有效成分通过HIF-1α--COX-2途径下调VEGF表达抑制血管生成。我们由此提出"血瘀证促大肠癌血管新生"假说。本课题以人大肠癌裸鼠模型及肿瘤血管新生三维细胞缺氧模型为研究对象，构建HIF-1α慢病毒载体质粒、采用Bio-Plex悬液芯片、慢病毒载体、双报告基因质粒转染、基因转染、RNAi等技术方法，研究HIF-1α-COX-2-VEGF信号通路在大肠癌瘀血证血管新生中的作用，并观察活血化瘀中药丹参主要成分丹参酮IIA的干预作用。从Hypoxia-HIF-1α-COX-2-Angiogenesis信号转导角度揭示血瘀证促血管新生的机制，为活血化瘀中药防治大肠癌提供理论依据。

Hypoxia is the basic pathological characteristics of colorectal cancer and other solid tumors, HIF-1α overexpression that induced by hypoxia is the important factors in colorectal cancer angiogenesis, its clinical manifestations, pathological changes were coincide with syndromes of blood stasis of colorectal cancer, but both existence of the association is currently no similar studies. We have previously shown that active ingredients of Salvia Miltiorrhizae, a traditional chinese medicine that activating blood and dissolving stasis, could significantly downregulate VEGF expression via HIF-1α-COX-2 pathway, and restrain its microvascular formation. Therefore, we infer hypothesis that syndrome of blood stasis induced tumor angiogenesis. Then in this program, we use human colorectal cancer cell xenograft tumor model in nude mice and 3-D angiogenesis cells model as research objects, and Bio-Plex Assay technology, Lentiviral vector infection, plasmid transfection, dual-luciferase reporter assay, and Western blotting will be employed to investigate the effects of HIF-1α-COX-2-VEGF pathway in colorectal angiogenesis with syndrome of blood stasis. To observe the intervention effect of Tanshinone IIA, the active ingredients of Salvia Miltiorrhizae as a traditional Chinese Durg that activating blood and dissolving stasis. To demonstrate mechanism that syndrome of blood stasis induced tumor angiogenesis through Hypoxia-HIF-1α-COX-2-Angiogenesis signaling transduction, and to provide thinking of Traditional Chinese Durg that activating blood and dissolving stasis preventing and treating colorectal cancer.