Matrix-degrading Proteinases and their inhibitors in invasive and metastatic ovarian carcinomas.

侵袭性和转移性卵巢癌中的基质降解蛋白酶及其抑制剂。

• 批准号: 07671811
• 负责人: HIRAHARA Fumiki
• 金额: $ 1.47万
• 依托单位: YOKOHAMA CITY UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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-07671811/
• 关键词: OVARIAN NEOPLASMS; INVASION AND METASTASIS; PROTEINASES; PROTEINASE INHIBIOTORS; GENE EXPRESSION; トリプシン; トリプシノーゲン

<Objective> To elucidate themechanisms of invasion and metastasis of human ovarian carcinomas, we studied matrix-metalloproteinases, such as gelatinase A and B (type IV collagenase), andmultiple serine proteinases in vivo and in vitro. <Study design> Human ovarian carcinoma cell lines, transplanted tumor, and surgical samples obtained during operation, were employed for the study. Distribution of matrix-metalloproteinases and serine proteinases in ovarian carcinomas were analyzed by gelatin zymography. Their inhibitors were examined by reverse zymography. These proteinases and inhibitors were identified by immunoblotting methods. Trypsin (ogen) expression in ovarian carcinomas was determined by Northern blot analysis and immunohistochemical studies. <Results> By zymographic and Western blot analyzes, ovarian adenocarcinoma cell lines secreted multiple types of serine proteinases such as tissue-type and urokinase-type plasminogen activators, while undifferentiated adenocarcinomas mainly … More secreted gelatinase A or B.The high proteinase producers hardly secreted their corresponding inhibitors, such as tissue inhibitor of metalloproteinases (TIMP)-1, -2 or plasminogen activator inhibitor-1 (PAI-1). As for clinical samples, we showed high gelatinase activity in the cyst fluids derived from high grade human ovarian adenocarcinomas. When the ovarian carcinoma cell lines formed cystic xenoplant tumors in nude mice, high levels of these proteinase activities were demonstrated in the cystfluid. These high levels of gelatinase activities in transplanted cystic tumors were derived from the interaction of the tumor cells and surrounding interstitial fibroblasts. Trypsinogen mRNA and immunohistochemical stainings were detected in all of advanced ovarian adenocarcinomas, whereas it was undetectable in early stage carcinoma, low malignant potential and benign ovarian tumors as well as normal ovarian tissues. Especially, all of the serous cystadenocarcinmas tested showed high expression of trypsinogen mRNA compared to mucinous cystadenocarcinomas and clear cell adenocarcinomas at advanced stages. I<Conclusion> The present study suggests that the balance between the proteinases and their inhibitors might determine the malignant potential of the ovarian tumor cells to degrade matrix proteins and their capability of invasion and metastatic behavior. These data also strongly suggest that tumor-derived trypsin may implicate in the invasive and metastatic behavior of ovarian malignant tumors. Less

<Objective>为了阐明卵巢癌侵袭和转移的机制，我们在体内和体外研究了基质金属蛋白酶，如明胶酶A和B（IV型胶原酶），以及多种丝氨酸蛋白酶。<Study design>人卵巢癌细胞系、移植肿瘤和手术期间获得的手术样品用于研究。应用明胶酶谱法分析基质金属蛋白酶和丝氨酸蛋白酶在卵巢癌中的分布。用反向酶谱法检测其抑制剂。这些蛋白酶和抑制剂通过免疫印迹方法鉴定。用北方印迹法和免疫组化法检测卵巢癌中胰蛋白酶原的表达。<Results>通过酶谱和Western blot分析，卵巢腺癌细胞系分泌多种类型的丝氨酸蛋白酶，如组织型和尿激酶型纤溶酶原激活剂，而未分化腺癌主要分泌丝氨酸蛋白酶。 ...更多信息 高蛋白酶产生菌几乎不分泌相应的抑制剂，如金属蛋白酶组织抑制剂（TIMP）-1、-2或纤溶酶原激活物抑制剂-1（派-1）。至于临床标本，我们发现高级别卵巢腺癌的囊液中明胶酶活性较高。当卵巢癌细胞系在裸鼠中形成囊性异种移植瘤时，在囊液中显示出高水平的这些蛋白酶活性。移植性囊性肿瘤中高水平的明胶酶活性来源于肿瘤细胞和周围间质成纤维细胞的相互作用。胰蛋白酶原mRNA和免疫组化染色在所有晚期卵巢腺癌中均有表达，而在早期癌、低恶性潜能卵巢肿瘤、良性卵巢肿瘤和正常卵巢组织中均未检测到。特别是，所有的浆液性囊腺癌测试显示高表达的胰蛋白酶原mRNA相比，粘液性囊腺癌和透明细胞腺癌在先进的阶段。本<Conclusion>研究提示，蛋白酶及其抑制剂之间的平衡可能决定卵巢肿瘤细胞降解基质蛋白的恶性潜能及其侵袭和转移行为的能力。这些数据也有力地表明，肿瘤源性胰蛋白酶可能与卵巢恶性肿瘤的侵袭和转移行为有关。少

项目成果

Hirahara F: "Differential expression of trypsin in human ovarian carc_1homg and Low malignant potential tumns" Gynecol Oncol. (in press). (1998)

Hirahara F：“胰蛋白酶在人卵巢癌中的差异表达_1homg 和低度恶性潜能肿瘤”Gynecol Oncol。

Kawano N: "Expression of gelatinaseA,Tissre inhibitor of metalloputeihax-2 methilysin and trypsihosen in Lung neoplasms" Ham Pathol. 28. 613-622 (1997)

Kawano N：“肺肿瘤中明胶酶 A、metaloputeihax-2 甲硫氨酸和胰蛋白酶的 Tissre 抑制剂的表达”Ham Pathol。

Hirahara F: "Trypsinogen expression of human ovauan carcinomas" Int.J.Cancer. 63. 176-181 (1995)

Hirahara F：“人类卵巢癌的胰蛋白酶原表达”Int.J.Cancer。

Hirahara F: "Differentiation expression of trypsonogen in human ovarian carcinomeand low malgnaut potential tumars" Gynecol.Oncol. (in press). (1998)

Hirahara F：“胰蛋白酶原在人卵巢癌和低恶性潜能肿瘤中的分化表达”Gynecol.Oncol。

Miyagi Y: "Assignment of human pps/TFPI2 gene to 7q22 by FISH and PCR-based human/rodents cell-hybrid mapping panel-anclysis" Genomics. 35. 267-268 (1996)

Miyagi Y：“通过 FISH 和基于 PCR 的人类/啮齿动物细胞杂交图谱分析将人类 pps/TFPI2 基因分配给 7q22”基因组学。