title: The effect of Vasohibin-1 expression and tumor-associated macrophages on the angiogenesis in vitro and in vivo.
abstract: Vasohibin-1 is an intrinsic inhibitor of angiogenesis induced by VEGF-A. However, there little is known about the relationship between Vasohibin-1 expression, angiogenesis, and tumor-associated macrophages (TAMs). Vasohibin-1 expression, VEGF-A expression, microvessel density (MVD) marked with CD34, and density of cells marked with CD68 were measured in 111 paraffin-embedded tissues of gastric cancer by immunohistochemistry. The length of tube forming structures of endothelial cells and mobility rate of gastric cancer cells in Matrigel were tested by three-dimensional live cell imaging system. The effect of TAMs on the tumor growth, MVD, and Vasohibin-1 expression was measured by nude mice tumor genesis assay in vivo. We found that high Vasohibin-1 protein expression correlated significantly with worse TNM stage (P = 0.002), metastatic lymph node (P = 0.014), distant metastasis (P = 0.022), overall survival (P < 0.001), and progression-free survival (P < 0.001) compared to those with low Vasohibin-1 expression. Vasohibin-1 protein expression had statistical correlation with the MVD (r = 0.860, P < 0.001), density of CD68(+) cells (r = 0.882, P < 0.001), and VEGF-A expression (r = 0.719, P < 0.001) in the gastric cancer tissues. Decreasing Vasohibin-1 expression with siRNA increased the length of tube forming structures of endothelial cells in co-culture with endothelial cells (EA-hy923), macrophages, and gastric cancers (Hs746T). Tumor volume (P = 0.001), Vasohibin-1 (P < 0.001), and VEGF-A (P < 0.001) expression in mice inoculated with AGS and THP (10:1) was significantly higher than that with AGS alone (P = 0.001). Vasohibin-1 protein expression had statistical correlation with VEGF expression (r = 0.786, P < 0.001) and MVD (r = 0.496, P = 0.014) in gastric xenografted tumor. Therefore, Vasohibin-1 might be a potential marker of worse prognosis and therapeutic target in gastric cancer. Vasohibin-1 might play an important role in the process of angiogenesis regulated by TAMs.
authors.affiliation: Department of Gastroenterological Surgery, Laboratory of Surgical Oncology, Peking University People's Hospital, Beijing, 100044, People's Republic of China. shenlong1977@163.com.
authors.forename: Zhanlong
authors.initials: Z
authors.lastname: Shen
authors.affiliation: Department of Gastroenterological Surgery, Laboratory of Surgical Oncology, Peking University People's Hospital, Beijing, 100044, People's Republic of China.
authors.forename: Yichao
authors.initials: Y
authors.lastname: Yan
authors.affiliation: Department of Gastroenterological Surgery, Laboratory of Surgical Oncology, Peking University People's Hospital, Beijing, 100044, People's Republic of China.
authors.forename: Chunxiang
authors.initials: C
authors.lastname: Ye
authors.affiliation: Department of Gastroenterological Surgery, Laboratory of Surgical Oncology, Peking University People's Hospital, Beijing, 100044, People's Republic of China.
authors.forename: Bo
authors.initials: B
authors.lastname: Wang
authors.affiliation: Department of Gastroenterological Surgery, Laboratory of Surgical Oncology, Peking University People's Hospital, Beijing, 100044, People's Republic of China.
authors.forename: Kewei
authors.initials: K
authors.lastname: Jiang
authors.affiliation: Department of Gastroenterological Surgery, Laboratory of Surgical Oncology, Peking University People's Hospital, Beijing, 100044, People's Republic of China.
authors.forename: Yingjiang
authors.initials: Y
authors.lastname: Ye
authors.affiliation: Department of Surgery, Helsinki University Central Hospital, University of Helsinki, Helsinki, 00290, Finland.
authors.forename: Harri
authors.initials: H
authors.lastname: Mustonen
authors.affiliation: Department of Surgery, Helsinki University Central Hospital, University of Helsinki, Helsinki, 00290, Finland. Pauli.Puolakkainen@hus.fi.
authors.forename: Pauli
authors.initials: P
authors.lastname: Puolakkainen
authors.affiliation: Department of Gastroenterological Surgery, Laboratory of Surgical Oncology, Peking University People's Hospital, Beijing, 100044, People's Republic of China. shanwang60@sina.com.
authors.forename: Shan
authors.initials: S
authors.lastname: Wang
mesh_terms: D000368:Aged; D000818:Animals; D002277:Carcinoma / Q000098:blood supply / Q000235:genetics* / Q000473:pathology; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D018572:Disease-Free Survival; D005260:Female; D064593:Heterografts; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008264:Macrophages / Q000502:physiology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D055806:Microvessels / Q000473:pathology; D008875:Middle Aged; D009362:Neoplasm Metastasis; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D009389:Neovascularization, Pathologic / Q000235:genetics / Q000503:physiopathology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D011897:Random Allocation; D016037:Single-Blind Method; D013047:Specific Pathogen-Free Organisms; D013274:Stomach Neoplasms / Q000098:blood supply / Q000235:genetics* / Q000473:pathology; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis
chemical_list: D018797:Cell Cycle Proteins; D009363:Neoplasm Proteins; D034741:RNA, Small Interfering; C493181:VASH1 protein, human; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A
keywords: Angiogenesis; Gastric cancer; Prognosis; TAMs; Vasohibin-1