Analysis of the function of Aminopeptidase N and neutral and peptidase and application of its cancer therapy.

氨肽酶N和中性肽酶的功能分析及其在癌症治疗中的应用。

• 批准号: 14571169
• 负责人: TOKUHARA Takahiro
• 金额: $ 1.86万
• 依托单位: The Tazuke Kofukai
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571169/
• 关键词: angiogenesis; APN; CD13; NEP; CD10

We have established the monoclonal antibody(MAb) MH8-11 suppressing angiogenesis and the cloning of this antibody reveals Aminopeptidase N(APN)/CD13. The APN transfected B16-F1 gained a potential ability of metastasis to the lung. Using the actual clinical specimens of lung cancer, colon cancer and pancreatic cancer, we found that the expression of APN had an association with angiogenesis, and was a significant factor of poor prognosis. On the other hand, we suppose neutral endopeptidase (NEP)/CD10 has anything to do with APN, but the evaluation of clinical specimens showed that NEP is a significant factor for good prognosis. MH7-5 MAb which we have established also recognized APN. Western blotting with MH7-5 showed both l6OkDa and l2OkDa bands, but only 150kDa band was detected using NH8-11. The modification of the sugar chain resulted in such changes, thus currently it is studied which kinds of sugar chain changes might associate with the tumor metastasis. One of our aims was to identify regulator genes of APN gene, and it was only observed that expression of NEP was decreased in the NEP positive and APN negative cell lines. Therefore, the identification of the gene which regulates APN gene remains unknown. Using the F1cell line with highly metastatic potential to the lung, we investigated whether the anti-APN MAb could prevent the lung metastasis or not. The number of metastatic colony of MH8-11MAb injected group were 41.4±24.4, compared to 98.3ア41.8 in control. This experiment demonstrated that MH8-11MAb efficiently prevented the pulmonary metastasis of F1 (p=0.0043). Currently, as the molecular targeting therapy, we will investigate whether human MH8-11 MAb might suppress the cancer progression or metastasis. We also investigate whether the NEP itself has a suppression of progression or not.

我们已经建立了抑制血管生成的单克隆抗体（MAb）MH 8 -11，该抗体的克隆揭示了氨肽酶N（APN）/CD 13。APN转染的B16-F1细胞具有潜在的肺转移能力。应用肺癌、结肠癌和胰腺癌的实际临床标本，我们发现APN的表达与血管生成有关，是预后不良的重要因素。另一方面，我们认为中性内肽酶（NEP）/CD 10与APN有关，但对临床标本的评估表明，NEP是预后良好的重要因素。我们建立的MH 7 -5单克隆抗体也识别APN。MH 7 -5蛋白质印迹显示160 kDa和120 kDa两条带，而NH 8 -11蛋白质印迹仅检测到150 kDa条带。糖链的改变导致了这些变化，因此目前正在研究哪些糖链的改变可能与肿瘤转移有关。我们的目的之一是鉴定APN基因的调控基因，并且仅观察到NEP阳性和APN阴性细胞系中NEP的表达降低。因此，APN基因的调控基因的鉴定仍然是未知的。本研究以F1细胞为实验对象，观察抗APN单克隆抗体对肺转移的抑制作用。MH 8 - 11单抗组的转移集落数为41.4±24.4，对照组为98.3 ± 41.8。该实验证明MH 8 - 11 MAb有效地防止F1的肺转移（p=0.0043）。目前，作为分子靶向治疗，我们将研究人MH 8 -11单克隆抗体是否可以抑制癌症的进展或转移。我们还调查了NEP本身是否具有进展抑制作用。

项目成果

Kim, KK., Flaherty, KR., Long, Q., Hattori, N., Sisson, TH., Colby, TV., Travis, WD., Martinex, FJ., Murray, S., Simon, RH.: "A plasnimogen activator inhibitor-1 promoter polymorphism and Idiopathic interstitial pneumonia."Mol Med. 9. 52-56 (2003)

Kim, KK.、Flaherty, KR.、Long, Q.、Hattori, N.、Sisson, TH.、Colby, TV.、Travis, WD.、Martinex, FJ.、Murray, S.、Simon, RH.：

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