ç·šéè Ming SOUND Tsao, MD, FRCPC Fred R. Hirsch, MD, PhD Yasushi Yatabe, MD, PhD
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international Association for the Study of Lung Cancer
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International Association for the Study of Lung Cancer, Aurora, Colorado, USA ç·šéè ïŒ Ming Sound Tsao, MD, FRCPC Fred R. Hirsch, MD, PhD Yasushi Yatabe, MD, PhD IASLC Press ã«ãã£ãŠåºçããã IASLC åºçç© éŠèš³ã«ããã£ãŠæ¬¡ã®æ¹ã ã®ååãããã ããŸãããããŒã¯ãã³ãã³ (Intermed English Services)ãæšæè±ä»£åãè¿è€åæ¶ãæŽç°å žåãå°Ÿé¢é åã倪ç°è£åã è¡šçŽããã³æ¬ã®ã¬ã€ã¢ãŠã㯠Biographics ã«ãããªãªãžãã«ãã¶ã€ã³ IASLC Press ãªãã£ã¹: IASLC, 13100 East Colfax Ave., Unit 10, Aurora, Colorado 80011, USA www.iaslc.org 2013幎10æåç 10 9 8 7 6 5 4 3 2 1 ISBN: 978-1-940488-05-9 èäœæš© c2013 International Association for the Study of Lung Cancer çŠç¡æè»¢èŒ èäœæš©æ³äžã®äŸå€ãé€ããæžé¢ã«ããäºåã®æ¿è«Ÿç¡ãã§åºçç©ã®éšåçå©çšãæ€çŽ¢ã· ã¹ãã ãžã®ä¿åãããã¯å°å ¥ããããã¯å šãŠã®è»¢èŒãçŠæ¢ããŸãã åºçæç¹ã«ãããæ¬æã®æ å ±ã¯çå®ãã€æ£ç¢ºã§ãããšç¢ºä¿¡ããŠããŸããããããªãé éããäžäœçºã«ã€ããŠIASLCãç·šéè ãããã³åºçè ã¯æ³ç責任ãè² ããŸãããåºç è ã¯å 容ã«é¢ããŠæ瀺çã«ãé»ç€ºçã«ãä¿èšŒãè² ããŸããã
èºçã«ãããALKãã¹ã IASLC ã¢ãã©ã¹ ç·šéè
Ming Sound Tsao, MD, FRCPC Fred R. Hirsch, MD, PhD Yasushi Yatabe, MD, PhD
international Association for the Study of Lung Cancer
è¬èŸ IASLC ã¯ãã®ALKã¢ãã©ã¹ã®ãããžã§ã¯ãã«å¯Ÿãã Pfizer Oncology ãæäŸããå¯å€§ãªè³éãšãµããŒãã« æ·±ãæè¬ããŸãããŸã å ±ç·šè ããã³ãã¹ãŠã®æ åœè 㯠Lori Alexander, MTPW, ELS, ã®ç·šéæŽå©ãDeborah A. Whippen, Editorial Rx, Inc., ããã³ Rania Gaspo, PhD, Pfizer Oncology ã®ãµããŒãã«å¯Ÿãæè¬èŽããŸãã
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åæžã . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
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第3ç«
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第4ç«
å ç«çµç¹ååŠæ³ïŒIHCïŒ. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
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é転åé µçŽ ããªã¡ã©ãŒãŒé£éåå¿ïŒRT-PCR)ããã³ãã«ãã ã¬ãã¯ã¹éºäŒå解ææ³ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
第6ç«
ALKãã¹ãã®ããã®è§£æãã©ããããŒã ã®æ¯èŒ . . . . . . . . . . . . . . . . . 44
第7ç«
现è蚺ã«ãããALKãã¹ã . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
第8ç«
ALKãã¹ãã®å ±åæž . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
第9ç«
ã¬ã€ãã©ã€ã³ããã³ãã¹ãæšæºåã®ç 究 . . . . . . . . . . . . . . . . . . . . . . . 61
第10ç«
æŠèŠããã³å±æ. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
åèæç® . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
ä»èš1
èºçã«ãããALKéºäŒååæ§æã«ã€ããŠã®å ±åãããç 究㮠æŠèŠåæ§æ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
ä»èš2
EGFR ããã³ALKããã·ã³ãããŒãŒé»å®³å€ã§æ²»çãåãã èºããæ£è ãéžæããããã®CAP/IASLC/AMP ã®éºäŒå æ€æ»ã®ã¬ã€ãã©ã€ã³ . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
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. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
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List of Contributors ç·šéè Ming Sound Tsao, MD, FRCPC Pathologist and Senior Scientist, Princess Margaret Cancer Centre, University Health Network Professor, Department of Laboratory Medicine and Pathobiology, University of Toronto Toronto, Canada
Fred R. Hirsch, MD, PhD Professor, Department of Medicine Department of Pathology University of Colorado at Denver Denver, Colorado, USA
