PMID- 26973216
OWN - NLM
STAT- MEDLINE
DCOM- 20161213
LR  - 20181202
IS  - 1872-8332 (Electronic)
IS  - 0169-5002 (Linking)
VI  - 94
DP  - 2016 Apr
TI  - A sensitive and high throughput TaqMan-based reverse transcription quantitative 
      polymerase chain reaction assay efficiently discriminates ALK rearrangement from 
      overexpression for lung cancer FFPE specimens.
PG  - 114-20
LID - S0169-5002(16)30224-0 [pii]
LID - 10.1016/j.lungcan.2016.02.004 [doi]
AB  - OBJECTIVES: ALK fusion gene is an oncogenic driver in lung cancer with low 
      prevalence, which can be ameliorated by crizotinib. Currently, ALK fusion gene 
      can be diagnosed by fluorescence in situ hybridization (FISH) and 
      immunohistochemistry (IHC), but inconstistnt results between the two methods are 
      encountered regularly. To make the ALK fusion gene screening more efficient and 
      to provide a simple solution to clarify the discrepancy between FISH and IHC 
      results, a sensitive TaqMan-based reverse transcription quantitative polymerase 
      chain reaction (RT-qPCR) assay was established. MATERIALS AND METHODS: The 3-plex 
      TaqMan-based RT-qPCR assay was established and performed on 102 archived 
      formalin-fixed, paraffin-embedded (FFPE) NSCLC samples to detect ALK 
      rearrangement and overexpression. Break-apart FISH and automatic 
      immunohistochemistry based ALK assays were performed side by side using tissue 
      microarray. RESULTS: The RT-qPCR was performed successfully for 80 samples and 10 
      of them showed positive signals. Three out of the 10 qPCR positive cases were 
      further confirmed by FISH and IHC test. Two others were IHC positive and FISH 
      negative, and expressed full-length ALK transcript. The rest were neither FISH 
      nor IHC positive and their ALK expression level was significantly lower than 
      those FISH or IHC positive cases. CONCLUSION: Our RT-qPCR assay demonstrates that 
      the capability and reliability of ALK detection is comparable to FISH and IHC, 
      but it is more effective at discriminating ALK rearrangement from overexpression. 
      The RT-qPCR assay easily clarifies the discrepancy between FISH and IHC, and can 
      be incorporated into routine ALK screening for lung cancer.
CI  - Copyright (c) 2016 Elsevier Ireland Ltd. All rights reserved.
FAU - Lung, Jrhau
AU  - Lung J
AD  - Department of Medical Research and Development, Chang Gung Memorial Hospital, 
      Chiayi branch, Taiwan.
FAU - Lin, Yu-Ching
AU  - Lin YC
AD  - Division of Thoracic Oncology, Department of Pulmonary and Critical Care 
      Medicine, Chang Gung Memorial Hospital, Chiayi branch, Taiwan; Department of 
      Medicine, College of Medicine, Chang Gung University, Taoyuan, Taiwan; Department 
      of Respiratory Care, Chang Gung University of Science and Technology, Chiayi 
      Campus, Chiayi, Taiwan.
FAU - Hung, Ming-Szu
AU  - Hung MS
AD  - Division of Thoracic Oncology, Department of Pulmonary and Critical Care 
      Medicine, Chang Gung Memorial Hospital, Chiayi branch, Taiwan; Department of 
      Medicine, College of Medicine, Chang Gung University, Taoyuan, Taiwan; Department 
      of Respiratory Care, Chang Gung University of Science and Technology, Chiayi 
      Campus, Chiayi, Taiwan.
FAU - Jiang, Yuan Yuan
AU  - Jiang YY
AD  - Division of Thoracic Oncology, Department of Pulmonary and Critical Care 
      Medicine, Chang Gung Memorial Hospital, Chiayi branch, Taiwan.
FAU - Lee, Kuan-Der
AU  - Lee KD
AD  - Department of Medicine, College of Medicine, Chang Gung University, Taoyuan, 
      Taiwan; Hematology and Oncology, Chang Gung Memorial Hospital, Chiayi branch, 
      Taiwan.
FAU - Lin, Paul Yann
AU  - Lin PY
AD  - Department of Pathology, Chang Gung Memorial Hospital, Chiayi branch, Taiwan.
FAU - Tsai, Ying Huang
AU  - Tsai YH
AD  - Division of Thoracic Oncology, Department of Pulmonary and Critical Care 
      Medicine, Chang Gung Memorial Hospital, Chiayi branch, Taiwan; Department of 
      Respiratory Care, College of Medicine, Chang Gung University, Taoyuan, Taiwan. 
      Electronic address: chestmed@cgmh.org.tw.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20160210
PL  - Ireland
TA  - Lung Cancer
JT  - Lung cancer (Amsterdam, Netherlands)
JID - 8800805
RN  - 0 (Oncogene Proteins, Fusion)
RN  - EC 2.7.10.1 (ALK protein, human)
RN  - EC 2.7.10.1 (Anaplastic Lymphoma Kinase)
RN  - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases)
SB  - IM
MH  - Adult
MH  - Aged
MH  - Aged, 80 and over
MH  - Anaplastic Lymphoma Kinase
MH  - Female
MH  - *Gene Expression
MH  - Gene Expression Profiling
MH  - *Gene Rearrangement
MH  - Humans
MH  - Immunohistochemistry
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Lung Neoplasms/*genetics/pathology
MH  - Male
MH  - Middle Aged
MH  - Neoplasm Staging
MH  - Oncogene Proteins, Fusion/*genetics/metabolism
MH  - Real-Time Polymerase Chain Reaction
MH  - Receptor Protein-Tyrosine Kinases/*genetics/metabolism
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - *Transcriptome
OTO - NOTNLM
OT  - Anaplastic lymphoma kinase
OT  - FISH
OT  - Immunohistochemisty
OT  - Lung cancer
OT  - Real time PCR
EDAT- 2016/03/15 06:00
MHDA- 2016/12/15 06:00
CRDT- 2016/03/15 06:00
PHST- 2016/01/04 00:00 [received]
PHST- 2016/02/03 00:00 [revised]
PHST- 2016/02/06 00:00 [accepted]
PHST- 2016/03/15 06:00 [entrez]
PHST- 2016/03/15 06:00 [pubmed]
PHST- 2016/12/15 06:00 [medline]
AID - S0169-5002(16)30224-0 [pii]
AID - 10.1016/j.lungcan.2016.02.004 [doi]
PST - ppublish
SO  - Lung Cancer. 2016 Apr;94:114-20. doi: 10.1016/j.lungcan.2016.02.004. Epub 2016 
      Feb 10.