Isolation of highly metastatic clone from human oral cancer cell line transfected active form rho gene and Identification of metastasis relation molecule.

从转染活性rho基因的人口腔癌细胞系中分离高转移克隆并鉴定转移相关分子。

• 批准号: 10671875
• 负责人: UMEDA Masahiro
• 金额: $ 2.05万
• 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: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671875/
• 关键词: oral squamous cell carcinoma; oncogene; lymph node metastasis; highly metastatic clone; rho

An activated ras of fos gene has been known to guide metastatic potential expression. Rho, which is one of the ras-like low molecular weight G proteins, may cause an enhancement of cell motility that is essentiality for invasion or metastasis of cancer by decreasing cellular adhesion abilities. The function and action mechanism of rho has been noted recently. Akedo et al. reported that an activated human rho increased invasion ability of rat ascites hepatoma cells into mesothelial cell monolayer in an in vitro experiment.In the other hand, there were few studies of cancer metastasis using oral cancer cell lines. We have searched tumorigenicity by transplanting humans oral cancer cell line in the back of nude mouse, but lymphnode metastasis was not recognized. Highly metastasizing experimental model of oral cancer is necessary for studies of metastasis of oral cancer. So, we transfected plasmid pcDSR αrhoval^<14> that was active form of rho gene into NOS-1, the oral cancer cell line established form the primary squamous cell carcinoma of the lower gingiva, in order to obtain stable transfectants. Further, we examined whether highly metastasizing tumor cell line to the lymph node could be established from these cells.Consequently, stable transfectants which expressed active human rhoA gene were obtained by cotransfecting pcDSR αrhoVal^<14> and drug resistance gene pSV2neo. These clones promoted cell motility 2 times larger than that of the control by a phagokinetic track assay. They were transplantable orthotopically in the tongue of nude mouse, but lymph node metastasis was not observed.In this study, the cell motility of the clones that expressed active form rhoA gene showed an enhancement, but they did not give the lymphogenous metastatic potential.

FOS基因激活的ras基因已被认为可以指导转移潜能的表达。Rho是一种ras样低分子量G蛋白，可能通过降低细胞黏附能力而导致细胞运动性增强，从而对肿瘤的侵袭或转移起重要作用。近年来，Rho的功能和作用机制受到了人们的关注。Akedo等人。报道了一种活化人Rho在体外实验中增加了大鼠腹水肝癌细胞向间皮细胞单层侵袭的能力。另一方面，利用口腔癌细胞系进行肿瘤转移的研究很少。我们用裸鼠背部移植人口腔癌细胞系的方法研究了其致瘤性，但未发现有淋巴结转移。口腔癌高转移实验模型是研究口腔癌转移的必要模型。为此，我们将RHO基因活性形式的真核表达载体pc DSR-α导入口腔癌细胞株NOS1中，以获得稳定的转染体。并将其与耐药基因pSV2neo共转染，获得了稳定表达人RhoA活性基因的细胞株α。通过吞噬动力学跟踪试验，这些克隆对细胞运动的促进作用是对照的2倍。在本研究中，表达活化型RhoA基因的克隆的细胞运动能力有所增强，但并未表现出淋巴转移潜能。