血管生成拟态(VM)是一种全新的肿瘤血供模式，前期发现VM周围线性排列的程序性坏死细胞（LPPCN）可能是VM形成的空间基础，但其影响VM的机制尚不清楚。本研究拟在102例人乳腺癌中观察程序性坏死关键RIP1通路成员与LPPCN、VM和内皮依赖性血管的空间分布关系及病理意义；构建LPPCN体外研究模型和细胞共培养体系观察其对周围细胞的影响；蛋白质谱鉴定由LPPCN细胞膜上解离的蛋白，筛选影响VM形成和血管内皮迁移的分子；运用多重荧光、细胞转染、IP和转基因小鼠、whole mount染色、共聚焦三维图像重建等验证LPPCN通过诱导乳腺癌细胞EMT和程序性坏死，促进VM形成并与宿主血管连通的分子机制；在动物模型中评估程序性坏死抑制剂治疗具有VM乳腺癌的可行性。结果有望明确VM形成的空间基础，阐明LPPCN调控周围细胞生物学行为主动促进VM的机制，并为临床治疗有VM的肿瘤提供新靶点和实验依据。

Vasculogenic mimicry (VM), identified in 1999, is a new tumor blood supply pattern. However, the space basement of VM channel in the tumor microentertainment with a high pressure is not clear. Previously, we have found that there were some line pattern programmed necrosis cells (LPPCN) surrounding to VM channels. There is association in distribution and time-line between VM and LPPCN. In this study, we will investigate the expression of RIP1 signaling pathway, which is essential in necroptosis, in 102 breast cancers analyze their clinical significance and the correlation between them, LPPCN and VM. Furthermore, we will establish a LPPCN model in vitro and a coculture system to observe the effect of LPPCNs on breast cancer cells and HUVCs. nanoLC-MS/MS will be used to identify the shedding proteins from cell membrane of LPPCN cells through RIP1-activaed ADAMs. The proteins associated with VM formation will be identified. Moreover, 3D cell culture, multiple immunofluorescent in 3D cell culture system, transfection, IP, EGFP transgenic SCID mice, whole mount, confocal and three dimensional image reconstruction will be performed to validate the molecular mechanisms which LPPCN induce breast cancer cells EMT, necroptosis and endothelial cells migration to promote VM formation and connection with host microcirculation. The necroptosis inhibitors will be administrated in breast cancer-bearing mice to assess the efficiency of them in treating tumors with VM. The results will reveal the origin and space of tumor cells lining VM channels. It will deep insight of LPPCN cells inducing VM through regulating other cells’ biological behaviors , and it will provide more new molecular targets for treatment of tumor with VM.