Cytokine production by human lung cancer and its paracrine regulation

人肺癌细胞因子的产生及其旁分泌调节

• 批准号: 07670669
• 负责人: SONE Saburo
• 金额: $ 1.47万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670669/
• 关键词: Lung cancer; Cytokine; Paracrine; Macrophage; IL-10; MCP-1; IL-1ra; 肺胞マクロファージ; IL-6; MCAF

Th2 cytokines such as IL-4, IL-10, and IL-13, suppress pro-inflammatory cytokine production by monocytes/macrophages. Since monocyte chemoattractant protein-1 (MCP-1) is presumed to play an important role in monocyte recruitment and activation during inflammatory and immune responses, we examined here the effects of these Th2 cytokines on MCP-1 production by human blood monocytes and alvcolar macrophages (AM). Unstimulated highly purified blood monocytes did not produce MCP-1 spontaneously while LPS treatment induced the production of MCP-1 and its mRNA expression. All Th2 cytokines tested, suppressed LPS-induced MCP-1 production and its mRNA expression although the suppressive effect of IL-13 was weaker than those of IL-4 and IL-10. In contrast, IL-10 but neither IL-4 nor IL-13, induced unstimulated peripheral blood monocytes to produce biologically active MCP-1 protein within 4h, reaching a maximal level at 12h, IL-10-induced MCP-1 production was reduced by pre-treatment of IL-10 wit … More h anti-IL-10 Ab, negating the involvement of contaminated endotoxin. Moreover, IL-10 induced MCP-1 mRNA expression in unstimulated monocytes, independent of de novo protein synthesis. Furthermore, human AM produced MCP-1 spontaneously, and the production was inhibited by IL-4 or IL-13, but rather augmented by IL-10. Thses findings suggest that IL-10 regulates MCP-1 production by monocytes/macrophages in a different way from other Th2 cytokines such as IL-4 and IL-13, and contributes to host defense responses.We previously established novel metastasis model of human lung cancer sells in SCID mice depleted of NK cells (Yano et al., Int.J.Cancer, 1996). Human lung squamous cell carcinoma (RERF-LC-AI) cells formed metastases mainly in the liver and kidneys whereas small cell lung carcinoma (H69/VP) cells formed metastases mainly in systemic lymphnodes and liver in NK-cell depleted SCID mice. This study was conducted to explore the effect of local cytokine production on metastasis formation. To accumulate or activate macrophages by local cytokine production from metastatic cancer cells, we transduced human M-CSF-gene inserted into pRc/CMV-MCSF or human MCP-1-gene inserted into BCMGSNeo-MCAF to H69/VP and/or RERF-LC-AI cells, respectively.Cytokine-gene transduction had no effects on expression of surface antigens and proliferation. In vivo growth of s.c.injected M-CSF producing cells in SCID mice was slower when compared with their parent and mock-transduced cells. The number of metastatic colonies of MCSF-AI-9-18 and MCSF-AI-9-24 cells in liver, but not in kidneys, was significantly reduced. The number of lymph node metastases of MCSF-VP cells was also inhibited when compared with their parent or mock-transduced cells. Treatment of SCID mice depleted of NK cells with anti-h-M-CSF Ab promoted liver metastases of MCSF-AI-9-18 and MCSF-AI-9-24 cells. Systemic treatment with rh M-CSF (i.p.) had no effect on metastases of RERF-LC-AI cells. In contrast, MCP-1-gene transduction to H69/VP cells did not affect in vivo growth or metastatic potential of H69/VP cells. These findings suggest that antimetastatic effect of M-CSF may be organ specific and that presence of local M-CSF may have a therapeutic benefit to inhibit the metastases of human lung cancer. Less

Th2细胞因子如IL-4、IL-10和IL-13抑制单核/巨噬细胞产生促炎细胞因子。由于单核细胞趋化蛋白-1 （MCP-1）被认为在炎症和免疫反应中单核细胞募集和激活中起重要作用，我们在这里研究了这些Th2细胞因子对人类血液单核细胞和肺叶巨噬细胞（AM）产生MCP-1的影响。未受刺激的高纯度血液单核细胞不能自发产生MCP-1，而LPS处理诱导MCP-1的产生及其mRNA的表达。所有Th2细胞因子均抑制lps诱导的MCP-1的产生及其mRNA表达，但IL-13的抑制作用弱于IL-4和IL-10。相比之下，IL-10诱导未受刺激的外周血单核细胞在4h内产生具有生物活性的MCP-1蛋白，但IL-4和IL-13均未诱导，并在12h达到最大水平，IL-10预处理后，IL-10诱导的MCP-1蛋白的产生减少，从而消除了污染内毒素的参与。此外，IL-10在未刺激的单核细胞中诱导MCP-1 mRNA的表达，独立于从头蛋白合成。此外，人AM自发产生MCP-1， IL-4或IL-13抑制MCP-1的产生，而IL-10则增强MCP-1的产生。这些发现表明，IL-10调节单核/巨噬细胞产生MCP-1的方式与其他Th2细胞因子如IL-4和IL-13不同，并有助于宿主防御反应。我们之前在NK细胞缺失的SCID小鼠中建立了新的人肺癌转移模型（Yano et al., intj .）。癌症,1996)。在nk细胞缺失的SCID小鼠中，人肺鳞状细胞癌（RERF-LC-AI）细胞主要转移到肝脏和肾脏，而小细胞肺癌（H69/VP）细胞主要转移到全身淋巴结和肝脏。本研究旨在探讨局部细胞因子的产生对转移形成的影响。为了通过转移癌细胞产生的局部细胞因子积累或激活巨噬细胞，我们分别将插入pRc/CMV-MCSF的人m - csf基因或插入bcmgsno - mcaf的人mcp -1基因转导至H69/VP和/或ref - lc - ai细胞。细胞因子基因转导对表面抗原的表达和增殖无影响。与亲本细胞和模拟转导细胞相比，sc注射的M-CSF产生细胞在SCID小鼠体内的生长速度较慢。MCSF-AI-9-18和MCSF-AI-9-24细胞在肝脏中的转移菌落数量明显减少，而在肾脏中没有。与亲本细胞或模拟转导细胞相比，MCSF-VP细胞的淋巴结转移数量也受到抑制。用抗h- m - csf Ab治疗NK细胞缺失的SCID小鼠，促进MCSF-AI-9-18和MCSF-AI-9-24细胞的肝转移。rh - M-CSF （i.p.）系统治疗对rref - lc - ai细胞的转移没有影响。相比之下，mcp -1基因转导到H69/VP细胞并不影响H69/VP细胞的体内生长或转移潜能。这些发现提示M-CSF的抗转移作用可能是器官特异性的，局部M-CSF的存在可能具有抑制人肺癌转移的治疗益处。少

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