Yasushi Yatabe, MD, PhD Chief, Department of Pathology and Molecular Diagnostics, Aichi Cancer Center Nagoya, Japan æ åœè ãªã¹ã
åæ å·çè Elisabeth Brambilla, MD, PhD Professor, Département dâAnatomie et Cytologie Pathologiques INSERM U823 Institut Albert Bonniot Centre Hospitalier Universitaire de Grenoble, Université Joseph Fourier Grenoble, France Lukas Bubendorf, MD Professor and Head, Division of Cytopathology Institute for Pathology, University Hospital Basel Basel, Switzerland Jin-Haeng Chung, MD, PhD Professor, Department of Pathology Seoul National University Bundang Hospital Seoul, South Korea
Keith M. Kerr, FRCPath Professor, Department of Pathology Aberdeen University Medical School Aberdeen Royal Infirmary Aberdeen, Scotland, United Kingdom Sylvie Lantuéjoul, MD, PhD Professor and Chair, Département dâAnatomie et Cytologie Pathologiques INSERM U 823 Institut Albert Bonniot Centre Hospitalier Universitair A Michallon, Université Joseph Fourier Grenoble, France Kengo Takeuchi, MD, PhD Pathology Project for Molecular Targets of the Cancer Institute Division of Pathology of the Cancer Institute Hospital Japanese Foundation for Cancer Research Tokyo, Japan
Erik Thunnissen, MD, PhD Consultant Pathologist, VU University Medical Center Amsterdam, The Netherlands Marileila Varella-Garcia, PhD Professor, Department of Medicine/ Medical Oncology Department of Pathology University of Colorado at Denver Denver, Colorado, USA Ignacio Wistuba, MD Professor and Chair, Jay and Lori Eisenberg Endowed Professor Department of Translational Molecular Pathology The University of Texas MD Anderson Cancer Center Houston, Texas, USA Akihiko Yoshida, MD, PhD Attending Pathologist, Department of Pathology National Cancer Center Hospital Tokyo, Japan
2013幎ã€ã¿ãªã¢ã®ãœã¬ã³ãã§éå¬ããã IASLCALKãã¹ãã¯ãŒã¯ã·ã§ããåå è ã åäŸå·ŠããY. Yatabe, M. Varella-Garcia, E. Brambilla, S. Lanteujoul, R. Gaspo ïŒPfizer OncologyïŒ, J-H Chung; åŸåå·Š ããA. Yoshida, I. Wistuba, F. R. Hirsch, M. S. Tsao, E. Thunnissen, L. Bubendorf, K. Kerrã
7
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æ¬æã§äœ¿çšãããŠããç¥èªã AEC: 3-ã¢ãã-9-ãšãã«ã«ã«ããŸãŒã« ALK: æªååãªã³ãè «ãããŒãŒ AMP: Association for Molecular Pathology ATS: American Thoracic Society CAP: College of American Pathologists CISH:ã¯ãã¢ã²ã³ã€ã³ã·ãã¥ãŒãã€ããªãã€ãŒãŒã·ã§ã³ DAB: 3, 3â ãžã¢ãããã³ãžãžã³ EBUS: è¶ é³æ³¢æ°ç®¡æ¯é¡ EDTA: ãšãã¬ã³ãžã¢ãã³åé ¢é ž EGFR: äžç®æé·å åå容äœïŒEpidermal growth factor receptorïŒ EML4: echinoderm microtubule-associated protein-like 4 ERS: European Respiratory Society ETOP: European Thoracic Oncology Platform EUS: çµé£éè¶ é³æ³¢æ³ FDA: ç±³åœé£åå»è¬åå± FFPE: ãã«ããªã³åºå®ãã©ãã£ã³å å FISH: èå ã€ã³ã·ãã¥ãŒãã€ããªãã€ãŒãŒã·ã§ã³ FNA: 现éåžåŒçŽ°è蚺 H&E: ããããã·ãªã³-ãšãªãžã³ HER2: ããäžç®æé·å åå容äœ-2 iAEP: intercalated antibody-enhanced polymer IASLC: International Association for the Study of Lung Cancer IHC: å ç«çµç¹ååŠçæ€æ» ISH: ã€ã³ã·ãã¥ãŒãã€ããªãã€ãŒãŒã·ã§ã³ KIF5B: kinesin family member 5B NOS: Not otherwise specified NSCLC: éå°çŽ°èèºç NGS: 次äžä»£ã·ãŒã¯ãšã³ã·ã³ã° RET: retããéºäŒå ROS1: c-ros ããéºäŒå 1 RT-PCR: é転åé µçŽ ããªã¡ã©ãŒãŒé£éåå¿ SCLC: å°çŽ°èèºç TKI: ããã·ã³ãããŒãŒé»å®³å€ v: å€åœ¢äœïŒvariant)
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以äžã®è£œé æ¥è ããã³è£œåãæ¬ã¢ãã©ã¹ã«èšèŒãããŠããŸããå補é æ¥è ã«ã€ããŠèšèŒãããå Žæ㯠å¯äžã®ãã®ã§ã¯æããŸããã å€ãã®è£œé æ¥è ãäžçåå°ã«ãªãã£ã¹ãæã£ãŠããŸãã
