Effect of protein C inhibitor on renal cell carcinogenesis and on tumor invasion, growth and metastasis

Protein C抑制剂对肾细胞癌发生及肿瘤侵袭、生长和转移的影响

• 批准号: 16591592
• 负责人: HAYASHI Tatsuya
• 金额: $ 2.18万
• 依托单位: Mie University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591592/
• 关键词: Protein C inhibitor; renal cell carcinoma; CpG methylation; breast cancer cell; serine protease inhibitor; invasion; metastasis; reactive site; DNAメチル化; 腎近位尿細管上皮細胞

Protein C inhibitor (PCI), a member of the serine protease inhibitor (serpin) family, regulates the anticoagulant protein C pathway and inhibits urinary plasminogen activator (uPA), a mediator of tumor cell invasion. We recently demonstrated that PCI expression is significantly decreased in renal carcinoma cells derived from renal proximal tubular epithelial cells, and that PCI suppresses in vitro Matrigel invasion of renal carcinoma cells by inhibiting uPA. Furthermore, we also suggested that decreased expression of PCI in renal cell carcinoma (RCC) is mainly caused by methylation of the promoter region of PCI gene. In the present study, we elucidated the detailed mechanism of decreased expression of PCI induced by PCI gene methylation in RCC, and evaluated the effect of human PCI and its inactive derivatives on tumor cell invasion in vitro, and on tumor growth and metastasis in vivo using human breast cancer (MDA-231) cells. Hydrogen bisulfite DNA sequence showed that four out of thi … More rteen CpG sequence located near transcription initiation site are specifically methylated in PCI gene of RCC. Luciferase reporter gene assay using methylated PCI gene fragment showed that transcriptional activity of PCI gene promoter fragment is decreased by methylation of these four CpG sequence near transcription initiation site. On the other hand, the in vitro invasiveness of MDA-231 cells expressing intact PCI (MDA-PCI) was significantly decreased as compared to MDA-231 cells expressing inactive R354APCI (MDA-R354APCI) or the N-terminal fragment of PCI (MDA-NTPCI). Further, in vivo growth and metastatic potential of MDA-PCI, MDA-R354APCI and MDA-NTPCI cells in severe combined immunodeficient (SCID) mice were significantly decreased as compared to MDA-Mock cells. Overall, these studies showed that, decreased PCI expression in RCC is mainly caused by specific CpG methlation of PCI gene promoter, and in addition to its reactive site-dependent action, PCI also regulates tumor growth and metastasis independently of its protease inhibitory activity. Less

蛋白C抑制剂（PCI）是丝氨酸蛋白酶抑制剂（serpin）家族的一员，调节抗凝蛋白C通路并抑制尿纤溶酶原激活剂（uPA），uPA是肿瘤细胞侵袭的介质。我们最近证明，PCI表达显着降低，在肾癌细胞来源于肾近端肾小管上皮细胞，PCI抑制体外基质胶浸润的肾癌细胞通过抑制uPA。此外，我们还认为PCI基因在肾细胞癌（RCC）中表达降低主要是由于PCI基因启动子区甲基化所致。在本研究中，我们阐明了PCI基因甲基化诱导的肾癌PCI表达降低的详细机制，并评估了人PCI及其非活性衍生物对体外肿瘤细胞侵袭的影响，以及使用人乳腺癌（MDA-231）细胞对体内肿瘤生长和转移的影响。亚硫酸氢盐DNA序列显示， ...更多信息 肾癌PCI基因中位于转录起始位点附近的13个CpG序列发生特异性甲基化。利用甲基化的PCI基因片段进行荧光素酶报告基因检测，结果表明，PCI基因启动子片段的转录活性被转录起始位点附近的这四个CpG序列的甲基化所降低。另一方面，与表达失活R354 APCI（MDA-R354 APCI）或PCI的N-末端片段（MDA-NTPCI）的MDA-231细胞相比，表达完整PCI（MDA-PCI）的MDA-231细胞的体外侵袭性显著降低。此外，与MDA-Mock细胞相比，MDA-PCI、MDA-R354 APCI和MDA-NTPCI细胞在严重联合免疫缺陷（SCID）小鼠中的体内生长和转移潜力显著降低。总之，这些研究表明，PCI在RCC中的表达降低主要是由PCI基因启动子的特异性CpG甲基化引起的，并且除了其反应位点依赖性作用之外，PCI还独立于其蛋白酶抑制活性调节肿瘤的生长和转移。少

项目成果

Decreased protein C activation in patients with fulminant hepatic failure.

暴发性肝衰竭患者的蛋白 C 激活减少。

• 发表时间: 2006
• 期刊: Scand J Gastroenterol 41
• 作者: S.Kuno;E.Mizuta;S.Yamasaki;I.Araki;Yamauchi M
• 通讯作者: Yamauchi M

Activated protein C inhibits bronchial hyperresponsiveness and Th2 cytokine expression in mice.

• DOI: 10.1182/blood-2003-06-1980
• 发表时间: 2004-03
• 期刊: Blood
• 影响因子: 20.3
• 作者: H. Yuda;Y. Adachi;O. Taguchi;E. Gabazza;O. Hataji;H. Fujimoto;S. Tamaki;K. Nishikubo;K. Fukudome-K.-Fuk

Activated protein C inhibits bronchia 1 hytperresponsiveness and Th2 cytokine expression in mice.

活化的蛋白 C 抑制小鼠支气管 1 过度反应性和 Th2 细胞因子表达。

• 发表时间: 2004
• 期刊: Blood 103
• 作者: Yuda H;et al.
• 通讯作者: et al.

ヒトプロテインCインヒビター遺伝子トランスジェニックマウス.

人蛋白C抑制剂基因转基因小鼠。

• 发表时间: 2006
• 期刊: 血栓と循環 13
• 作者: I Araki;S Du;M Kamiyama;Y Mikami;K Matsushita;M Komuro;Y Furuya;M Takeda;林 辰弥
• 通讯作者: 林 辰弥

Impairment of IL-12-dependent STAT4 nuclear translocation in a patient with recurrent mycobacterium avium infection.

鸟分枝杆菌反复感染患者的 IL-12 依赖性 STAT4 核转位受损。

• 发表时间: 2004
• 期刊: J Immunol 172
• 作者: Oyama T;Kagami Y et al.;Koji Suzuki;Esteban C Gabazza;Tatsuya Hayashi;Yuda H;Toyoda H
• 通讯作者: Toyoda H