【摘要】 背景与目的:NGAL(neutrophilgelatinase-associatedlipocalin)基因是lipocalin家族的一个新成员。以往我们曾研究发现NGAL基因在TPA诱导永生化食管上皮细胞SHEE向食管癌细胞SHEEC恶性转化的过程中显著过表达,但表达调控机制不清楚。本研究拟从SHEEC细胞中克隆NGAL基因的5’-UTR和3’-UTR(untranslatedregion)并进一步明确其结构特征。方法:采用RACE方法从SHEEC细胞中克隆NGAL基因的5’-UTR和3’-UTR;在测序基础上进行BLAST分析鉴定。结果:结合以往的实验结果,从SHEEC细胞中克隆了NGAL基因的69bp5’-UTR和147bp3’-UTR,而且序列分析未发现突变。结论:SHEEC细胞中的NGAL基因具备完整的5’-UTR和3’-UTR结构。更多还原

【Abstract】 BACKGROUND & OBJECTIVE:Neutrophil gelatinase associated lipocalin (NGAL) was a novel member of the lipocalin family. The authors previously found that NGAL was overexpressed in the progress of malignant transformation from human immortalized esophageal epithelial cell line SHEE to esophageal carcinoma cell line SHEEC. However, the regulation mechanism of NGAL overexpression was not known. The objective of this study was to clone 5′ untranslated region(5′ UTR) and 3′untranslated region (3′ UTR) of NGAL in SHEEC and to analyze their structural characters. METHODS:5′ UTR and 3′ UTR of NGAL were cloned from SHEEC using rapid amplification of cDNA ends(RACE). After sequencing the alignment of their nucleotides was analyzed by BLAST database of NCBI and the potential cis acting elements in the 3′ UTR were identified by computer analysis. RESULTS:The authors cloned and sequenced 69 bp 5′ UTR and 147 bp 3′ UTR of NGAL gene on the basis of the previous works and did not find any base pair mutation. CONCLUSION:NGAL gene from SHEEC had the entire 5′ UTR and 3′ UTR.更多还原