Abbott Molecular Abbott Park, Illinois, USA Vysis LSI ALK Break Apart FISH Probe Kit, Spectrum Orange Probe, and Spectrum Green Probe
Leica Biosystems Buffalo Grove, Illinois, USA Wetzlar, Germany Bond-Max, Novolink Polymer Detection System
Abcam Cambridge, UK Anti-ALK antibodyïŒ5A4ïŒ
Nichirei Biosciences, Inc. Tokyo, Japan 5A4 antibodyïŒHistofine ALK Detection KitïŒ
BDïŒBecton, Dickinson and CompanyïŒ Diagnostics Franklin Lakes, New Jersey, USA SurePath Cell Signaling Technology Danvers, Massachusetts, USA D5F3 antibodyïŒALK [D5F3] XP Rabbit mAbïŒ Dako Glostrup, Denmark Carpinteria, California, USA ADVANCE, ALK1 antibody, EnVision, EnVision+, EnVision FLEX, and EnVision FLEX+, PT Link, and Target Retrieval Solution Hologic, Inc. Bedford, Massachusetts, USA ThinPrep Invitrogen, Life Technologies Corporation Carlsbad, California, USA Anti-ALK antibody
Novocastra Newcastle, UK 5A4 antibody NanoString Technologies Seattle, Washington, USA NanoString assay Pfizer Oncology New York, New York, USA XalkoriTM Ventana Medical Systems, Inc. ïŒmember of the Roche groupïŒ Tucson, Arizona, USA BenchMark XT, iVIEW DAB Detection Kit, OptiView DAB IHC Detection Kit, OptiView Amplification Kit, and ultraView Universal DAB Kit, and Rabbit Monoclonal Primary Antibody assay ZytoVision GmbH Bremerhaven, Germany ZytoDot 2C SPEC ALK break-apart probe
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SIGNAL CLASSIFICATION Patterns observed in native ALK Patterns observed in split 3'-5' ALK
Red and green separated by <2 signal diameters Classified as negative
Red and green separated by â¥2 signal diameters Classified as positive
Red and green separated by â¥2 signal diameters Classified as positive
Red and green separated by <2 signal diameters Classified as negative
ã¹ããªããïŒéœæ§ïŒãã¿ãŒ ã³ïŒå³ 7BïŒãïŒã€ã®çŽ°è ã«ãããŠã5'ãš 3'ã®ã·ã°ã ã«ãåé¢ããŠååšããŠãã SPECIMEN CLASSIFICATION Nonrearranged tumors: Rearranged tumors: ãšããéé¢ã·ã°ãã«ã®æ°ã« Rearrangement-positive cell rate Rearrangement-positive cell rate ãããããã ã¹ããªãã <15% of cells â¥15% of cells ãã¿ãŒã³ïŒALKéœæ§ïŒãš å³ 6ããã¬ãŒã¯ã¢ããŒã FISH ã«ããALKèåèºè «ç现èæ žã®ã·ã°ãã«ãã¿ãŒã³ã 解éãããïŒå³3ïŒã5'ãš 3'ã®ã·ã°ãã«ã®åé¢ã®è·é¢ã¯ãæã倧ããã°ãã«ã®çŽåŸã® 2 å以äžã§ãªããã°ãªããªã ïŒCamidge 2010ïŒã5'ãš 3'ã®åé¢ã·ã°ãã«ã®æ°ãçããå¿ èŠã¯ãªããäŸãã°ã5'åŽã®ã· ã°ãã«ã®ã³ããŒæ°ã 2ã€ã§ããã㊠3'åŽã®ã·ã°ãã«ã®ã³ããŒæ°ã 3ã€ã®çŽ°èã¯ã¹ããªãã ãšããŠåé¡ããããïŒçŽ°èåœããã® 5'-3' Patterns Examples èåã·ã°ãã«æ°ã¯ãã¿ãŒã³åé¡ã«é¢äžã ãªãã åäž 3âïŒéœæ§ïŒãã¿ãŒã³ïŒå³ 7CïŒãå äž3'ã·ã°ãã«ããåäž5'ã·ã°ãã«ç¡ã㧠æ€åºãããå Žåã现èã¯åäž3'ãã¿ãŒã³ ãæãããšè§£éããããïŒã€ã®çŽ°èã§å äž3'ã·ã°ãã«ãš åäž5'ã·ã°ãã«ã®äž¡æ¹ã æãã3'ã·ã°ãã«ã 5âã·ã°ãã«ããå€ã å Žåãæ£ããåé¡ã¯ã¹ããªãããã¿ãŒã³ ã§ãåäž3'ã§ã¯ãªããã·ã°ãã«èåæ°ã¯ ãã¿ãŒã³åé¡ã«é¢äžããªãã
éé¢ 5âïŒé°æ§ïŒãã¿ãŒã³ïŒå³7DïŒãå äž5'ã·ã°ãã«ããåäž3'ã·ã°ãã«ç¡ã㧠æ€åºãããå Žåã现èã¯åäž5'ãã¿ãŒã³ ãæãããšè§£éãããïŒALKé°æ§ïŒã现 èãåäž3'ãš åäž5âã·ã°ãã«ã®äž¡æ¹ãæ ãã5'ã·ã°ãã«ã 3'ã·ã°ãã«ããå€ã å Žåãæ£ããåé¡ã¯ã¹ããªãããã¿ãŒã³ ã§ãåäž 5âã§ã¯ãªããã·ã°ãã«èåæ°ã¯ ãã¿ãŒã³åé¡ã«é¢ä¿ãªãã
A. Fused
B. Split
C. Isolated 3' ALK
D. Isolated 5' ALK å³ 7ãFISH ã«ãããALKã·ã°ãã«ãã¿ãŒã³ã«åºã¥ãè «ç 现èã®åé¡ã
22
èºçã«ãããALKãã¹ãIASLC ã¢ãã©ã¹
泚: ã¹ããªãããã¿ãŒã³ã®å€å®èŠæºã¯ Vysis LSIALKBreak Apart FISH Probe Kit ãçšã ã FFPE è «çåçã®ãã¹ãã«åºã¥ãããã®ã§ãç°ãªã解æè©Šè¬ãããã¯çäœæšæ¬ã䜿çšãã ãå Žåã«ã¯ãã®è©äŸ¡èŠæºã®åŠ¥åœæ§ã確èªããå¿ èŠãããããããŒãã®ãµã€ãºã¯ãããŒãã®ã ã¶ã€ã³ã«ãã£ãŠç°ãªãå Žåããããã·ã°ãã«ãµã€ãºã®å€§ãããããŒãã䜿çšããå Žåã ã ãå°ããåé¢è·é¢ãã¹ããªããã®å®çŸ©ã«å¿ Fused 5' 3â èŠãšãªãã Cell Pattern Cell 1
