Identification and Cloning of an inhibitor of Cancer Metastasis and Invasion

癌症转移和侵袭抑制剂的鉴定和克隆

• 批准号: 08671882
• 负责人: KIKKAWA Fumitaka
• 金额: $ 1.41万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08671882/
• 关键词: Ovarian Cancer; Metastasis; Fibronectin; MMP; TIMP; レチノイン酸

Conditioned media of normal peritoneal, myometrial, and ovarian tissues increased MMP-2 and -9 secretion from ovarian cancer (NOM1) cells in a concentration dependent manner. Especially, MMP-9 was markedly augmented by addition of normal peritoneal tissue conditioned medium. To estimate MMP-9 stimulating factors, conditioned medium of normal peritoneal tissue was fractionated and the influence of each fraction on MMP-9 secretion was examined by zymography. More than 100 kD fraction increased mostly MMP-9 secretion from NOM1 cells. Extracellular matrix, laminine, fibronectin, and type IV collagen, were added to NOM1 cells to identify the more than 100 kD factors. Fibronectin could increase MMP-9 secretion from NOM1 cells in a concentration dependent manner. Furthermore, fibronectin induced secretion of MMP-9 was inhibited by pretreatment with RGD peptides (fibronectin receptor) and antibody against integrin alpha5 receptor. Concentration of fibronectin was 164 ng/ml after 24 hours-culture of normal peritoneal tissue. Since 100 ng/ml fibronectin was enough to stimulate MMP-9 secretion from ovarian cancer cells, fibronectin was suggested to be an important stimulating factor.

正常腹膜，肌层组织和卵巢组织的条件培养基以浓度依赖性的方式增加了卵巢癌（NOM1）细胞的MMP -2和-9分泌。特别是，通过添加正常的腹膜组织条件培养基来显着增强MMP-9。为了估计MMP-9刺激因子，分离了正常腹膜组织的条件培养基，并通过Zymography检查了每个馏分对MMP-9分泌的影响。超过100 kD的分数大多增加了NOM1细胞的MMP-9分泌。将细胞外基质，椎板氨酸，纤连蛋白和IV型胶原蛋白添加到NOM1细胞中，以识别超过100 kD的因子。纤连蛋白可以以浓度依赖性方式增加NOM1细胞的MMP-9分泌。此外，用RGD肽（纤连蛋白受体）预处理纤连蛋白诱导的MMP-9分泌和抗整合素α5受体的抗体。正常腹膜组织培养24小时后，纤连蛋白的浓度为164 ng/ml。由于100 ng/ml纤连蛋白足以刺激卵巢癌细胞的MMP-9分泌，因此纤连蛋白被认为是重要的刺激因子。

项目成果

Koji Tamakoshi, Fumitaka Kikkawa, Osamu Maeda, Nobuhiko Suganuma N,Sadako Yamagata, Tatsuya Yamagata, Yutaka Tomoda: "Hyaluronidase activity in gynaecologic cancer tissues with different metastatic forms." Brit J Cancer. 75. 1807-1811 (1997)

Koji Tamakoshi、Fumitaka Kikkawa、Osamu Maeda、Nobuhiko Suganuma N、Sadako Yamagata、Tatsuya Yamagata、Yutaka Tomoda：“不同转移形式的妇科癌症组织中的透明质酸酶活性。”

Kiyosumi Shibata, Fumitaka Kikkaw, Koji Tamakoshi, Nobuhiko Suganuma, and Yutaka Tomoda: "Increased matrix metalloproteinase-9 activity in human ovarian cancer cells cultured with conditioned medium from human peritoneal tissue." Clin.Exp.Metastasis. 15.

Kiyosumi Shibata、Fumitaka Kikkaw、Koji Tamakoshi、Nobuhiko Suganuma 和 Yutaka Tomoda：“用来自人腹膜组织的条件培养基培养的人卵巢癌细胞中基质金属蛋白酶 9 活性增加。”

K.Tamakoshi: "Oifferent pattern of zymography between human gyecologic normal and malignant tissues." Am.J.Obstet.Gynecol.171. 478-484 (1994)

K.Tamakoshi：“人类妇科正常组织和恶性组织之间的酶谱分析模式不同。”

Kiyosumi Shibata, Fumitaka Kikkawa, Akihiro Nawa, Nobuhiko Suganuma, and Michinari Hamaguchi: "Fibronectin Secretion from Human Peritoneal Tissue Induces Mr 92,000 Type IV Collagenase Expression and Invation in Ovarian Cancer Cell Lines." Cancer Res.57. 5

Kiyosumi Shibata、Fumitaka Kikkawa、Akihiro Nawa、Nobuhiko Suganuma 和 Michinari Hamaguchi：“人类腹膜组织的纤连蛋白分泌诱导卵巢癌细胞系中的 Mr 92,000 IV 型胶原酶表达和侵袭。”

Koji Tamakoshi: "Hyaluronidaseactivity ingynecologic cancer tissues with different metastatic forms." Brit J Cancer. 75. 1807-1811 (1997)

Koji Tamakoshi：“具有不同转移形式的妇科癌症组织的透明质酸酶活性。”