The role of nitric oxide in proliferation and angiogenesis of renal cell cancer

一氧化氮在肾细胞癌增殖和血管生成中的作用

• 批准号: 09671615
• 负责人: SAITO Kazuhide
• 金额: $ 1.79万
• 依托单位: NIIGATA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671615/
• 关键词: RCC; tumor thrombus; NO; iNOS; caveolin-1; caveolin-3; Immunohistochemistry; RT-PCR; 一酸化窒素; NOSII; 転移; 一酸化窒素合成酵素; 免疫組織学; altemative splicing; 増殖能

Background : Nitric oxide (NO) is generated in mammalian tissue by the conversion of L-arginine to L-citrulline. The reaction is catalyzed by nitric oxide synthase (NOS). NO has been suggested to have a biphasic action on tumor with both antitumor and tumor promotor activity, of which role has been still controversial. Recent reports demonstrated that Caveolin-1 and caveolin-3 inhibited eNOS and iNOS activity in vitro. In this study, we investigated the role of iNOS, caveolin-1 and caveolin-3 in progression of renal cell cancer (RCC).Materials and Methods : Surgical specimens were obtained from 50 patients of RCC.Immunoperoxidase staining was performed using the streptavidin-biotin bridge technique with staining anti-iNOS, caveolin-1, and caveolin-3 monoclonal antibodies. Apoptotic cells were examined by TUNEL method and subset analysis of tumor infiltrating lymphocytes (TIL) was also performed. RNA was extracted from RCC cell lines, ACHN, A498, KRCY, KH39, Cakil and Caki2, iNOS, caveolin-1 and caveolin-3 mRNA expression was examined by RT-PCR method.Results : Weak iNOS expression was detected by immunohistochemical staining only in 2 cases (0.5%) out of 50 primary tumors and in 1 out of 5 distant metastasis, In contrast, 2 out of 4(50%) of tumor thrombi were highly positive for iNOS.In vitro study, caveolin-1 was generally expressed on all cell lines, however, caveolin-3 expression was significantly reduced in more potent infiltrating phenotype, ACHN and KRC/Y than the others.Discussion and Conclusion : NO is one of the major angiogenesis factors in human neoplasm. Our data suggest that iNOS expression of tumor thrombus site of RCC may lead to intravascular infiltration and tumor progression. Downregulation of caveolin-3, an inhibitor of iNOS, may also play an important role in RCC progression in vivo.

背景：通过L-精氨酸转化为L-citrulline，在哺乳动物组织中产生一氧化氮（NO）。该反应是通过一氧化氮合酶（NOS）催化的。没有建议没有抗肿瘤和肿瘤启动子活性对肿瘤具有双相作用，而这种作用仍然存在争议。最近的报道表明，小窝蛋白-1和小窝蛋白3在体外抑制eNOS和iNOS活性。在这项研究中，我们研究了INOS，小窝蛋白-1和小窝蛋白3在肾细胞癌（RCC）进展中的作用。材料和方法：从50例RCC的患者中获得了手术样本。使用链霉亲素蛋白 - 抗蛋白抗蛋白抗激素 - 抗蛋白抗元素，以及30-抗氧蛋白 - 抗原蛋白 - 抗蛋白酶，以及30-抗抗氧素桥接的RCC。抗体。还通过TUNEL方法检查了凋亡细胞，并进行了肿瘤浸润淋巴细胞（TIL）的子集分析。从RCC细胞系，A498，KRCY，KH39，CAKI和CAKI2，INOS，Caveolin-1和Caveolin-3 mRNA表达中提取RNA。通过RT-PCR方法研究了RT-PCR方法：仅通过2例（0.5％的0.5％），在50次的50次中，通过RT-PCR方法进行了弱iNOS表达。在4（50％）中，肿瘤血栓对iNOS高度呈阳性。在体外研究中，小窝蛋白-1通常在所有细胞系上表达，但是，在更有效的浸润表型，ACHN和KRC/Y中，小窝蛋白-3的表达显着降低，而不是其他人。我们的数据表明，RCC肿瘤血栓部位的iNOS表达可能导致血管内浸润和肿瘤进展。 Caveolin-3（一种iNOS抑制剂）的下调也可能在体内RCC进展中起重要作用。