ã¹ã³ã¢ãªã³ã°
ã¹ã³ã¢è ãäžäººã®å Žåã¯æäœ 50è «ç现è ããã¹ã³ã¢è ãäºäººã®å Žåã¯æäœ 100 ã® è «ç现èãå¿ èŠãšãããŠããïŒè©³çŽ°ã«ã€ ããŠã¯ãã«ãããªãå€ãã®é ãåç §ïŒã è©äŸ¡å¯èœãªçŽ°èæ°ãå°ãªãæšæ¬ã¯ FISH 解æã«é©ããªãïŒCamidge 2010ïŒãå è «ç现èã«ã€ããŠã®ã·ã°ãã«ãã¿ãŒã³ã ã¹ã³ã¢ãªã³ã°ã®ã¯ãŒã¯ã·ãŒãã«èšé²ãã¹ ãã§ããïŒå³ 8ïŒãã¹ã³ã¢ãªã³ã°ã¯åäž ïŒèµ€ãšç·ïŒããã³äºéå¹²æžãã£ã«ã¿ãçšã ãŠçµç¹ãè©äŸ¡ãããšãããæ£ç¢ºã§ããã ç»åã«åºã¥ããã¹ã³ã¢ãªã³ã°ã«ã¯é床ãã ããåé¢ã·ã°ãã«ã®èª€ã£ã 解éãé¿ããããã«ã¯ãç» åã¯ãã¹ãŠã®åç深床ã代 è¡šãããã®ã§ãªããã°ãªã ãªãïŒå³9ïŒã æ£åžžALKéºäŒåã®ã³ã㌠æ°å¢å 㯠NSCLCïŒå³ 10ïŒ ã§ã¯ããèªããããèçœè³ª ã®éå°çºçŸãšã®é¢é£æ§ã¯ç€º ãããŠããªããçŸæç¹ã§ ã¯ãALKã®ã³ããŒæ°ãã«ãŒ ãã³ã«ã¹ã³ã¢ãªã³ã°ããã ãšã¯æšå¥šãããŠããªãã
A
signal
signal
signal
2
0
0
Fused
Cell 2
2
1
1
Split
Cell 3
2
0
1
Isolated 3'
Cell 4
1
1
0
Isolated 5'
Cell 5
0
1
2
Split
Cell 6
1
0
0
Fused
Cell 50
2
0
0
Fused
Summary of Scoring: Total # of cells scored: 50 Total # of cells with fused pattern: 19 Total # of cells with split pattern: 22 Total # of cells with isolated 3' pattern: 5 Total # of cells with isolated 5' pattern: 4 Total # of cells with rearrangement-positive patterns: 22+5=27 Rearrangement-positive cell rate: 27÷50 x100=54%
å³ 8ã现èã¹ã³ã¢ãªã³ã°çšã¯ãŒã¯ã·ãŒãã®äŸã
No z-stacking
B
16 planes at 0.4 µm= 6.4 µm z-stacking
å³ 9ãç»åã«åºã¥ãã解æ㯠z 軞ã«æ²¿ãéå±€ïŒäžå¹³é¢äžãžã®è€æ°çŠç¹ã¬ãã« ã®çµ±åïŒãåçã®å šæ·±åºŠã代衚ããªãå Žåã¯å°é£ã§ããã
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36
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å³ 11ã KIF5B-ALK- éœæ§è ºçã® IHC ã«ãããç°åžžãªéœæ§åå¿ããŽã«ãžéã匷調ãããããªåŒ·æè²ã ã¿ãŒã³ã瀺ãïŒA: H&E æè², B:ALKæè²ïŒãããã³æ žåšå²ã®ãããŒãã¿ãŒã³ïŒC: H&E æè², D:ALK æè²ïŒãã㌠= 100 ÎŒmãTakeuchi K, et al. Clin Cancer Res. 2009;15: 3143-3149. ãã転èŒã
37
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39
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del49e20
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EML4-ALK (E15del19;del20A20), v4'
e20
e6
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EML4-ALK (E13;ins69A20), v6
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e20
e13
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del14e20
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å³ 1ãéçåALKããã³èºçã«ãããããŸããŸãªã¿ã€ãã®ALKèå: 5 ã€ã®éåžžèªããããã ãªã¢ã³ã(v1ãv2ã v3ãv5 ããã³ v5')ãš 4 ã€ã®å€åçãªEML4-ALKã®ããªã¢ã³ã(v4ãv6 ãv7ãããã³ v4')ã3 ã€ã®KIF5B-ALKããªã¢ã³ã ãããã³ KLC1-ALKãèåããŒãããŒéº äŒå (EML4, KIF5B, and KLC1) ã®æ¿ãéšåã¯ã³ã€ã«ãã³ã€ã« ãã¡ã€ã³ãè¡šããALKå ã§ã¯ ãããã¯è貫éãã¡ã€ã³ïŒãªã¬ã³ãžè²ïŒãšãããŒãŒãã¡ã€ã³ïŒèµ€ïŒãè¡šãã
5BïŒKIF5B) ããã³ kinesin light chain 1ïŒKLC1ïŒãã¯æ€åºã§ããªãïŒTakeuchi 2009ã Togashi 2012ïŒããã®éçãå æããããã«ãä»ã®èåããŒãããŒã®ããã®ãã©ã€ããŒã
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40
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è¡š 1ã EML4-ALKã¹ã¯ãªãŒãã³ã°ã®ããã® RT-PCR ã®èšå®
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é »ç¹ã«èªããããEML4-ALKããªã¢ã³ããžã®é©çšå¯ èœæ§ äž»èŠãªããªã¢ã³ã*
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E20;A20
E6;A20
E2;A20
E18;A20
E21;A20
Soda et al., 2007
Yes
Yes
No
No
Yes
Yes
åçµ
2
Inamura et al., 2008
Yes
Yes
No
No
Yes
Yes
åçµ
2
Shinmura et al., 2008
Yes
Yes
No
No
Yes
Yes
åçµ
3
Takeuchi et al., 2008
Yes
Yes
Yes
Yes
Yes
Yes
åçµ
3
Martelli et al., 2009
Yes
Yes
Yes
No
Yes
Yes
åçµ
3
Takeuchi et al., 2009
