KIF5B-ALK, a Novel Fusion Oncokinase Identified by an Immunohistochemistry-based Diagnostic System for ALK-positive Lung Cancer
KIF5B-ALK, a Novel Fusion Oncokinase Identified by an Immunohistochemistry-based Diagnostic System for ALK-positive Lung Cancer
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DOI：
10.1158/1078-0432.ccr-08-3248
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发表时间：
2009-05-01
影响因子：
11.5
通讯作者：
Mano, Hiroyuki
中科院分区：
文献类型：
--
作者：
Takeuchi, Kengo;Choi, Young Lim;Mano, Hiroyuki
目的：EML 4-ALK是一种转化融合酪氨酸激酶，在肺癌中发现了多种亚型。然而，EML 4-ALK的免疫组织化学检测已被证明是困难的，这可能是由于EML 4的启动子-增强子区域赋予的低转录活性。EML 4-ALK免疫组化检测的灵敏度应适当增加。实验设计：我们开发了一种嵌入抗体增强聚合物（iAEP）方法，该方法在ALK的一抗和基于葡聚糖聚合物的检测试剂之间加入了一种嵌入抗体。在肺腺癌档案中也发现4个肿瘤ALK阳性（n = 130），在诊断实验室分析的新鲜病例中发现另外4个肿瘤ALK阳性。发现这8例肿瘤包括1例EML 4-ALK变体1、1例变体2、3例变体3和2例先前未鉴定的变体（指定为变体6和7）。反转录-PCR分析显示，剩余的肿瘤具有一种新的融合，其中KIF 5 B的内含子24连接到ALK的内含子19。多重逆转录-PCR分析额外的存档肿瘤标本确定了另一例肺腺癌阳性KIF 5 B-ALK.Conclusions：iAEP方法应证明适合免疫组化筛选肿瘤阳性ALK或ALK融合蛋白之间的病理档案。将基于PCR的检测与iAEP方法相结合，应进一步促进新型ALK融合基因（如KIF 5 B-ALK）的快速鉴定。
Purpose: EML4-ALK is a transforming fusion tyrosine kinase, several isoforms of which have been identified in lung cancer. Immunohistochemical detection of EML4-ALK has proved difficult, however, likely as a result of low transcriptional activity conferred by the promoter-enhancer region of EML4. The sensitivity of EML4-ALK detection by immunohistochemistry should be increased adequately.Experimental Design: We developed an intercalated antibody-enhanced polymer (iAEP) method that incorporates an intercalating antibody between the primary antibody to ALK and the dextran polymer-based detection reagents.Results: Our iAEP method discriminated between tumors positive or negative for EML4-ALK in a test set of specimens. Four tumors were also found to be positive for ALK in an archive of lung adenocarcinoma (n = 130) and another 4 among fresh cases analyzed in a diagnostic laboratory. These 8 tumors were found to include 1 with EML4-ALK variant 1, 1 with variant 2, 3 with variant 3, and 2 with previously unidentified variants (designated variants 6 and 7). Inverse reverse transcription-PCR analysis revealed that the remaining tumor harbored a novel fusion in which intron 24 of KIF5B was ligated to intron 19 of ALK. Multiplex reverse transcription-PCR analysis of additional archival tumor specimens identified another case of lung adenocarcinoma positive for KIF5B-ALK.Conclusions: The iAEP method should prove suitable for immunohistochemical screening of tumors positive for ALK or ALK fusion proteins among pathologic archives. Coupling of PCR-based detection to the iAEP method should further facilitate the rapid identification of novel ALK fusion genes such as KIF5B-ALK.