Studies on Protease Inhibitor Isolated from Breast Cancer and Breast Cancer Cell Line

乳腺癌及乳腺癌细胞系蛋白酶抑制剂的研究

• 批准号: 60480518
• 负责人: NISHIKI Masayuki
• 金额: $ 0.64万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60480518/
• 关键词: Breast caucer; Normal mammary gland; Breast cancer cell line; Purification; 精製

1) TPIs in human breast cancer extracts were significantly higher than in normal mammary gland extracts.2) TPI was purified from both human breast cancer and human normal mammary gland extracts by papain-Sepharose affinity chromatography and Sephacryl S-200 gel filtration.Two kinds of TPIs (Iow-molecular weight TPI and high-molecular weight TPI) were purified from both tissues. Their molecular weights were 14,000 and 90,000,respectively as determined by gel filtration.3) Low-molecular weight (LMW-) TPI had higher specific activity than high-molecular weight (HMW-) TPI. In breast cancer tissue extracts,LMW-TPI was dominant,Contrarily,HMW-TPI was dominant in normal gland tissue extracts.4) Only HMW-TPI reacted with anti-urinary thiol protease inhibitor (UTPI) rabbit IgG by double immunodiffusion and immunoelectrophoresis.5) LMW-TPI inhibited papain competitively using S-2302 as substrate.Its km and ki were 1.3x <10^(-3)> M and 6.1x <10^(-8)> M, respectively.6) LMW-TPI was found to be stable to heat and pH variation.7) TPIs were also purified from the human breast cancer cell line (YMB-l) . Both TPIs which were extracted from cultured cells and released into the medium, were confirmed to be LMW-TPI.Breast cancer cells may have lower ability to produce HMW-TPI than normal mammary gland cells. The difference of antigenicity to anti-UTPI IgG between HMW-TPI andLMW-TPI may be useful for diagnosis in near future.

1)人乳腺癌提取物中tpi含量显著高于正常乳腺提取物。2)通过木瓜蛋白酶- sepharose亲和层析和Sephacryl S-200凝胶过滤，从人乳腺癌和人正常乳腺提取物中纯化TPI。从两种组织中分别纯化出两种TPI（低分子量TPI和高分子量TPI）。凝胶过滤测定的分子量分别为14000和90000。3)低分子量（LMW-） TPI比高分子量（HMW-） TPI具有更高的比活性。在乳腺癌组织提取物中以低分子量tpi为主，而在正常乳腺组织提取物中以低分子量tpi为主。4)通过双免疫扩散和免疫电泳，只有hw - tpi与抗尿硫醇蛋白酶抑制剂（UTPI）兔IgG发生反应。5) lw - tpi以S-2302为底物对木瓜蛋白酶具有竞争性抑制作用。其km和ki分别为1.3x <10^(-3)> M和6.1x <10^(-8)> M。6) LMW-TPI对热和pH变化具有较好的稳定性。7)从人乳腺癌细胞株（ymb - 1）中也纯化到了tpi。从培养细胞中提取并释放到培养基中，证实两种tpi均为LMW-TPI。乳腺癌细胞产生HMW-TPI的能力可能低于正常乳腺细胞。HMW-TPI与lmw - tpi在抗utpi IgG抗原性上的差异，可能在不久的将来有助于诊断。

项目成果

久代淳一,山根基,西亀正之,片岡健,吉岡伸吉郎,内藤正志,伊藤明弘,土肥雪彦: 第45回日本癌学会総会記事. 196 (1986)

Junichi Kuyo、Hajime Yamane、Masayuki Nishikame、Ken Kataoka、Shinkichiro Yoshioka、Masashi Naito、Akihiro Ito、Yukihiko Doi：日本癌症协会第 45 届年会文章 196 (1986)。

Tsuyoshi KATAOKA: "Cytochemical hormone receptor assay of breast Cancer by aspiration biopsy cytology" Proceedirg of Japanese Cancer Association, 45th Annual Meeting. 420 (1986)

Tsuyoshi KATAOKA：“通过抽吸活检细胞学对乳腺癌进行细胞化学激素受体测定”日本癌症协会第 45 届年会论文集。

Motoi YAMANE: "Studies on Thiol Protease Inhibitor Isolated from Human Breast Cancer Tissue" Hiroshima Journal of Medical Sciences. 36. 39-52 (1987)

Motoi YAMANE：“从人乳腺癌组织中分离出硫醇蛋白酶抑制剂的研究”广岛医学科学杂志。

MOTOI,YAMANE: Hiroshima Journal of Medical Science. 36. 39-52 (1987)

本井，山根：广岛医学科学杂志。

山根基,西亀正之、吉岡伸吉郎,久代淳一,片岡健,松山敏哉,伊藤明弘,土肥雪彦: 第45回日本癌学会総会記事. 420 (1986)

Hajime Yamane、Masayuki Nishikame、Shinkichiro Yoshioka、Junichi Kudai、Ken Kataoka、Toshiya Matsuyama、Akihiro Ito、Yukihiko Doi：日本癌症协会第 45 届年会的文章 420 (1986)。