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[利用抑制性消减杂交技术克隆的肺鳞状细胞癌相关基因]

[Lung squamous cell carcinoma-related genes cloned by using suppression subtractive hybridization].

作者信息

Liu Jun, Shen Chen-yang, Zhao Hui, Wang Dan-lei, Zhang Guo-liang, Wang Jun

机构信息

Department of Thoracic Surgery, People's Hospital, Peking University, Beijing 100044, China.

出版信息

Zhonghua Wai Ke Za Zhi. 2003 May;41(5):368-71.

Abstract

OBJECTIVE

To identify genes differentially expressed in human lung squamous cell carcinoma (LSCC).

METHODS

A subtracted cDNA library of human LSCC constructed by using suppression subtracted hybridization (SSH) method was screened. Clones representing mRNAs that were truly differentially expressed in LSCC but not in its adjacent non-cancerous tissues were identified by semi-quantitative RT-PCR in 12 patients with LSCC. Partial novel genes were detected by Northern blot.

RESULTS

Ten differentially expressed gene cDNA fragments of LSSC were obtained by SSH. Among them six were known genes; two sequences were identified but their functions were unknown (hypothetical protein); two were novel (GenBank accession numbers AF363068 and AY032661, respectively). The results from semiquantitative RT-PCR showed that the transcription expression level of PPP1CB, calumenin, S100A2, HSNOV1, OCIA and AY032661 was down-regulated in some LSCC cases, while the transcription of HSP90, ferritin, gp96 and AF363068 was up-regulated in others.

CONCLUSIONS

Six known genes identified by SSH technique have been implicated in the pathogenesis of lung carcinogenesis, or they are involved in immunological defense mechanism in human body. Two hypothetical proteins probably also play an important role in the pathogenesis of lung cancer. The function of the two novel genes in lung carcinogenesis are under investigation.

摘要

目的

鉴定在人肺鳞状细胞癌(LSCC)中差异表达的基因。

方法

筛选利用抑制性消减杂交(SSH)方法构建的人LSCC消减cDNA文库。通过半定量RT-PCR在12例LSCC患者中鉴定出代表在LSCC中真正差异表达但在其相邻非癌组织中不表达的mRNA的克隆。通过Northern印迹检测部分新基因。

结果

通过SSH获得了10个LSCC差异表达基因的cDNA片段。其中6个是已知基因;2个序列已鉴定但功能未知(假设蛋白);2个是新基因(GenBank登录号分别为AF363068和AY032661)。半定量RT-PCR结果显示,在一些LSCC病例中PPP1CB、钙网蛋白、S100A2、HSNOV1、OCIA和AY032661的转录表达水平下调,而在其他病例中HSP90、铁蛋白、gp96和AF363068的转录上调。

结论

通过SSH技术鉴定的6个已知基因与肺癌发生的发病机制有关,或参与人体免疫防御机制。两种假设蛋白可能在肺癌发病机制中也起重要作用。这两个新基因在肺癌发生中的功能正在研究中。

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