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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Greaves, Ronda / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter


IMPACT FACTOR 2018: 3.638

CiteScore 2018: 2.44

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1437-4331
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Volume 51, Issue 9

Issues

Sensitive detection of EML4-ALK fusion oncoprotein of lung cancer by in situ proximity ligation assay

Jin Kyung Rho
  • Asan Institute for Life Sciences, Department of Pulmonary and Critical Care Medicine, Seoul, Korea
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Hyangsin Lee / Chan-Sik Park / Chang-Min Choi
  • Asan Institute for Life Sciences, Department of Pulmonary and Critical Care Medicine, Seoul, Korea
  • Department of Oncology, Asan Medical Center, University of Ulsan, Seoul, Korea
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Jae Cheol Lee
Published Online: 2013-04-09 | DOI: https://doi.org/10.1515/cclm-2013-0044

Abstract

Background: EML4-ALK fusion oncogene has emerged as a novel molecular target in non-small cell lung cancer (NSCLC). Although break-apart fluorescent in situ hybridization (FISH) is the standard method for diagnosis, it is expensive, not readily available and sometimes difficult to interpret. In addition, ALK immunohistochemistry (IHC) may miss the diagnosis because of relatively low level of ALK transcription.

Methods: In situ proximity ligation assay (PLA) originally developed for precise detection and quantification of proteins by dual recognition and amplification process was used for sensitive detection of EML4-ALK fusion oncoprotein in NSCLC cell lines (ALK negative cell: PC-9 and H460, ALK positive cell: H3122 and H2228). EML4-ALK oncogene and protein in lung cancer cells were confirmed by multiplex RT-PCR and Western blots.

Results: We detected 117 kDa variant 1 of EML4-ALK in H3122 and 90 kDa variant 3 of EML4-ALK in H2228. These cells were more sensitive to crizotinib, an ALK inhibitor compared with PC-9 and H460 cells without EML4-ALK rearrangement. After fixing on glass slides by cytospin centrifuge, in situ PLA test was performed. Among four cell lines, distinct, tiny spots were visible only in H3122 and H2228 cell lines with ALK rearrangement. The same results were also obtained when paraffin-embedded cell blocks were used.

Conclusions: Highly specific and sensitive detection of EML4-ALK fusion oncoprotein is possible by in situ PLA method suggesting its clinical application.

Keywords: diagnosis; EML4-ALK; lung cancer

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About the article

Corresponding author: Jae Cheol Lee, MD, PhD, Department of Oncology, Asan Medical Center, College of Medicine, University of Ulsan, 388–1 Pungnap-2 Dong, Songpa-Gu, Seoul 138–736, Korea, Phone: +82 2 30103208, Fax: +82 2 30106961


Received: 2013-01-17

Accepted: 2013-03-18

Published Online: 2013-04-09

Published in Print: 2013-09-01


Citation Information: Clinical Chemistry and Laboratory Medicine, Volume 51, Issue 9, Pages 1843–1848, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/cclm-2013-0044.

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