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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。
Objective : To clone target genes regulated by ATRA during differentiation of esophageal squamous carcinoma EC8712 cells. METHODS: AP PCR was used to clone ATRA-regulated differentially expressed gene fragments from esophageal squamous cell carcinoma EC8712, sequenced and homologously analyzed, and verified by Northern hybridization. RESULTS: A gene fragment that was down-regulated during the differentiation of esophageal cancer cells was isolated. The expression of this gene was significantly inhibited from the beginning of ATRA treatment, and this inhibition continued throughout the induction process. This fragment was cloned in the pGEM T vector and sequenced. Homologous analysis results showed that the gene fragment was highly homologous to the human N-ethyl maleimide sensitive factor (NSF) gene (IDENTITY: 99%) and was a human NSF gene fragment. The NSF gene plays an important role in eukaryotic cell vesicle transport system. Conclusion: The NSF gene involved in vesicle transport plays an important role in the differentiation of esophageal cancer cells. The inhibition of NSF expression is the molecular mechanism of the decrease in transport activity of vesicle transport system.