Characterization of the rat preneoplastic cell induction with glutathione S-transferase P form, GST-P,as marker enzyme

用谷胱甘肽 S-转移酶 P 形式、GST-P 作为标记酶诱导大鼠肿瘤前细胞的表征

• 批准号: 07457579
• 负责人: SATOH Kimihiko
• 金额: $ 0.96万
• 依托单位: Hirosaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457579/
• 关键词: chemical carcinogenesis; tumor marker; glutathione S-transferase; isozyme; glutathione; detoxicating enzyme; グルタチオン S-トランスフェラーゼ; 薬物代謝酵素; グツタチオン S-トランスフェラーゼ; GST-P

An investigation of the physiological functions and gene expression of a preneoplastic marker enzyme GST-P and the induction of the preneoplastic cells have pointed following results :1.Various GST substrates were effectively GSH-conjugated non-enzymatically at high GSH concentrations, by which it was indicated that the preneoplastic cells and neoplastic are activated not only enzymatically but also non-enzymatically. (Ref.1)2.Antibodies specific for six oncogene products, c-JUN,c-FOS and others, were prepared by fusion protein technology. Immunohistochemical examination of the rat hepatic preneoplastic lesions have shown that the GST-P expression was not correlated well with the expression of the oncogene products. (Ref.2)3.In the rat preneoplastic foci inducible by peroxisome proliferators such as clofibrate, GST-P and enoyl-CoA hydrolase, a peroxisomal enzyme, are negative. The Alpha and Mu class GST levels were shown to be decreaced in the foci as well. (Ref.3)4.In the acute stage … More of iron-induced rat renal carcinogenesis, GST-P mRNA increase was observed as early as one or two hours after administration of ferric nitrilotriacetate (15 mg/Kg body wt) suggesting the GST-P omdictopm against the carcinogenic insult and/or oxidative stresses. (Ref.4)5.A monoclonal antibody to rat GSTP1-1 was prepared and tryptic peptide mapping of the antigen has shown up the C-terminal 198-208 peptide epitope. (Ref.7)6.In the Nrf2 gene knockouted mice, inducibilities of the four GST isoenzymes as well as DT-diaphorase were lost against the butylated hydroxyanisole feeding, where Nrf2 is a trans-acting factor of globin genes. The result indicates the factor is closely related with the gene expression of Phase II detoxifying enzymes including mouse Pi class GSTMII.(Ref.11)7.A physicochemical approach has revealed that the hydrophobic subsite (the H-site) of the Pi class enzymes of rat GST-P and human GSTP1-1 are very low as compared to Alpha and Mu class GSTs, suggesting that the Pi classes are selective for weak electrophiles such as acrolein and hydoxyalkenals. (manuscript submitted).In a meanwhile, host-defensive roles of Pi class GSTs and the neoplastic cells agasint carcinogenic insult will be made clear. Less

本文研究了癌前标志酶GST-P的生理功能、基因表达及对癌前细胞的诱导作用，结果表明：1.在高浓度GSH条件下，各种GST底物均能有效地与GSH非酶结合，表明癌前细胞和癌细胞不仅受到酶的激活，而且受到非酶的激活。（参考文献1）2.利用融合蛋白技术制备了c-JUN、c-FOS等6种癌基因产物的特异性抗体。免疫组化结果显示，GST-P表达与癌基因产物表达无明显相关性。（参考文献2）3.在由过氧化物酶体增殖物如氯贝特诱导的大鼠癌前病灶中，GST-P和烯酰辅酶A水解酶（一种过氧化物酶体酶）是阴性的。α和Mu类GST水平在病灶中也显示出降低。（参考文献3）4.急性期 ...更多信息 在铁诱发的大鼠肾癌模型中，GST-P mRNA在次氮基三乙酸铁（15 mg/Kg体重）给药后1 ~ 2 h即明显增加，提示GST-P具有抗致癌损伤和/或氧化应激的作用。（参考文献4）5.制备了抗大鼠GSTP 1 -1的单克隆抗体，胰蛋白酶肽图分析显示C-末端198-208肽表位。（参考文献7）6.在Nrf 2基因敲除的小鼠中，四种GST同工酶以及DT-心肌黄酶的诱导性对于丁基化羟基茴香醚喂养丧失，其中Nrf 2是珠蛋白基因的反式作用因子。结果表明，该因子与包括小鼠Pi类GSTMII在内的II相解毒酶的基因表达密切相关。（参考文献1 - 1）7.物理化学方法已经揭示，与α和Mu类GST相比，大鼠GST-P和人GSTP 1 -1的Pi类酶的疏水亚位点（H-位点）非常低，这表明Pi类对弱亲电体如丙烯醛和羟基烯醛具有选择性。同时，对Pi类GST和肿瘤细胞在致癌物损伤中的宿主防御作用进行了探讨。少

项目成果

Kimihiko Satoh: "The high non-enzymatic conjugation rates of some glutathione S-transferase substrates at high glutathione concentrations." Carcinogenesis. 16(4). 869-874 (1995)

Kimihiko Satoh：“一些谷胱甘肽 S-转移酶底物在高谷胱甘肽浓度下具有高非酶结合率。”

Atassi, M.Z.: "Whitney,and M.Oshima;Mapping of the antibody-binding regions on botulinum neurotoxin H-chain domain 855-1296 with anti-toxin antibodies from three host species." J.Protein Chemistry. 15(7). 691-700 (1996)

Atassi, M.Z.：“Whitney 和 M.Oshima；用来自三个宿主物种的抗毒素抗体绘制肉毒杆菌神经毒素 H 链结构域 855-1296 上的抗体结合区图谱。”

A.Fukuda, T.Osawa, H.Oda, S.Toyokuni, K.Satoh and K.Uchida: "Oxidative stress response in iron-induced renal carcinogenesis : Acute nephrotoxicity mediates the enhanced expression of glutathione S-transferase Yp isozyme" Arch.Biocem.Biophys. 329 (1). 39-4

A.Fukuda、T.Osawa、H.Oda、S.Toyokuni、K.Satoh 和 K.Uchida：“铁诱导的肾癌发生中的氧化应激反应：急性肾毒性介导谷胱甘肽 S-转移酶 Yp 同工酶的表达增强”Arch

Suzuki S.et al.: "Lack of correlated expression between the glutathione S-transferase P-form and the oncogene products c-Jun and c-Fos in rat tissues and preneoplastic hepatic foci" Carcinogenesis. 16. 567-571 (1995)

Suzuki S.et al.：“大鼠组织和癌前肝病灶中谷胱甘肽 S-转移酶 P 型与癌基因产物 c-Jun 和 c-Fos 之间缺乏相关表达”致癌作用。

Shoji Nishimura, Yoshihito Yokoyama, Hajime Nakano, Kimihiko Satoh, Hiroko Kano, Kiyomi Sato and Shigeki Tsuchida: "Decreased expression of glutathione S-transferases and increased fatty change in peroxisomal enzye-negative foci induced by clofibrate in r

Shoji Nishimura、Yoshihito Yokoyama、Hajime Nakano、Kimihiko Satoh、Hiroko Kano、Kiyomi Sato 和 Shigeki Tsuchida：“在 r 中，氯贝特诱导的过氧化物酶体酶阴性病灶中谷胱甘肽 S-转移酶的表达降低，脂肪变化增加