Identification of cancer/testis-antigen genes by massively parallel signature sequencing
Identification of cancer/testis-antigen genes by massively parallel signature sequencing
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DOI：
10.1073/pnas.0502583102
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发表时间：
2005-05-31
影响因子：
11.1
通讯作者：
Simpson, AJG
中科院分区：
文献类型：
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作者：
Chen, YT;Scanlan, MJ;Simpson, AJG
大规模平行签名测序（MPSS）产生数百万个短序列标签，对应于单个RNA制备的转录本。大多数MPSS标签可以明确地分配给基因，从而产生起源组织的全面表达谱。通过对32个正常人体组织的MPSS数据的比较，我们确定了1056个主要在睾丸中表达的基因。通过使用来自癌细胞系的MPSS标签和来自多种肿瘤的EST数据进行进一步评估，鉴定出202个这些基因作为编码癌/睾丸（CT）抗原的候选基因。在这些基因中，通过RT-PCR检测了166个含内含子基因在正常组织中的表达情况，并进一步检测了在21个癌细胞系中睾丸显性表达的基因的表达情况。因此，鉴定了20个CT或CT样基因，其中几个在检查的五种或更多癌细胞系中表现出表达。其中一个基因是CT基因家族的成员，我们称之为CT45。CT45家族包括6个高度相似（>； 98% cDNA同源性）的基因，它们串联聚集在Xq26.3上的一个125 kb区域内。发现CT45在癌细胞系和肺癌标本中都频繁表达。因此，MPSS分析大大扩展了我们对CT抗原的认识，从而发现了一个独特的x连锁CT抗原基因家族。
Massively parallel signature sequencing (MPSS) generates millions of short sequence tags corresponding to transcripts from a single RNA preparation. Most MPSS tags can be unambiguously assigned to genes, thereby generating a comprehensive expression profile of the tissue of origin. From the comparison of MPSS data from 32 normal human tissues, we identified 1,056 genes that are predominantly expressed in the testis. Further evaluation by using MPSS tags from cancer cell lines and EST data from a wide variety of tumors identified 202 of these genes as candidates for encoding cancer/testis (CT) antigens. Of these genes, the expression in normal tissues was assessed by RT-PCR in a subset of 166 intron-containing genes, and those with confirmed testis-predominant expression were further evaluated for their expression in 21 cancer cell lines. Thus, 20 CT or CT-like genes were identified,with several exhibiting expression in five or more of the cancer cell lines examined. One of these genes is a member of a CT gene family that we designated as CT45. The CT45 family comprises six highly similar (> 98% cDNA identity) genes that are clustered in tandem within a 125-kbregion on Xq26.3. CT45 was found to be frequently expressed in both cancer cell lines and lung cancer specimens. Thus, MPSS analysis has resulted in a significant extension of our knowledge of CT antigens, leading to the discovery of a distinctive X-linked CT-antigen gene family.