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

1：在鳞癌细胞中以Sox-2作为干细胞因子的致癌转激活靶基因的搜索和鉴定：我们使用affmetrix微阵列芯片对293细胞中表达Sox-2或不表达Sox-2的总RNA进行比较，获得图谱。在该图谱中，我们发现两个候选癌基因被Sox-2表达转激活，两个肿瘤抑制基因被其表达转抑制。II：检测调节Sox-2表达的基因：我们克隆了一个约500 bp的Sox-2基因启动子区域片段。我们将该片段亚克隆到pGL3载体中以分析启动子。因此，我们将能够通过分析检测转录激活因子或Sox-2基因表达的抑制因子。三、Sox-2基因作为鳞状细胞癌干细胞因子的临床应用：我们通过围手术期外周血中Sox-2 mRNA的定量检测，研究其作为鳞状细胞癌肿瘤标志物的可能性。8例食管癌患者中有5例术后14天外周血中已检测不到Sox-2 mRNA。IV: Sox-2基因的基因扩增：Sox-2基因位于3q26-27，在各种肿瘤组织中均检测到基因组扩增。我们用Southern blot技术检测了60例食管癌中12例的基因组扩增。这一发现表明，Sox-2基因是鳞状细胞癌的致癌基因。

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.