Yes
Yes
Yes
Yes
Yes
Yes
åçµ
(KIF5B-ALKã®ããã® 2ã€ã®ãã©ã€ããŒãå«ã)
Wong et al., 2009
Yes
Yes
Yes
No
Yes
Yes
åçµ
4
Takahashi et al., 2010
Yes
Yes
Yes
Yes
Yes
Yes
åçµ
3
Sun Y et al., 2010
Yes
Yes
Yes
Yes
Yes
Yes
åçµ
4
Sanders et al., 2011
Yes
Yes
Yes
Yes
Yes
Yes
FFPE
23
5
6
Takeuchi et al., 2012
Yes
Yes
Yes
Yes
Yes
Yes
åçµ
Shaozhang et al., 2012
Yes
Yes
Yes
Yes
Yes
Yes
åçµ
4
Soda et al., 2012
Yes
Yes
Yes
Yes
Yes
Yes
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(KIF5B-ALKã®ããã® 4ã€ã®ãã©ã€ããŒãå«ã)
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9
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41
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EML4 72F/ !"#$(2-HN( &#'(+.26AA( ALK 3078RR
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KIF5B867F 5'-ATTAGGTGGCAACTGTAGAACC-3' (exon 10) 'OH3P672H(3@%&BB&DDBDDC&&CBDB&D&&CC%+@(();EF>(*./( KIF5B2533F'OH3P-3++H((3@%DBDC&C&&&C&DBBDDB&CDBD%+@()!EF>(-+%-$(4FI>:MGQ/( 5'-GTGCACAAACAGTTGGTACGTG-3' (Exon 23-24 boundary)
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3 ã€ã®ç 究㧠D5F3 ã¯ããŒã³ãçšãã IHC ãš FISH ãæ¯èŒãããïŒè¡š 2ïŒïŒMinoKenudson 2010, Martinez 2013, Minca 2013ïŒãæåæ§ã¯ 83ïœ100% ã§ãããç¹ç°æ§ 㯠99% ããã³ 100% ã§ãã£ãããã®äžã®ç 究ã®1ã€ã§ã¯ãFISHã§ALKéœæ§ã§ãã£ã 6 æš æ¬ã®ãã¡ã1 æ€äœã¯ IHC ã«ãã£ãŠALKé°æ§ã§ãã£ãããFISH ã«ãã£ãŠALKé°æ§ã§ãã£ãæš æ¬ã§ IHC ã§ALKéœæ§ã®ãã®ã¯ç¡ãã£ãïŒMartinez 2013ïŒããããã®ç 究ã«å ããŠã åœé çãªç çå»ããã«ã¯ãæ¢ç¥ã®ALKéºäŒååã«ããäžé£ã®èºè ºçã D5F3 IHC ãçšããŠè©äŸ¡ ãã ãå šäœçã«ã¿ãŠã FISH ãšæ¯èŒãã IHC ã®æåæ§ã¯ 90% ã§ãç¹ç°æ§ã¯ 95% ã§ã ãã芳å¯è éã®ã¹ã³ã¢ãªã³ã°ã®äžèŽçã¯é«ãã£ãïŒHirsch 2013ïŒã è¡š 2ãD5F3 æäœãçšãã IHC æè²ã®ç¹åŸŽ
ç 究
MinoKenudson et al., 2010
Martinez et al., 2013
Minca et al., 2013a Hirsch et al., 2013b* a
æšæ¬ã® æšæ¬ã® æ° æ°
æå 賊掻åæ³
æ€åºãš å¢å¹ ã·ã¹ãã
IHC éœæ§ éŸå€
IHC æåæ§ (察 FISH)
IHC ç¹ç°æ§ (察 FISH)
0察+
â¥10% éœæ§ è «ç现è
83%
100%
ã¹ã³ã¢ãªã³ã°
153
1:100
EDTA (pH 8.0) å§åé
EnVision+
0ã1+ã2+ã >10% ãŸã㯠éœæ§ 3+ ããã³ è «ç现è è «ç现è %
100%
99%
79
1:50
æšæº BenchMark XT
ultraView
231
1:100
æž©åºŠèª¿æŽ BenchMark XT
OptiView
0察+
éœæ§
94%**
100%
98
ååžé
æšæº BenchMark XT
OptiView ãš OptiView å¢å¹
0察+
匷ã 现è質 æè²
90%
95%
FFPE æšæ¬ãçšãã IHC æ€æ»ã® 2 ã€ã®åœé°æ§çµæ㯠ThinPrep ããã»ããµãŒ(éœæ§ã®çµæ)ãçšããæ€èšã§ä¿®æ£ãããã
b
ç 究ã«éžã°ããçäŸã®ãã¡ã43 æ€äœã¯ FISHãã¹ãã«ãã£ãŠALKéœæ§ã§ã55 æ€äœã¯ FISH ã«ãã£ãŠALKé°æ§ã§ãã£ãã
BenchMark XT, ultraViewïŒUniversal DAB Detection KitïŒ, OptiViewïŒDAB IHC Detection KitïŒ, ããã³ OptiView Amplification Kit 㯠Ventana Medical SystemsãInc.ã®è£œåã EnVision+ 㯠Dako ã®è£œåã
5A4 IHC 察 FISH
å°ãªããšã 6 ã€ã®å€§ããªã·ãªãŒãºã®ç 究㧠5A4 ã¯ããŒã³ãçšãã IHC ãš FISH ãæ¯èŒã ããïŒè¡š 3ïŒïŒJokoji 2010, Kim 2011, Paik 2011, Lopes 2012, McLeer-Florin 2012, Sholl 2013ïŒããã¹ãŠã®ç 究ãéããŠãæåæ§ããã³ç¹ç°æ§ã¯ãããã 93ïœ100% ãŸã§, ããã³ 96ïœ100% ã«åãã ããããããã®äžã®ç 究ã§ãåœéœæ§ãããã¯åœé°æ§ã®çµæãå ± åãããŠããã1 äŸã¯ãæªæ§è «ç现èã FISH ã§æ€åºããããçµæ㯠IHC ALKéœæ§ã§ãã£
47
第 6 ç« : ALKãã¹ãã®ããã®ç°ãªã£ã解æãã©ããããŒã ã®æ¯èŒ
è¡š 3ã5A4 æäœãçšãã IHC æè²ã®ç¹åŸŽ ç 究
5A4 æšæ¬ åžéå æ° ç
æå 賊掻åæ³
æ€åºãš å¢å¹ ã·ã¹ãã
254
1:100
æšç賊掻åæ³ æº¶æ¶²ã é«ãPH
EnVision FLEX+ ã ãã³ IAEP
Paik et al. 2011
640
1:30
CC1 溶液
i-VIEW
Kim et al., 2011
465
1:30
CC1 溶液
i-VIEW
McLeerFlorin et al., 2012
441
1:50
CC1 溶液
ultraView
Lopes et al., 2012
62
1:200
é»åã¬ã³ãž
Novolink
Sholl et al., 2013
186
1:50
ã¯ãšã³é žç·©è¡ 液ã(pH 6) å§åéäž
EnVision FLEX+
Jokoji et al., 2010
IHC éœæ§ éŸå€
ã¹ã³ã¢ãªã³ã° 0察+
