Identification of a novel transcript up-regulated in a clinically aggressive prostate carcinoma
Identification of a novel transcript up-regulated in a clinically aggressive prostate carcinoma
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DOI：
10.1016/s0090-4295(97)00194-5
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发表时间：
1997-08-01
期刊：
影响因子：
2.1
通讯作者：
EmmertBuck, MR
中科院分区：
文献类型：
--
作者：
Chuaqui, RF;Englert, CR;EmmertBuck, MR
目标.采用组织显微切割和靶向差异显示技术，筛选前列腺癌患者肿瘤细胞中差异表达的基因。从根治性直肠癌切除术标本的冷冻组织切片的显微切割的正常上皮和侵袭性肿瘤的纯群体中回收RNA。结果：用随机引物和锌指PCR引物进行逆转录聚合酶链反应（PCR），在肿瘤组织中特异性扩增出一个130 bp的产物。DNA序列分析显示它是来自表达序列标签数据库（GenBank登录号R 00504）的克隆。正常上皮和相应的浸润性肿瘤的显微解剖，随后进行了测试面板上的10个前列腺癌标本。使用β-肌动蛋白作为内部对照，比较正常上皮和浸润性癌中的R 00504水平，显示转录本在10种癌中的5种中显著过表达。北方杂交结果表明R 00504是一个2.6-脱氢酶基因。一个肚脐转录上调的侵袭性前列腺癌，确定使用简并锌指引物在显微组织样本。本研究所用的方法可能有助于已知基因的定量比较和新凝胶的鉴定。在显微解剖的人体组织样本中。(C)1997年，Elsevier Science Inc. All rights reserved.
Objectives. To identify differentially expressed genes in tumor cells of patients with prostate cancer by means of tissue microdissection and targeted differential display.Methods. RNA was recovered from pure populations of microdissected normal epithelium and invasive tumor from frozen tissue sections of a radical prostatectomy specimen. Reverse transcriptian-polymerase chain reaction (PCR) using arbitrary and zinc finger PCR primers was performed.Results, A 130-base pair product was identified that appeared selectively in the tumor sample. DNA sequence analysis revealed it to be a clone from the expressed sequence tag database (GenBank accession R00504). Microdissection of normal epithelium and the corresponding invasive tumor was subsequently performed on a test panel of 10 prostate carcinoma specimens. Comparison of R00504 levels in normal epithelium and invasive carcinoma, using beta-actin as an internal control, showed the transcript to be substantially overexpressed in 5 of 10 carcinomas. Northern blotting revealed R00504 to be a 2.6-kilobase gene.Conclusions. A navel transcript up-regulated in an aggressive prostate carcinoma was identified using degenerate zinc finger primers in microdissected tissue samples. The approach used in this study may be helpful in quantitative comparison of known genes and identification of novel gel?es in microdissected human tissue samples. (C) 1997, Elsevier Science Inc. All rights reserved.