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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Identification of cancer/testis-antigen genes by massively parallel signature sequencing
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Identification of cancer/testis-antigen genes by massively parallel signature sequencing

机译：通过大规模平行签名测序鉴定癌症/睾丸抗原基因

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摘要

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-kb region 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.
机译：大规模并行签名测序（MPSS）生成数百万个短序列标签，对应于单个RNA制备物中的转录本。大多数MPSS标签可以明确分配给基因，从而生成起源组织的全面表达谱。通过比较来自32个正常人体组织的MPSS数据，我们确定了1,056个在睾丸中主要表达的基因。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，将这些基因中的202个确定为编码癌症/睾丸（CT）抗原的候选基因。在这些基因中，通过RT-PCR评估了166个含内含子的基因在正常组织中的表达，并进一步评估了以睾丸为主的表达的基因在21个癌细胞系中的表达。因此，鉴定出20个CT或类似CT的基因，其中几个在五个或多个所检查的癌细胞系中表现出表达。这些基因之一是我们称为CT45的CT基因家族的成员。 CT45家族包含六个高度相似（> 98％的cDNA一致性）的基因，它们在Xq26.3的125kb区域内串联聚集。发现CT45在癌细胞系和肺癌标本中均频繁表达。因此，MPSS分析已大大扩展了我们对CT抗原的认识，导致发现了独特的X连锁CT抗原基因家族。

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