ã¹ã³ã¢ç¡ã
IHC æåæ§ (察 FISH) 100%
(FISH 㯠IHC+ æšæ¬ ã§ã®ã¿è¡ ãããïŒ
IHC ç¹ç°æ§ (察 FISH) 100%
100%
96%
100%
98%
>10% è «ç现è éœæ§
95%
100%
ã¹ã³ã¢ç¡ã
>10% è «ç现èéœæ§
100%
100%
0ã1+ããŸã 㯠2+
1+, 2+
93%
100%
0ã1+ã2+ã 0ããŸã㯠1+: ãŸã㯠3+ é°æ§ 2+: é°éœäžæ 3+: éœæ§
0ã1+ã2+ã 0ããŸã㯠1+: ãŸã㯠3+ é°æ§ 2+: é°éœäžæ 3+: éœæ§
0ã1+ã2+ã ãŸã㯠3+ ãã ã³ è «ç现è éœæ§ ïŒ
ã¹ã³ã¢ã 2+ ã®å Žå,éœæ§ãš å€å®
a McLeer-Florin et al., Jokoji et al., ããã³ Lopes et al.ã®ç 究ã«äœ¿çšããã 5A4 æäœã¯ Abcam ã®è£œå; ä»ã®ç 究ã§äœ¿çšãããæäœã¯ Novocastra ã®è£œåã iView (DAB Detection Kit) ããã³ OptiView (DAB IHC Detection Kit) 㯠Ventana Medical Systems ã®è£œåãNovolink (Polymer Detection System) 㯠Leica Biosystems ã®è£œåã
ããå¥ã®äŸã§ã¯ã匱ãç·ã®ãããŒãã·ã°ãã«ããã€é察称çãªã¹ããªããã·ã°ãã«ã FISH ALKéœæ§ãšèª€ã£ãŠè§£éãããŠããïŒSholl 2013ïŒãå šäœçã«ã¿ãŠããã®ç 究ã§ã¯ãFISH ãš æ¯èŒãã IHC ã®ç¹ç°æ§ã¯ 98.5% ã§ãã£ãã ãŠãµã®ããªã¯ããŒãã«æäœ IHC 察 RT-PCR
å°ãªããšã 2 ã€ã®ç 究ã§ã¯ããŠãµã®ã®ããªã¯ããŒãã«æäœãçšãã IHC ãè¡ãããŠãã ïŒè¡š 4ïŒïŒWong 2009, Chen 2012ïŒããã®å ã®ïŒã€ã®ç 究ã§ã¯ãçµæ㯠EML4-ALK RT-PCR ã§åŸãããçµæãšæ¯èŒãããäžèŽçã¯é«ãã£ãïŒChen 2012ïŒãä»ã®ç 究ã§ã¯ãRTPCR ã§ALKéœæ§ã® 11 ã®è ºçãš 1 ã€ã®è ºæå¹³äžç®çã IHC ã§ãéœæ§ãšå€å®ãããïŒWong et al 2009ïŒã è¡š 4ãALKãŠãµã®ããªã¯ããŒãã«æäœãçšãã IHC æè²ã®ç¹åŸŽ
ç 究
Wong et al., 2009 Chen et al., 2012
æšæ¬ã®æ° (ALK+ ALK1 ã® %) åžéåç 266 (4.9%)
64 (4.7%)
æå 賊掻åæ³
1:1000 ã¯ãšã³é žç·©è¡æ¶² (pH 6.0) é»åã¬ã³ãž 1:500
CC1 溶液
æ€åºãš å¢å¹ ã·ã¹ãã
HRP ã³ã³ãã¬ãã¯ã¹ ultraView
ã¹ã³ã¢ãª ã³ã°
IHC éœæ§ éŸå€
ç¡ã
ç¹å®ç¡ã
0ã1+ã 2+ã ãŸã㯠3+
â¥2+
IHC IHC æåæ§ ç¹ç°æ§ (察 RT-PCR) (察 RT-PCR) 100%
ç¹å®ç¡ã
100%
90%
CC1 (TTris/borate/ EDTA) 㯠Roche (ããŒãŒã«ãã¹ã€ã¹é£éŠå ±ååœ)ã®è£œåã§ãultraView (Universal DAB Detection Kit) 㯠Ventana Medical Systems, Inc. , ãã㊠HRP (horseradish peroxidase) complex 㯠Dako ã®è£œåã
48
èºçã«ãããALKãã¹ãIASLC ã¢ãã©ã¹
è¡š 5ãIHC ã§äœ¿çšãããæäœã¯ããŒã³ã®æ¯èŒ
ç 究
Takeuchi et al., 2009
Rodig et al., 2009
æšæ¬ã®æ° (ALK+ æäœ ã® %) ã¯ããŒã³
21 (52%)
239 (4%)
361a (5%)
æšç賊掻å 溶液ïŒpH 9.0) ïŒå šãŠ)
Selinger et al., 2013
377 (3%)
594 (11%)
iAEP
ã¹ã³ã¢ãªã³ã° 0察+ (å šãŠ)
æšæº
RT-PCR
IHC IHC æåæ§ ç¹ç°æ§ (察 æšæº) (察 æšæº) 100%
100%
5A4
EnVision+
27%
100%
iAEP
100%
100%
ALK1
EnVision+
9%
100%
SP8
iAEP
20%
100%
SP8
EnVision+
100%
18%
40%
100%
80%
100%
67%
97%
100%
99%
81%
100%
EnVision FLEX+
100%
100%
EnVision FLEX+
100%
99.7%
100%
62.5%
ALK1
ALK1
EDTA å§åéäžïŒäž¡ æ¹ïŒ
EnVision+
EDTA å§åéäžïŒäž¡æ¹
EnVision+ (äž¡æ¹)
ãã©ãã å¢å¹
D5F3 ALK1
æšç賊掻åæ³ æº¶æ¶²ïŒpH 9.0) (å šãŠ)
5A4
Conklin et al., 2013
æ€åºãš å¢å¹ ã·ã¹ãã
ALK1
ALK1
37 (59%) MinoKenudson et al., 2010 Murakami et al., 2011
5A4
æå 賊掻åæ³
5A4
補é å 㮠䜿çšèª¬æã« æºæ ïŒå šãŠïŒ
ABC (å¢å¹ ç¡ã)
iAEP
0察 + (äž¡æ¹)
FISH
ã¹ã³ã¢ >2.7 ç»åäž è§£æ㯠éœæ§
FISH
0察+ (å šãŠ)
RT-PCR/ FISH
0ã1+ã2+ã ãŸã㯠3+ (å šãŠ)
FISH
ALK1
EnVision FLEX
66%
100%
ALK1
ADVANCE
66%
87.5%
5A4
ADVANCE
100%
87.5%
D5F3
ADVANCE
100%
75%
100%
99%
0ã1+ã2+ã ãŸã㯠3+ (å šãŠ)
FISH
ALK1
ç·©è¡æ¶²ïŒpH 9.0) å§åéäž
EnVision FLEX+
5A4
補é å 㮠䜿çšèª¬ææžã« æºæ
ultraView ãš ultraView å¢å¹
100%
98%
D5F3
補é å 㮠䜿çšèª¬ææžã« æºæ
OptiView ãš OptiView å¢å¹
100%
99%
a D5F3 æäœæ€æ»ã®ããã®æšæ¬æ°ã¯ 356 ã§ãã£ãã SP8 㯠Abcam ã®è£œåãTarget Retrieval Solution, buffer, EnVision, EnVision FLEX+, ããã³ ADVANCE 㯠Dako ã®è£œåãã§ã ããultraView (Universal DAB Detection Kit) , OptiViewïŒDAB IHC Detection Kit), ããã³ ultraView and OptiView Amplification
kits 㯠Ventana Medical SystemsãInc.ã®è£œåã§ããã ABC=avidin biotin complex.
