COX-2 and CCR2 induced by CD40 ligand and MCP-1 are linked to VEGF production in endothelial cells.
Recent studies have reported that expression of MCP-1 and its receptor, CCR2; and CD40-CD40 ligand (CD40L) interaction on mesenchymal cells play important roles in tumor development. Studies have also connected MCP-1, CCR2, and CD40L to COX-2 expression. The aim of this study was to examine the effect of MCP-1/CCR2 and CD40-CD40L interaction on COX-2 and VEGF expression in endothelial cells. We also investigated the localization of these proteins in gastric cancer tissue. COX-2 and CCR2 levels were evaluated in CD40L-stimulated HUVECs by Western blot and real-time PCR. VEGF secreted in the culture media was quantified by ELISA. Localizations of MCP-1, CD40L, CD34, CD40 and CCR2 in 34 gastric cancer tissue specimens were evaluated by immunohistochemistry. CD40-CD40L interaction-induced COX-2 production and subsequently, upregulated COX-2 production contributed to elevated VEGF and CCR2 levels in CD40L-stimulated HUVECs. CD40L-stimulated VEGF production was COX-2 but not COX-1 dependent. RS-102895, a CCR2-specific antagonist, significantly reduced VEGF production in CD40L- and MCP-1-stimulated HUVECs. MCP-1 had a synergistic effect on COX-2, CCR2 and VEGF levels in CD40L-stimulated HUVECs. In gastric cancer tissue, there was significant correlation between microvessel density and scores for CD40L, MCP-1 and CCR2 protein expression. Thus, MCP-1 had a synergistic effect on COX-2 and CCR2 protein expression in CD40L-stimulated HUVECs and thereby stimulated VEGF production in these cells.
Futagami
Seiji
S
Division of Gastroenterology, Department of Internal Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 1138603, Japan. seiji-f@nms.ac.jp
Hiratsuka
Tetsuro
T
Shindo
Tomotaka
T
Hamamoto
Tatsuhiko
T
Tatsuguchi
Atsushi
A
Nobue
Ueki
U
Shinji
Yoko
Y
Suzuki
Kenji
K
Kusunoki
Masanori
M
Tanaka
Shu
S
Wada
Ken
K
Miyake
Kazumasa
K
Gudis
Katya
K
Tsukui
Taku
T
Sakamoto
Choitsu
C
D048588:Benzoxazines / Q000378:metabolism*; D019013:CD40 Antigens / Q000378:metabolism; D023201:CD40 Ligand / Q000378:metabolism*; D002460:Cell Line; D018932:Chemokine CCL2 / Q000378:metabolism*; D017077:Culture Media, Conditioned; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D008833:Microcirculation; D010880:Piperidines / Q000378:metabolism*; D054390:Receptors, CCR2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D013274:Stomach Neoplasms / Q000378:metabolism / Q000473:pathology; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D048588:Benzoxazines; C111835:CCL2 protein, human; C515334:CCR2 protein, human; D019013:CD40 Antigens; D018932:Chemokine CCL2; D017077:Culture Media, Conditioned; D010880:Piperidines; C573791:RS 102895; D054390:Receptors, CCR2; D042461:Vascular Endothelial Growth Factor A; D023201:CD40 Ligand; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human