A Prospective PCR-Based Screening for the EML4-ALK Oncogene in Non-Small Cell Lung Cancer
A Prospective PCR-Based Screening for the EML4-ALK Oncogene in Non-Small Cell Lung Cancer
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DOI：
10.1158/1078-0432.ccr-11-2947
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发表时间：
2012-10-15
影响因子：
11.5
通讯作者：
Mano, Hiroyuki
中科院分区：
文献类型：
--
作者：
Soda, Manabu;Isobe, Kazutoshi;Mano, Hiroyuki
目的：EML 4-ALK是一种肺癌癌基因，ALK抑制剂对携带该融合基因的肿瘤显示出显著的治疗效果。然而，如何在福尔马林固定、石蜡包埋的组织之外的标本中检测融合基因仍然没有解决。我们在此测试基于逆转录PCR（RT-PCR）的EML4-ALK检测是否是一种灵敏可靠的方法。实验设计：我们开发了一种多重RT-PCR系统来捕获ALK融合转录本，并将该技术应用于我们在日本的前瞻性全国性非小细胞肺癌（NSCLC）队列。在2009年2月至12月期间，我们从853例患者中收集了916份标本，对其中754例患者的808份原发性NSCLC标本进行了质量过滤。用我们的RT-PCR系统筛选EML4-ALK和KIF5B-ALK，在来自32个个体（4.24%）的36个样本（4.46%）中鉴定了EML4-ALK转录本。RT-PCR产物在支气管冲洗液（n = 11）、肿瘤活检组织（n = 8）、肿瘤切除组织（n = 7）、胸腔积液（n = 5）、痰液（n = 4）和转移淋巴结（n = 1）中均有表达。结论：多重RT-PCR是检测ALK融合基因转录本的可靠方法。我们建议EML4-ALK的诊断工具应根据可用的标本类型进行选择。FISH和敏感的免疫组化应适用于福尔马林固定，石蜡包埋的组织，但多重RT-PCR是适合于其他标本类型。临床癌症研究; 18（20）; 5682 - 9。(C)2012年AACR。
Purpose: EML4-ALK is a lung cancer oncogene, and ALK inhibitors show marked therapeutic efficacy for tumors harboring this fusion gene. It remains unsettled, however, how the fusion gene should be detected in specimens other than formalin-fixed, paraffin-embedded tissue. We here tested whether reverse transcription PCR (RT-PCR)-based detection of EML4-ALK is a sensitive and reliable approach.Experimental Design: We developed a multiplex RT-PCR system to capture ALK fusion transcripts and applied this technique to our prospective, nationwide cohort of non-small cell lung cancer (NSCLC) in Japan.Results: During February to December 2009, we collected 916 specimens from 853 patients, quality filtering of which yielded 808 specimens of primary NSCLC from 754 individuals. Screening for EML4-ALK and KIF5B-ALK with our RT-PCR system identified EML4-ALK transcripts in 36 samples (4.46%) from 32 individuals (4.24%). The RT-PCR products were detected in specimens including bronchial washing fluid (n = 11), tumor biopsy (n = 8), resected tumor (n = 7), pleural effusion (n = 5), sputum (n = 4), and metastatic lymph node (n = 1). The results of RT-PCR were concordant with those of sensitive immunohistochemistry with ALK antibodies.Conclusions: Multiplex RT-PCR was confirmed to be a reliable technique for detection of ALK fusion transcripts. We propose that diagnostic tools for EML4-ALK should be selected in a manner dependent on the available specimen types. FISH and sensitive immunohistochemistry should be applied to formalin-fixed, paraffin-embedded tissue, but multiplex RT-PCR is appropriate for other specimen types. Clin Cancer Res; 18(20); 5682-9. (C) 2012 AACR.