49
第 6 ç« : ALKãã¹ãã®ããã®ç°ãªã£ã解æãã©ããããŒã ã®æ¯èŒ
æäœã¯ããŒã³ã®æ¯èŒ
åããµã³ãã«ã»ããã䜿çšããŠæäœã¯ããŒã³éã®äžèŽçãæ€èšããç 究ããããäžèŽçã¯æ äœã«ãã£ãŠç°ãªã£ãŠããïŒè¡š 5ïŒïŒRodig 2009, Takeuchi 2009, Mino-Kenudson 2010, Murakami 2011, Conklin 2013, Selinger 2013ïŒãTakeuchi et al.ã¯ã·ã°ãã«å¢å¹ ã®ãã ã«æäœå¢åŒ·ããªããŒã®æ¿å ¥ïŒiAEPïŒæ¹æ³ãçšããŠALK1 ããã³ 5A4 æäœã®çµæã®åçæ§ãå ± åããïŒTakeuchi 2009ïŒããããã äžäžèŽãèªããããæ€èšããããäŸãã°ãMurakami et al. ã¯ALKåæ§æãã〠12 ã®æšæ¬é㧠1 ã€ã®äžäžèŽäŸãå ±åããããã®çµæ㯠5A4 IHC ã§é°æ§ã§, D5F3 IHC ã§éœæ§ã§ãã£ãïŒMurakami 2012ïŒãConklin et al. 㯠æäœã¯ã㌠ã³ãšæ€åºã·ã¹ãã ã® 5 ã€ã®çµåããæ¯èŒãããADVANCE systemïŒDakoïŒãçšããå Žå 5A4 ãš D5F3 ãæãé«ãäžèŽçã瀺ãããã FISHã§ALKé°æ§ã ã£ãæšæ¬ã§ã¯äžåäžãªéœ æ§åå¿ãæ€åºãããïŒConklin 2013ïŒã
ãã«ããã¬ãã¯ã¹ãããã¯å®éç㪠RT-PCR 察 FISHïŒIHC ã®æç¡ãåããïŒ
ããç 究ã§ã¯ãEML4-ALK転åç©ã 9 人㮠NSCLC æ£è ããæ¡åããè «ç现èã§èªããã ããããæ£åžžãªèºã§ãèªããããïŒMartelli 2008ïŒãæ£åžžãªèºçµç¹ã«ãã®ãããªè»¢åç©ãèª ããããããšã¯ä»ã®ç 究è ã«ãã£ãŠè¿œèªãããŠããªããããããŒã¿ã«çããæãããïŒMano 2010, Sasaki 2010ïŒãRT-PCR ã¯é«ãç¹ç°æ§ãšæåæ§ããã€æ¹æ³ã§ãåœéœæ§ã¯èªããã㪠ããããããFFPE æšæ¬ããè¯è³ªã® RNA ãååŸããããšãé£ãããã,åœé°æ§ãããããå± éºããããRT-PCR ã®åœé°æ§çã«ã€ããŠè©³ããå ±åã¯ç¡ãããååãç 究ã§ã®ALK転åç©ã® æ€åºæåãå ±åãããŠããïŒSoda 2012ïŒããã®ç 究ã§ã¯ã916 ã®æ€äœäž 108ïŒ12%ïŒã¯ RNAãå解ããŠãã解æããé€å€ããããEML4-ALK転åç©ã¯ 36 æ€äœã§æ€åºããããã®ã ã¡ 15 ã« IHC ãæœè¡ã§ãã 15äŸãã¹ãŠãALK IHCéœæ§ã§ãã£ãã äžè¬ã«ãALK転åç©æ€åºã®ããã® RT-PCR ã®æåæ§ãšç¹ç°æ§ã¯IHC ããã³ FISH ãšæ¯èŒã ãŠæºè¶³ã§ãããã®ã§ããã94ïœ100% ã«ãããã§ããïŒè¡š 6ïŒïŒTakeuchi 2008, Inamura 2008, Takeuchi 2009, Soda 2013ïŒã è¡š 6ãä»ã®ALKãã¹ãæ¹æ³ã®çµæãš RT-PCR çµæã®æ¯èŒ
ç 究
ã¿ã€ã RT-PCR
IHC æäœ
FISH
Inamura et al., 2008
ãã«ããã¬ã㯠ã¹RT- PCR
ALK1
Takeuchi et al., 2008
ãã«ããã¬ã㯠㹠RT- PCR
â
Takeuchi et al., 2009
ã€ã³ããŒã¹ãã ã³ ãã«ããã¬ã㯠ã¹RT- PCR
Soda et al., 2012
ãã«ããã¬ã㯠ã¹RT- PCR
ç¡ã
RT-PCR ã®æåæ§ RT-PCR ã®ç¹ç°æ§ (察 FISH ããã³/㟠(察 FISH ããã³/ ã㯠IHC) ãŸã㯠IHC) 100% (察 IHC)
100% (察 IHC)
FISH-ããŒã¹ã®èå解æ
100% (察 FISH)
100% (察 FISH)
5A4 ALK1
EML4 ãã㊠KIF5B
100% (察 IHC)
100% (察 IHC)
5A4
ALKãã¬ãŒã¯ã¢ããŒã ãããŒãããã
100% (察 IHC) 94% (察 FISH)
100% (察 FISH) 100% (察 FISH)
èå解æ
Inamura et al. ããã³ Takeuchi et al.(2009) ã«ããç 究ã«çšããããALK1 æäœã¯ Dako 補åã§ãããTakeuchi et al.(2009) ã®ç 究ã§
䜿çšããã5A4 æäœã¯ Abcam ã®è£œåã§ã Soda et al.ã®ç 究ã§äœ¿çšãããç©ã¯ Nichirei Biosciences, Inc. ã®è£œåã§ãããSoda et al.ã® ç 究ã§äœ¿çšããããã¬ãŒã¯ã¢ããŒããããŒãããã㯠Abbott Molecular ã®è£œåã§ããã
50
èºçã«ãããALKãã¹ãIASLC ã¢ãã©ã¹
CISH 察 ä»ã®æ¹æ³
CISH ã«ã¯ FISH ã«åãå©ç¹ããããALKåæ§ææ€åºã«ããã CISH ã®çµæãä»ã®æ¹æ³ã®ããã ãšåçã§ããããšã瀺ãããŠããïŒè¡š 7ïŒïŒKim 2011, Yoshida 2011a, Schildhaus 2013ïŒã CISH ã®ããã®ALKéœæ§ã®èŠæºã¯ FISH ã®åºæºãšåãå ±åãããã°ã CISH ã®ããã«å¥ã®ã«ã ããªãå€ãçšããäºã«ãããALKéœæ§ãšALKé°æ§ã®è «çãããè¯ãåããããšãã§ãããšãã å ±åãããïŒSchildhaus 2013, Yoshida 2011aïŒãæ°ããæè¡ãéçºãããALKã³ããŒæ°ã ãã³ALKèçœè³ªè¡šçŸãåäžçŽ°èã§è©äŸ¡ãããæè¡ãããïŒVentana Medical SystemsãInc.ïŒ ïŒVentana Medical SystemsãInc.ïŒããã®æ¹æ³ã¯ããæ£ç¢ºãªALKç¶æ ã瀺ããåŸæ¥ã® IHC ãããåªããæåæ§ã蚌æããŠããããããããã®è§£æã¯æè¡çã«é£ããããã®æè¡ã®æ€èšŒ äœæ¥ãé²è¡ããŠããã è¡š 7ãALKãã¹ãã®ä»ã®æ¹æ³ãš CISH ã®æ¯èŒ CISH
