The molecular mechanism of protein kinase C inhibitors on anti-metastasis

蛋白激酶C抑制剂抗转移的分子机制

基本信息

批准号：
14572087
负责人：
NAKAMURA Kazuki
金额：
$ 1.41万
依托单位：
Mukogawa Women's University
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-14572087/
关键词：
protein kinase C inhibitors PKC412 B16 mouse melanoma cells chemoinvasion spontaneous metastatic model experimental metastatic model integrin β1 rottlerin matrix metalloproteinase integrinβ_1 rotterin novel PKC conventional PKC 血

项目摘要

We investigated the mechanism of protein kinase C(PKC) inhibitors on anti-metastasis. PKC412 (4'-N-benzoyl staurosporine), a conventional PKC(alpha, beta and gamma) inhibitor, reduced the ability of mouse malignant melanoma(B16-BL6) cells to form lung colonies in mice and reduced invasion of the extracellular matrix in vitro when pie-incubated with the cells for 1 hour. Further, PKC412(200mg/kg/day for 4 weeks, p.o.) significantly prolonged survival time in a spontaneous metastatic mouse model, produced by subcutaneous inoculation of B16-BL6 cells(1×10^6 cells) into the right footpad of C57BL/6Cr mice Mowed by surgical amputation of the primary tumor 2 weeks after tumor inoculation. To elucidate mechanisms of anti-invasive action for PKC412, we measured cell motility, matrix metalloproteinase(MMP) activity secreted from cells and expression of integrin beta 1 protein in cells. As a result, PKC412 decreased the expression of integrin beta 1 protein of B16-BL6 cells in a dose-dependent manner. However, PKC412 could not inhibit cell motility and MMP activity of the melanoma cells. These results suggest that PKC412 inhibits the process of invasion in the metastatic pathway of melanoma cells by attenuation of integrin beta 1 expression. Finally, we examined metastasis-related PKC isoforms using rottlerin, a specific PKC delta inhibitor. Since rottlerin could not block the hematogenic lung metastasis in mice using B16-BL6 cells, conventional PKC isoforms were considered to induce tumor metastasis, while the delta isoform did not.

我们研究了抗近距离抑制剂蛋白激酶C（PKC）抑制剂的机制。 PKC412（4'-N-N-苯并二孢孢），一种常规的PKC（Alpha，beta和Gamma）抑制剂，降低了小鼠恶性黑色素瘤（B16-BL6）细胞在小鼠中形成肺部的能力（B16-BL6）细胞在小鼠中形成肺部的能力，并降低了与植物细胞的入侵，可与植物细胞的入侵，以便于1个小时，以获得1个小时的细胞。 Further, PKC412(200mg/kg/day for 4 weeks, p.o.) significantly prolonged survival time in a sponsorous metastatic mouse model, produced by subcutaneous inoculation of B16-BL6 cells(1×10^6 cells) into the right footpad of C57BL/6Cr mice Mowed by surgical amputation of the primary tumor 2 weeks after tumor inoculation.为了阐明PKC412抗侵入性作用的机制，我们测量了细胞运动，基质金属蛋白酶（MMP）活性从细胞分泌的活性以及整合蛋白β1蛋白在细胞中的表达。结果，PKC412以剂量依赖性方式降低了B16-BL6细胞的整联蛋白β1蛋白的表达。但是，PKC412无法抑制黑色素瘤细胞的细胞运动和MMP活性。这些结果表明，PKC412通过衰减整合蛋白β1表达抑制黑色素瘤细胞转移性途径的侵袭过程。最后，我们使用特定的PKC Delta抑制剂Rottlerin检查了与转移相关的PKC亚型。由于罗特林无法使用B16-BL6细胞阻断小鼠中的血肿肺转移，因此常规的PKC同工型被认为可以诱导肿瘤转移，而三角洲同工型却没有。