Detection and clinical application of Stem Cell Factor in Squamous cell tissue and cancer.

鳞状细胞组织和癌症中干细胞因子的检测及临床应用。

• 批准号: 17591391
• 负责人: NOMOTO Shuji
• 金额: $ 2.24万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591391/
• 关键词: squamous cell factor; Stem Cell Factor; Sox-2 gene; expression; mRNA; protein; promoter; tumor marker

I : Search and identification of the oncogenic transactivated target genes by Sox-2 as the Stem cell factor in squamous cancer cells : We compared the total RNA with or without Sox-2 expression in 293 cells using Affimetrix micro-array chip to obtain the profile. In the profile, we found two candidate oncogenes transactivated by Sox-2 expression, and two tumor suppressor genes transrepressed by its expression.II : Detecting genes which regulate Sox-2 expression: We cloned an about 500 bp fragment of the promoter region of Sox-2 gene. We sub-cloned the fragment into the pGL3 vector to analyze a promoter. We thus will be able to detect transcription activators or repressors of Sox-2 gene expression by the analysis.III : Clinical application of Sox-2 gene as the Stem cell factor for squamous cell cancer : We studied the possibilities of its usefulness as a tumor marker for squamous cell cancer by quantitative detection of Sox-2 mRNA in perioperative peripheral blood. Sox-2 mRNA which was detectable in the peripheral blood before operation, could no longer be detected at 14 postoperative days in 5 of 8 esophageal cancer patients.IV : Gene amplification of Sox-2 gene: Sox-2 gene locates at 3q26-27, where genome amplification is detected in various cancer tissues. We detected genome amplification in 12 of 60 cases in esophageal cancer by Southern blot technique. The finding suggests that Sox-2 gene is an oncogene for the squamous cell cancer.

SOX-2作为鳞癌细胞干细胞因子的致癌反式激活靶基因的寻找和鉴定：我们利用Affimetrix微阵列芯片比较了293细胞中SOX-2表达前后的总RNA，以获得该基因的表达谱。在这个图谱中，我们发现了两个受SOX-2表达反式激活的候选癌基因，以及两个受其表达反式抑制的肿瘤抑制基因。II：检测调控SOX-2表达的基因：我们克隆了SOX-2基因启动子区域约500bp的片段。我们将该片段亚克隆到pGL3载体中，对启动子进行分析。因此，我们将能够通过分析来检测SOX-2基因表达的转录激活或抑制因子。III：SOX-2基因作为鳞状细胞癌干细胞因子的临床应用：我们通过定量检测围手术期外周血中SOX-2基因的mRNA，研究了其作为鳞癌肿瘤标志物的可能性。8例食道癌患者手术前外周血中检测到SOX-2基因，术后14天有5例患者检测不到SOX-2基因。IV：SOX-2基因扩增：SOX-2基因位于3q26-27，在多种癌组织中均可检测到基因组扩增。应用Southern杂交技术对60例食管癌组织中的12例进行了基因组扩增。这一发现表明SOX-2基因是鳞状细胞癌的癌基因。