【摘要】 目的 :克隆食管鳞癌细胞EC8712分化过程中受ATRA调控的靶基因。方法 :采用AP PCR技术从食管鳞癌细胞EC8712中克隆受ATRA调控的差异表达基因片段 ,并测序及作同源分析 ,采用Northern杂交进行验证。结果 :分离到在食管癌细胞分化过程中表达下调的一基因片段。该基因从ATRA处理一天开始表达被明显抑制 ,这种抑制持续整个诱导过程。该片段被克隆于 pGEM T载体并测序。同源分析结果表明 ,该基因片段与人N 乙基马来酰亚胺敏感因子 (NSF)基因高度同源 (IDENTITY :99% ) ,为人NSF基因片段。NSF基因在真核细胞囊泡转运系统中发挥着重要作用。结论 :参与囊泡转运的NSF基因在食管癌细胞分化过程中发挥着重要作用 ,NSF表达的抑制是囊泡转运系统转运活性下降的分子机制。更多还原

【Abstract】 Objective:To clone ATRA regulated gene in esophageal squamous cancer cell line EC8712 Methods:By using AP PCR technique, differentially expressed gene fragments (ESTs) were isolated from ATRA treated EC8712 Cloning, sequencing and homology analysis were carried out to characterize an isolated differentitally expressed gene fragment Results:The differentially expressed gene fragment was isolated, cloned and sequenced, the gene expression was inhibited during whole ATRA treatment period, from first day to 9th day Homology analysis revealed that the fragment derives from human N ethylmalimide sensitive factor (NSF) gene, which is involed in vesicle transportation, expresses in various tissues, and plays important roles in differentiation of cells Conclusion:The decrease in activity of cellular vesicle transportation plays important roles in cellular differentiation process Down regulation of NSF might contribute to decreased activity of vesicle transportation and differentiation of human cancer cell更多还原