Modulation of Cyclin D1 Expression by Integrin Signals in Oral Squamous Carcinoma Cells

口腔鳞状细胞癌细胞中整合素信号对细胞周期蛋白 D1 表达的调节

• 批准号: 15592000
• 负责人: KAWANO Kenji
• 金额: $ 2.11万
• 依托单位: Oita University (2004-2005)大分医科大学 (2003)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15592000/
• 关键词: Oral sqaumous cell carcinoma; Cyclin D1; GSK-3β; integrin; Extracellular matrix; PI3K-Akt pathway; Ras-MAPK pathway; Akt; Erk; PI3K; セリン残基リン酸化

Cyclin D1 overexpression is frequently detected in oral squamous cell carcinomas (OSCC) and it is closely associated with the survival of OSCC patients. We examined cyclin D1 gene amplification with a panel of OSCC cell lines by the differential PCR technique. Any OSCC cell lines examined, however, did not show gene amplification of cyclin D1. Then, the expression of phosphorylated GSK-3β was examined by Western blotting, and was compared with cyclin D1 expression in OSCC cells. Interestingly, the expression of phosphorylated GSK-3β was reversely correlated with cyclin D1 expression, suggesting that lack of phosphorylated GSK-3β causes the intracellular accumulation of cyclin D1 in OSCC cells.Next, we tested cyclin D1 expression in various culture conditions, such as culturing with 10% FBS-containing medium or serum-free medium, as monolayer culture cells or multicellular aggregates (MCA), and on specific extracellular matrix proteins-coated surfaces or non-coated surface, and we found no differences in cyclin D1 level under these culture conditions. Furthermore, localizations of cyclin D1, GSK-3, Akt and Erk were investigated in monolayer cells and MCAs by immunofluorescent stainings. Results showed diffuse positive reactions without any differences between these proteins and between both the culture conditions.In this project, we could not present definite evidence of involvement of integrin signals in cyclin D1 expression in OSCC cell lines. Further studies are required to clarify the mechanism of modulation of GSK-3β functionality in OSCC.

口腔鳞状细胞癌（oral squamous cell carcinoma，OSCC）中细胞周期蛋白D1（Cyclin D1）的过度表达与患者的生存密切相关。我们用差异PCR技术检测了一组口腔鳞癌细胞系的细胞周期蛋白D1基因扩增。然而，任何口腔鳞状细胞癌细胞系检查，并没有显示细胞周期蛋白D1的基因扩增。Western blotting检测磷酸化GSK-3β的表达，并与cyclin D1的表达进行比较。有趣的是，磷酸化GSK-3β的表达与细胞周期蛋白D1的表达呈负相关，这表明磷酸化GSK-3β的缺乏导致细胞周期蛋白D1在OSCC细胞中的细胞内积累。接下来，我们测试了细胞周期蛋白D1在各种培养条件下的表达，例如在含10%FBS的培养基或无血清培养基中培养，作为单层培养细胞或多细胞聚集体（MCA），在特定的细胞外基质蛋白包被的表面或非包被的表面上，我们发现在这些培养条件下细胞周期蛋白D1的水平没有差异。此外，通过免疫荧光染色研究了细胞周期蛋白D1、GSK-3、Akt和Erk在单层细胞和MCA中的定位。结果显示，这些蛋白之间和两种培养条件之间没有任何差异的弥漫阳性反应。在本项目中，我们不能提出明确的证据，整合素信号参与细胞周期蛋白D1在口腔鳞癌细胞系的表达。GSK-3β在口腔鳞癌中的作用机制有待进一步研究。

项目成果

Predictive value of laminin-5 and MT1-MMP expression for cervical lymphnode metastasis in T1 and T2 squamous cell carcinomas of the tongue and floor of the mouth

Laminin-5和MT1-MMP表达对T1、T2期舌、口底鳞状细胞癌颈部淋巴结转移的预测价值

• 发表时间: 2006
• 期刊: Head and Neck 28(In press)
• 作者: Kenji Kawano;Shigetaka Yanagisawa
• 通讯作者: Shigetaka Yanagisawa

Roles of N-cadherin in invasion and metastasis of E-cadherin-negative oral squamous carcinoma cell lines

N-cadherin在E-cadherin阴性口腔鳞癌细胞系侵袭和转移中的作用

• 发表时间: 2005
• 期刊: Oral Oncology ; Proceedings of 10th ICOOC 10
• 作者: Yamazaki M;Ohshiro K;Cheng J;Ida H;Kawamura H;Saku T;Morisaki I;Kenji Kawano et al.
• 通讯作者: Kenji Kawano et al.

Kenji Kawano et al.: "Predictive value of immunohistochemical thymidylate synthase expression for histological response to Tegafur/Uracil (UFT) in oral squamous cell carcinoma"International Journal of Oral & Maxillofacial Surgery. 32・6. 633-637 (2003)

Kenji Kawano 等人：“免疫组织化学胸苷酸合酶表达对口腔鳞状细胞癌替加氟/尿嘧啶 (UFT) 组织学反应的预测价值”国际口腔颌面外科杂志 32・6 (2003)。

Intratumoral expression of thymidylate synthase is an independent Predictor of prognosis in patients with squamous cell carcinoma of the tongue : results from a retrospective study

胸苷酸合酶的肿瘤内表达是舌鳞状细胞癌患者预后的独立预测因子：回顾性研究的结果

• 发表时间: 2006
• 期刊: International Journal of Oral & Maxillofacial Surgery 35
• 作者: Kenji Kawano et al.