The Use of Quantitative Real-Time Reverse Transcriptase PCR for 5 ' and 3 ' Portions of ALK Transcripts to Detect ALK Rearrangements in Lung Cancers

	The Use of Quantitative Real-Time Reverse Transcriptase PCR for 5 ' and 3 ' Portions of ALK Transcripts to Detect ALK Rearrangements in Lung Cancers
	Wang, R; Pan, YJ; Li, CG; Hu, HC; Zhang, Y; Li, H; Luo, XY; Zhang, J; Fang, ZY; Li, Y
刊名	CLINICAL CANCER RESEARCH
	2012
卷号	18 期号:17 页码:4725-4732
通讯作者	Chen, HQ (reprint author), Fudan Univ, Shanghai Canc Ctr, Dept Thorac Surg, 270 Dong An Rd, Shanghai 200032, Peoples R China.,sun_yihua76@hotmail.com ; hqchen1@yahoo.com
英文摘要	Purpose: Approximately 3% to 7% of non-small cell lung cancers (NSCLC) harbor an ALK fusion gene, thus defining a tumor group that may be responsive to targeted therapy. The breakpoint in ALK consistently occurs at exon 20 and EML4 or other fusion partners, thus driving a strong expression of ALK kinase domain and resulting in an unbalanced expression in 5' and 3' portions of ALK transcripts. We have developed a rapid and accurate method by simultaneously detecting the expression in 5' and 3' portions of ALK mRNA. Experimental Design: Quantitative real-time reverse transcriptase PCR (qRT-PCR) was used to examine expression levels of the 5' and 3' portions of ALK transcripts in177 NSCLCs, in which EGFR, KRAS, HER2, and BRAF mutations were absent. If unbalanced ALK mRNA expression was seen, ALK rearrangement was assumed to exist. ALK FISH was used to confirm the accuracy of qRT-PCR. RT-PCR and 5' RACE coupling sequencing identified the fusion variants. Results: Real-time RT-PCR showed excellent sensitivity and specificity (100% and 100%, respectively) for detection of ALK rearrangements in resected specimens. In addition, six novel ALK fusion variants were identified, including one KIF5B-ALK (E17;A20) and five EML4-ALK variants (E6a;A19, E6a/b ins 18;A20, E17b ins 39;A20, E10a/b, E13;A20, and E17 ins 65;A20). Conclusions: Real-time RT-PCR is a rapid and accurate method for diagnosing ALK-rearranged lung cancers. Coupling of 5' RACE to this method should further facilitate rapid identification of novel ALK fusion genes. Clin Cancer Res; 18(17); 4725-32. (C) 2012 AACR.
学科主题	Oncology
类目[WOS]	Oncology
关键词[WOS]	ADENOCARCINOMAS ; GENE ; IMMUNOHISTOCHEMISTRY ; MUTATIONS ; KINASES
收录类别	SCI
语种	英语
WOS记录号	WOS:000309964500027
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/677]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Wang, R,Pan, YJ,Li, CG,et al. The Use of Quantitative Real-Time Reverse Transcriptase PCR for 5 ' and 3 ' Portions of ALK Transcripts to Detect ALK Rearrangements in Lung Cancers[J]. CLINICAL CANCER RESEARCH,2012,18(17):4725-4732.
APA	Wang, R.,Pan, YJ.,Li, CG.,Hu, HC.,Zhang, Y.,...&Chen, HQ.(2012).The Use of Quantitative Real-Time Reverse Transcriptase PCR for 5 ' and 3 ' Portions of ALK Transcripts to Detect ALK Rearrangements in Lung Cancers.CLINICAL CANCER RESEARCH,18(17),4725-4732.
MLA	Wang, R,et al."The Use of Quantitative Real-Time Reverse Transcriptase PCR for 5 ' and 3 ' Portions of ALK Transcripts to Detect ALK Rearrangements in Lung Cancers".CLINICAL CANCER RESEARCH 18.17(2012):4725-4732.