Study of the role of protein kinase C-phosphoipase D signaling pathway in pigment cells.

蛋白激酶C-磷酸酶D信号通路在色素细胞中的作用研究。

• 批准号: 13670886
• 负责人: OKA Masahiro
• 金额: $ 2.3万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670886/
• 关键词: malignant melanoma; protein kinase C; phosphorylation; protein-protein interaction; invasion; 色素細胞; Cキナーゼ; ホスホリパーゼD

It is well known that phospholipase D (PLD) plays a crucial role in the signal transduction of many types of cells, and is activated by protein kinase C α (PKCα) when cells are stimulated. To elucidate the role of PLD in melanoma, the expression of PLD1 and PKCα in primary and metastatic lesions of acral lentiginous melanoma (ALM) and superficial spreading melanoma (SSM) was investigated using immunohistological techniques. In addition, the mechanism of regulation of PLD1 by PKCα was examined in a human melanoma cell line HM3KO using an adenovirus-mediated gene transfer technique. Both PLD1 and DKCα were strongly expressed in primary and metastatic lesions of SSM. Conversely, in ALM lesions, the expression of these two proteins increased dramatically with tumor progression ; the expression of both PLD1 and PKCα was almost negative in the radial growth phase of primary ALM lesions, and increased synchronously in a progression-related manner in advanced ALM esions, including vertical gro … More wth phase and metastatic lesions. Immunoprecipitation study showed that PLD1 and PKCα are associated physiologically in resting melanoma cells. Further immunoprecipitation study using HM3KO cells after adenovirus-mediated simultaneous overexpression of PLD1 and PKCα, or PLD1 and the kinase-negative mutant of PKCα revealed that both PKCα and the kinase-negative mutant of PKCα are associated with PLD1 in melanoma cells in the absence of an extemal signal. Overexpression of PKCα or the kinase-negative mutant of PKCα in melanoma cells by the adenovirus vectors resulted in the enhancement of basal PLD activity in a viral dose-dependent manner. Furthermore, enhanced basal PLD activity increased the in vitro invasive potential of HM3KO cells. These results suggest that upregulation of PLD1 and PKCα plays a rote in the progression of ALM from the radial growth phase to the vertical growth phase. The present results also suggest that PKCα associates with PLD1 and enhances basal PLD activity in a protein phosphorylation-independent manner in melanoma cells, which contributes to the cell's high invasive potential. Less

众所周知，磷脂酶D（PLD）在许多类型的细胞的信号转导中起着至关重要的作用，当刺激细胞时，蛋白激酶Cα（PKCα）被蛋白激酶Cα（PKCα）激活。为了阐明PLD在黑色素瘤中的作用，使用免疫组织学技术研究了PLD1和PKCα在丙二矿物质黑色素瘤（ALM）（ALM）的原发和转移性病变中的表达。此外，使用腺病毒介导的基因转移技术在人黑色素瘤细胞系HM3KO中检查了PKCα调节PLD1的机理。 PLD1和DKCα都在SSM的原发性和转移性病变中强烈表达。相反，在ALM病变中，这两种蛋白质的表达随肿瘤进展而显着增加。在原发性ALM病变的径向生长阶段，PLD1和PKCα的表达几乎为阴性，并且在晚期ALM Essions中以与进展相关的方式同步增加，包括垂直Gro…更多的WTH相和转移性病变。免疫沉淀研究表明，PLD1和PKCα与静息黑色素瘤细胞物理相关。腺病毒介导的PLD1和PKCα或PLD1的同时过表达后，使用HM3KO细胞进行进一步的免疫沉淀研究，PKCα的激酶阴性突变体均表明，PLD1的PKCα和PKCα的激酶阴性突变剂都与PLD1在墨拉哥细胞中的PLD1相关。腺病毒载体中PKCα或PKCα的激酶阴性突变体的过表达导致以病毒剂量依赖性方式增强碱性PLD活性。此外，增强的碱性PLD活性增加了HM3KO细胞的体外浸润潜力。这些结果表明，PLD1和PKCα的上调在ALM从径向生长阶段到垂直生长阶段的腐烂作用。目前的结果还表明，PKCα与PLD1相关并增强了黑色素瘤细胞中蛋白质磷酸化非依赖性方式的碱性PLD活性，这有助于细胞的高浸润潜力。较少的

项目成果

Oka, M.: "Dual regulation of phospholipase D1 by protein kinase Cα in vivo"Biochem. Biophys. Res. Commun.. 294・5. 1109-1113 (2001)

Oka, M.：“体内蛋白激酶 Cα 的双重调节”Biochem. Commun. 1109-1113 (2001)。

Oka, M.: "Dual regulation of phospholipase D1 by protein kinase Cα in vivo"Biochem.Biophys.Res.Commun.. 294. 1109-1113 (2002)

Oka, M.：“体内蛋白激酶 Cα 对磷脂酶 D1 的双重调节”Biochem.Biophys.Res.Commun. 294. 1109-1113 (2002)

Nagai, H.: "Gene transfer of secreted-type modified interleukin-18 gene to B16F10 melanoma cells suppresses in vivo tumor growth through inhibition of tumor vessel formation"J. Invest. Dermatol.. 119・3. 541-548 (2002)

Nagai, H.：“将分泌型修饰的白细胞介素 18 基因转移至 B16F10 黑色素瘤细胞，通过抑制肿瘤血管形成来抑制体内肿瘤生长”J. Invest. 119・3 (2002)。

Mizuho Fukunaga et al.: "UV-Induced Tyrosine Phosphorylation of PKCS and Promstion of Apoptosis in the HaCaT Cell Line"Biochemical And Biophysical Research Communications. 289・2. 573-579 (2001)

Mizuho Fukunaga 等人：“HaCaT 细胞系中 PKCS 的紫外线诱导酪氨酸磷酸化和细胞凋亡的促进”生物化学和生物物理研究通讯 289・2（2001）。

Nagai, H.: "Gene transfer of secreted-type modified interleukin-18 gene to B16F10 melanoma cells suppresses in vivo tumor growth through inhibition of tumor vessel formation"J.Invest Dermatol.. 119. 541-548 (2002)

Nagai, H.：“将分泌型修饰的白细胞介素 18 基因转移至 B16F10 黑色素瘤细胞，通过抑制肿瘤血管形成来抑制体内肿瘤生长”J.Invest Dermatol.. 119. 541-548 (2002)