title: Tumor-associated macrophages induce capillary morphogenesis of lymphatic endothelial cells derived from human gastric cancer.
abstract: Tumor lymphangiogenesis is a major prognostic indicator of gastric cancer. Tumor-induced inflammation has been shown to attract tumor-associated macrophages that affect lymphangiogenesis. However, detailed mechanisms of macrophage-induced lymphangiogenesis have not been elucidated. Here, we evaluated the interaction between tumor-associated macrophages and lymphatic endothelial cells (LECs) derived from lymph nodes (LNs) of human gastric cancer. Lymphatic endothelial cells were directly or indirectly cocultured with macrophages from healthy human blood, with or without the supernatant of the gastric cancer cell line, OCUM-12. We analyzed the effect of cancer pretreated macrophages and of macrophages from metastatic LNs of gastric cancer on LECs. We observed morphological changes of LECs in coculture and assessed the gene expression of possible lymphangiogenic molecules of macrophages and LECs after contact coculture, and of cancer pretreated macrophages, by quantitative RT-PCR. Specimens of metastatic LN of gastric cancer were immunofluorescently stained. We found that tubulogenesis of LECs was observed only in the contact coculture model. OCUM-12 cells promoted macrophage-induced tubulogenesis of LECs. Relative gene expression of MMP and adhesion molecules was significantly upregulated in both capillary-forming LECs and cocultured macrophages. Cancer pretreated macrophages upregulated lymphangiogenic factors including inflammatory cytokines, MMPs, adhesion molecules, and vascular endothelial growth factor-C. Blocking of intercellular adhesion molecule-1 and macrophage activation suppressed tubulogenesis of LECs. Immunohistochemistry showed macrophages localized around lymphatic vessels. Our results suggested that interaction between LECs and macrophages may be an important initial step of tumor lymphangiogenesis developing LN metastasis. Understanding of its mechanisms could be useful for future therapeutics of gastric cancer.
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Yukie
authors.initials: Y
authors.lastname: Tauchi
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Tanaka
authors.affiliation: Department of Genetic Disease Research, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Kanako
authors.initials: K
authors.lastname: Kumamoto
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Mao
authors.initials: M
authors.lastname: Tokumoto
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Chie
authors.initials: C
authors.lastname: Sakimura
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Katsunobu
authors.initials: K
authors.lastname: Sakurai
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Kimura
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Toyokawa
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Amano
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Naoshi
authors.initials: N
authors.lastname: Kubo
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Muguruma
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Masakazu
authors.initials: M
authors.lastname: Yashiro
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Maeda
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Masaichi
authors.initials: M
authors.lastname: Ohira
authors.affiliation: Department of Surgical Oncology, Osaka City University Graduate School of Medicine, Osaka, Japan.
authors.forename: Kosei
authors.initials: K
authors.lastname: Hirakawa
mesh_terms: D002196:Capillaries / Q000254:growth & development*; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000235:genetics; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000166:cytology*; D020869:Gene Expression Profiling; D006801:Humans; D007249:Inflammation; D018836:Inflammation Mediators; D042583:Lymphangiogenesis; D008207:Lymphatic Metastasis; D042601:Lymphatic Vessels; D008264:Macrophages / Q000378:metabolism*; D020782:Matrix Metalloproteinases / Q000235:genetics; D009024:Morphogenesis; D013274:Stomach Neoplasms / Q000473:pathology*; D015854:Up-Regulation; D042582:Vascular Endothelial Growth Factor C / Q000235:genetics
chemical_list: D015815:Cell Adhesion Molecules; D018836:Inflammation Mediators; D042582:Vascular Endothelial Growth Factor C; D020782:Matrix Metalloproteinases
keywords: Cancer-related inflammation; gastric cancer; lymphangiogenesis; macrophage; tumor microenvironment