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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.

我们研究了蛋白激酶C（PKC）抑制剂抗肿瘤转移的机制。PKC 412（4 '-N-苯甲酰基星形孢菌素）是一种常规PKC（α、β和γ）抑制剂，当与细胞预孵育1小时时，可降低小鼠恶性黑色素瘤（B16-BL 6）细胞在小鼠体内形成肺集落的能力，并减少体外细胞外基质的侵袭。此外，PKC 412（200 mg/kg/天，持续4周，p.o.）在自发转移性小鼠模型中显著延长了生存时间，该模型通过将B16-BL 6细胞（1×10^6个细胞）皮下接种到C57 BL/6Cr小鼠的右足垫中产生，在肿瘤接种后2周通过手术切除原发性肿瘤进行切除。为了阐明PKC 412抗侵袭作用的机制，我们检测了细胞运动性、细胞分泌的基质金属蛋白酶（MMP）活性和细胞中整合素β 1蛋白的表达。结果表明，PKC 412可剂量依赖性地降低B16-BL 6细胞整合素β 1蛋白的表达。但PKC 412不能抑制黑色素瘤细胞的运动和MMP活性。这些结果表明，PKC 412抑制黑色素瘤细胞的转移途径的整合素β 1的表达衰减的侵袭过程。最后，我们研究了转移相关的PKC亚型使用rottlerin，一个特定的PKC δ抑制剂。由于rottlerin不能阻断使用B16-BL 6细胞的小鼠中的血液源性肺转移，因此认为常规PKC亚型诱导肿瘤转移，而δ亚型不诱导肿瘤转移。