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利用抑制性消减杂交技术鉴定人肺鳞状细胞癌中差异表达的基因

Identification of differentially expressed genes in human lung squamous cell carcinoma using suppression subtractive hybridization.

作者信息

Sun Wenyue, Zhang Kaitai, Zhang Xinyu, Lei Wendong, Xiao Ting, Ma Jinfang, Guo Suping, Shao Shujuan, Zhang Husheng, Liu Yan, Yuan Jinsong, Hu Zhi, Ma Ying, Feng Xiaoli, Hu Songnian, Zhou Jun, Cheng Shujun, Gao Yanning

机构信息

Department of Etiology and Carcinogenesis, Cancer Institute (Hospital), Peking Union Medical College & Chinese Academy of Medical Sciences, P.O. Box 2258, Beijing 100021, China.

出版信息

Cancer Lett. 2004 Aug 20;212(1):83-93. doi: 10.1016/j.canlet.2004.03.023.

Abstract

Lung cancer is one of the major causes of cancer-related deaths. Over the past decade, much has been known about the molecular changes associated with lung carcinogenesis; however, our understanding to lung tumorigenesis is still incomplete. To identify genes that are differentially expressed in squamous cell carcinoma (SCC) of the lung, we compared the expression profiles between primarily cultured SCC tumor cells and bronchial epithelial cells derived from morphologically normal bronchial epithelium of the same patient. Using suppression subtractive hybridization (SSH), two cDNA libraries containing up- and down-regulated genes in the tumor cells were constructed, named as LCTP and LCBP. The two libraries comprise 258 known genes and 133 unknown genes in total. The known up-regulated genes in the library LCTP represented a variety of functional groups; including metabolism-, cell adhesion and migration-, signal transduction-, and anti-apoptosis-related genes. Using semi-quantitative reverse transcription-polymerase chain reaction, seven genes chosen randomly from the LCTP were analyzed in the tumor tissue paired with its corresponding adjacent normal lung tissue derived from 16 cases of the SCC. Among them, the IQGAP1, RAP1GDS1, PAICS, MLF1, and MARK1 genes showed a consistent expression pattern with that of the SSH analysis. Identification and further characterization of these genes may allow a better understanding of lung carcinogenesis.

摘要

肺癌是癌症相关死亡的主要原因之一。在过去十年中,我们对与肺癌发生相关的分子变化已有很多了解;然而,我们对肺肿瘤发生的认识仍不完整。为了鉴定在肺鳞状细胞癌(SCC)中差异表达的基因,我们比较了原代培养的SCC肿瘤细胞与来自同一患者形态学正常支气管上皮的支气管上皮细胞之间的表达谱。利用抑制性消减杂交(SSH)技术,构建了两个包含肿瘤细胞中上调和下调基因的cDNA文库,分别命名为LCTP和LCBP。这两个文库总共包含258个已知基因和133个未知基因。文库LCTP中已知的上调基因代表了多种功能类别,包括代谢、细胞黏附和迁移、信号转导以及抗凋亡相关基因。使用半定量逆转录-聚合酶链反应,对从16例SCC患者的肿瘤组织及其相应的相邻正常肺组织中随机选取的7个来自LCTP的基因进行了分析。其中,IQGAP1、RAP1GDS1、PAICS、MLF1和MARK1基因的表达模式与SSH分析结果一致。对这些基因的鉴定和进一步表征可能有助于更好地理解肺癌的发生机制。

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