PMID- 24495999
OWN - NLM
STAT- MEDLINE
DCOM- 20141021
LR  - 20181202
IS  - 1556-1380 (Electronic)
IS  - 1556-0864 (Linking)
VI  - 9
IP  - 3
DP  - 2014 Mar
TI  - Detection of rearrangements and transcriptional up-regulation of ALK in FFPE lung 
      cancer specimens using a novel, sensitive, quantitative reverse transcription 
      polymerase chain reaction assay.
PG  - 307-15
LID - 10.1097/JTO.0000000000000068 [doi]
AB  - INTRODUCTION: The approved dual-color fluorescence in situ hybridization (FISH) 
      test for the detection of anaplastic lymphoma receptor tyrosine kinase (ALK) gene 
      rearrangements in non-small-cell lung cancer (NSCLC) is complex and represents a 
      low-throughput assay difficult to use in daily diagnostic practice. We devised a 
      sensitive and robust routine diagnostic test for the detection of rearrangements 
      and transcriptional up-regulation of ALK. METHODS: We developed a quantitative 
      reverse transcription polymerase chain reaction (qRT-PCR) assay adapted to RNA 
      isolated from routine formalin-fixed, paraffin-embedded material and applied it 
      to 652 NSCLC specimens. The reliability of this technique to detect ALK 
      dysregulation was shown by comparison with FISH and immunohistochemistry. 
      RESULTS: qRT-PCR analysis detected unbalanced ALK expression indicative of a gene 
      rearrangement in 24 (4.6%) and full-length ALK transcript expression in six 
      (1.1%) of 523 interpretable tumors. Among 182 tumors simultaneously analyzed by 
      FISH and qRT-PCR, the latter accurately typed 97% of 19 rearranged and 158 
      nonrearranged tumors and identified ALK deregulation in two cases with 
      insufficient FISH. Six tumors expressing full-length ALK transcripts did not show 
      rearrangements of the gene. Immunohistochemistry detected ALK protein 
      overexpression in tumors with gene fusions and transcriptional up-regulation, but 
      did not distinguish between the two. One case with full-length ALK expression 
      carried a heterozygous point mutation (S1220Y) within the kinase domain 
      potentially interfering with kinase activity and/or inhibitor binding. 
      CONCLUSIONS: Our qRT-PCR assay reliably identifies and distinguishes ALK 
      rearrangements and full-length transcript expression in formalin-fixed, 
      paraffin-embedded material. It is an easy-to-perform, cost-effective, and 
      high-throughput tool for the diagnosis of ALK activation. The expression of 
      full-length ALK transcripts may be relevant for ALK inhibitor therapy in NSCLC.
FAU - Gruber, Kim
AU  - Gruber K
AD  - *Department of Clinical Pathology, Robert-Bosch-Krankenhaus, Dr. Margarete 
      Fischer-Bosch Institute of Clinical Pharmacology, Stuttgart, and University of 
      Tubingen, Germany; daggerInstitute of Pathology, University of Wurzburg, Wurzburg, 
      Germany; double daggerCenter for Pulmonology and Thoracic Surgery, Klinik Schillerhohe, 
      Stuttgart-Gerlingen, Germany; and  section sign Dr. Margarete Fischer-Bosch Institute of 
      Clinical Pharmacology, Stuttgart, and Department of Clinical Pharmacology, 
      University Hospital, Tubingen, Germany.
FAU - Horn, Heike
AU  - Horn H
FAU - Kalla, Jorg
AU  - Kalla J
FAU - Fritz, Peter
AU  - Fritz P
FAU - Rosenwald, Andreas
AU  - Rosenwald A
FAU - Kohlhaufl, Martin
AU  - Kohlhaufl M
FAU - Friedel, Godehard
AU  - Friedel G
FAU - Schwab, Matthias
AU  - Schwab M
FAU - Ott, German
AU  - Ott G
FAU - Kalla, Claudia
AU  - Kalla C
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Thorac Oncol
JT  - Journal of thoracic oncology : official publication of the International 
      Association for the Study of Lung Cancer
JID - 101274235
RN  - 0 (Biomarkers, Tumor)
RN  - 0 (RNA, Messenger)
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  - Biomarkers, Tumor/*genetics/metabolism
MH  - Carcinoma, Non-Small-Cell Lung/*genetics/metabolism/pathology
MH  - Cohort Studies
MH  - Female
MH  - Follow-Up Studies
MH  - *Gene Expression Regulation, Neoplastic
MH  - *Gene Rearrangement
MH  - Humans
MH  - Immunoenzyme Techniques
MH  - In Situ Hybridization, Fluorescence
MH  - Lung Neoplasms/*genetics/metabolism/pathology
MH  - Male
MH  - Middle Aged
MH  - Neoplasm Staging
MH  - Paraffin Embedding
MH  - Prognosis
MH  - RNA, Messenger/genetics
MH  - Real-Time Polymerase Chain Reaction
MH  - Receptor Protein-Tyrosine Kinases/*genetics/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
EDAT- 2014/02/06 06:00
MHDA- 2014/10/22 06:00
CRDT- 2014/02/06 06:00
PHST- 2014/02/06 06:00 [entrez]
PHST- 2014/02/06 06:00 [pubmed]
PHST- 2014/10/22 06:00 [medline]
AID - S1556-0864(15)30212-4 [pii]
AID - 10.1097/JTO.0000000000000068 [doi]
PST - ppublish
SO  - J Thorac Oncol. 2014 Mar;9(3):307-15. doi: 10.1097/JTO.0000000000000068.