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用于检测头颈癌中eIF4E基因扩增的竞争性聚合酶链反应

Competitive PCR to detect eIF4E gene amplification in head and neck cancer.

作者信息

Sorrells D L, Ghali G E, Meschonat C, DeFatta R J, Black D, Liu L, De Benedetti A, Nathan C A, Li B D

机构信息

Department of Surgery, Louisiana State University Medical Center, Shreveport 71130, USA.

出版信息

Head Neck. 1999 Jan;21(1):60-5. doi: 10.1002/(sici)1097-0347(199901)21:1<60::aid-hed8>3.0.co;2-j.

Abstract

BACKGROUND

The protein eukaryotic initiation factor 4E (elF4E) binds to messenger ribonucleic acid (mRNA) as the initial step in protein synthesis. Overexpression of elF4E results in upregulation of specific proteins essential to cell growth and division. Overexpression of elF4E has been found in head and neck squamous cell carcinoma (HNSCC) and breast carcinoma. This study's purpose is to determine whether elF4E overexpression is present and associated with elF4E gene amplification in HNSCC.

METHODS

Competitive polymerase chain reaction (PCR) was performed on eight HNSCC and seven intraoral benign lesions to determine the copy number of elF4E relative to a reference gene, gastrin. Western blots were performed to quantify elF4E protein expression.

RESULTS

All eight HNSCC specimens demonstrated significant (p < .005) overexpression of elF4E protein (14.1+/-10.4) and elF4E gene amplification (4.5+/-1.2). Benign tissue did not exhibit elF4E protein overexpression or gene amplification.

CONCLUSIONS

Overexpression and associated gene amplification of elF4E were present in HNSCC but not in benign tissue. Gene amplification of elF4E may be an important mechanism for elF4E overexpression.

摘要

背景

真核生物起始因子4E(elF4E)蛋白与信使核糖核酸(mRNA)结合,是蛋白质合成的起始步骤。elF4E的过表达会导致细胞生长和分裂所必需的特定蛋白质上调。在头颈部鳞状细胞癌(HNSCC)和乳腺癌中已发现elF4E过表达。本研究的目的是确定HNSCC中是否存在elF4E过表达及其与elF4E基因扩增的关系。

方法

对8例HNSCC和7例口腔良性病变进行竞争性聚合酶链反应(PCR),以确定相对于参考基因胃泌素的elF4E拷贝数。进行蛋白质免疫印迹以定量elF4E蛋白表达。

结果

所有8例HNSCC标本均显示elF4E蛋白显著(p <.005)过表达(14.1±10.4)和elF4E基因扩增(4.5±1.2)。良性组织未表现出elF4E蛋白过表达或基因扩增。

结论

HNSCC中存在elF4E过表达及相关基因扩增,而良性组织中不存在。elF4E基因扩增可能是elF4E过表达的重要机制。

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