Efficient identification of a novel cancer/testis antigen for immunotherapy using three-step microarray analysis
Efficient identification of a novel cancer/testis antigen for immunotherapy using three-step microarray analysis
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DOI：
10.1158/0008-5472.can-07-0964
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发表时间：
2008-02-15
期刊：
影响因子：
11.2
通讯作者：
Mori, Masaki
中科院分区：
文献类型：
--
作者：
Yokoe, Takeshi;Tanaka, Fumiaki;Mori, Masaki
先进的分子生物学技术为癌症的诊断和治疗提供了强有力的工具。我们采用一种新的方法来鉴定一种在大肠癌中高表达的新的癌/睾丸（cancer/testis，CT）抗原：（a）将激光显微切割和cDNA微阵列技术相结合来分析大肠癌细胞的基因表达谱：（B）用cDNA微阵列技术分析在睾丸中过表达和在正常结肠上皮中低表达的基因;以及（c）将结直肠癌细胞的基因表达谱与正常睾丸的基因表达谱进行比较。使用这种方法，我们选择了38个候选的CT抗原。在这些基因中，我们确定了一个新的CT抗原，丝氨酸/苏氨酸激酶31（STK 31），这是以前报道的基因在精原细胞中表达。RT-PCR分析显示，STK 31基因在胃癌组织中的表达水平显著高于正常组织（P < 0.0001）。STK 31基因不仅在结直肠癌中频繁表达，而且在胃癌和食管癌中也频繁表达。此外，STK 31肽能够引发特异性CTL，并且诱导的CTL裂解肽负载或内源性表达STK 31的靶细胞。这些结果表明，本研究中的新方法有助于鉴定CT抗原，STK 31可能是针对胃肠道癌的癌症免疫治疗的候选者。
Advanced technology in molecular biology has provided us powerful tools for the diagnosis and treatment for cancer. We herein adopted a new methodology to identify a novel cancer/testis (CT) antigen with high frequency of expression in colorectal cancer as follows: (a) combining laser microdissection and cDNA microarray was used to analyze the gene expression profile of colorectal cancer cells; (b) genes over-expressed in testis and underexpressed in normal colon epithelium were analyzed using cDNA microarray; and (c) the gene expression profile of colorectal cancer cells was compared with that of normal testis. Using this methodology, we selected 38 candidates for CT antigen. Among these genes, we identified a novel CT antigen, serine/threonine kinase 31 (STK31), which was previously reported as a gene expressed in spermatogonia. Reverse transcription-PCR analysis showed that STK31 gene expression levels in cancer samples were significantly higher (P < 0.0001) than those in normal samples. The STK31 gene was frequently expressed not only in colorectal cancer but also in gastric and esophageal cancer. Moreover, STK31 peptide was able to elicit specific CTLs and induced CTLs lysed either peptide-loading or endogenously STK31-expressing target cells. These results showed that the new methodology in this study facilitated identification of CT antigens and that STK31 may be a candidate for cancer immunotherapy against gastrointestinal cancer.