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食管癌细胞SHEEC中NGAL基因5'-UTR和3'-UTR的克隆与鉴定 被引量:9

Cloning and Identification of 5-′Untranslated Region (UTR)and 3′-Untranslated Region of Neutrophil Gelatinase-Associated Lipocalin (NGAL) Gene from Esophageal Carcinoma Cell Line SHEEC
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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。 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.
出处 《癌症》 SCIE CAS CSCD 北大核心 2003年第2期143-147,共5页 Chinese Journal of Cancer
基金 国家自然科学基金项目(39900069 30170428) 广东省自然科学基金资助项目(010431) 广东省高校自然科学研究项目(200033) 广东省医学科研基金项目(A2001419) 汕头大学研究与发展基金项目(L0004 L00012)
关键词 食管肿瘤 NGAL基因 基因过表达 转录非翻译区 RACE法 克隆 Esophageal carcinoma Gene overexpression Untranslated Region RACE
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