ç 究
解æããã
Kim et al., ALKäºè² 2011 ãã¬ãŒã¯ã¢ããŒã CISH Yoshida et al., 2011a
ALKäºè² ãã¬ãŒã¯ã¢ããŒã CISH
Schildhaus ZytoDot 2C SPEC et al., ALKãã¬ãŒã¯ã¢ã 2013 ãŒããããŒã
æšæ¬ã®æ° (ALK+ ã® %)
éœæ§ã® åºæº
431 (4%)
èšè¿°ç¡ã
FISH
IHC
RT-PCR
æåæ§ ç¹ç°æ§ æåæ§ ç¹ç°æ§ æåæ§ ç¹ç°æ§ 94%
100% 66.7% 100%
NA
NA
45 (31%)
â¥20% 100% 100% ã¹ããªãã ã·ã°ãã«äŒŽã
93%
100%
93%
100%
100 (16%)
â¥15% ã®çŽ°è 100% 100% ã¹ããªãã ã·ã°ãã«äŒŽã
NA
NA
NA
NA
Kim et al. ããã³ Yoshida et al.ã®ç 究ã«äœ¿çšãã解æããã㯠Ventana Medical SystemsãInc.ã®è£œåã Schildhaus et al. ãç 究ã§äœ¿çš ããããã㯠Gmbh ZytoVision ã®è£œåã NA =å©çšã§ããªãã
ALKãã¹ãã®ããã®èšºæã¢ã«ãŽãªãºã ã®äŸ
ALKæ€åºã®ããã®åã ã®æ¹æ³ã®å©ç¹ãšç¹åŸŽã«åºã¥ããŠãããã€ãã®ç 究ã°ã«ãŒãã¯ALKãã¹ ãã®èšºæã¢ã«ãŽãªãºã ãææ¡ããŠããïŒå³ 1-6ïŒïŒJapanese Lung Cancer Society 2011, Kim 2011, Paik 2011, Thunnissen 2012b, Conklin 2013, Marchetti 2013ïŒã Lung Cancer Samples
Cytology samples, frozen
RT-PCR Negative
IHC
Positive
Positive
Negative
FISH
Negative (rare)
Positive ALK inhibitor
å³ 1ãIHC ã«ããã¹ã¯ãªãŒãã³ã°ãš FISH ã« ãã確èªãããªã ãæ¥æ¬èºçåäŒãææ¡ãã ã¢ã«ãŽãªãºã ; RT-PCR ã¯çŽ°è蚺ã®æšæ¬ã«äœ¿çš ããããç¹ç·ã¯èšåºç çåŠçç¹åŸŽã«åŸã£ãŠæœè¡ å¯èœãªæ€æ»ãè¡šãã Biomarker Committee, Japanese Lung Cancer Societyã«ããèºçæ£ è ã®ALKéºäŒåæ€æ»ã®ããã®ã¬ã€ãã³ã¹ã第 1.0 ç 2011幎8æ1æ¥ãhttp://www haigan. gr.jp/uploads/photos/641.pdf ã§åç §å¯èœã
51
第 6 ç« : ALKãã¹ãã®ããã®ç°ãªã£ã解æãã©ããããŒã ã®æ¯èŒ
å³ 2ãIHC ããã³ CISH ãŸã㯠FISH ãçšããALKéºäŒååæ§æãäºæž¬ãã㢠ã«ãŽãªãºã ãKim H, Yoo S-B, Choe J-Y, et al ã®èš±å¯ãåŸãŠä¿®æ£ã J Thorac Oncol. 2011; 6 (8): 1359-1366ã
IHC
Negative 0/1+
Equivocal 2+
Positive 3+
Reported as ALK negative
CISH/FISH
Reported as ALK positive
å³ 3ãNSCLC ã®ALK IHC ãã ã³ FISH ã䜿çšãã蚺æã¢ã«ãŽãªãº ã ãPaik J, Choe G, Kim H, et al ã® èš±å¯ãåŸãŠä¿®æ£ã J Thorac Oncol. 2011;6(3):466-472.
IHC
Negative 0/1+
Equivocal 2+
Positive 3+
Reported as ALK negative
FISH
Reported as ALK positive
å³ 4ãALK IHC ãååã«ç¢ºèªãã ãå Žåã§ã® NSCLCALKãã¹ãã®ã ãã®ã¢ã«ãŽãªãºã ãThunnissen E, Bubendorf L, Dietel M, et al ã® èš±å¯ãåŸãŠä¿®æ£ã Virchows Arch. 2012;461(3):245-257.
NSCLC ALK testing IHC
Positive 1+, 2+, 3+
Negative
FISH
Negative
No ALK rearrangement
Positive
ALK rearrangement
52
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å³ 5ãIHC ã«ããALKãã¹ããš FISH ã« ãã確èªãããªã蚺æã¹ã¯ãªãŒãã³ã° ã®ã¢ã«ãŽãªãºã ãConklin C, Craddock KJ, Have C, et al ã®èš±å¯ãåŸãŠä¿®æ£ã J Thorac Oncol. 2013;8(1):45-51.
ALK IHC
(5A4 by Novocastra or DSF3 by Cell Signaling with ADVANCE detection)
Negative
Positive
(No ALK expression)
(Any ALK expression)
FISH ALK negative
ALK positive
NSCLC
(adenocarcinoma, large cell carcinoma, mixed tumors with adenocarcinoma, NOS)
ALK IHC
1-2 working days
Negative
Positive
FISH test
Negative No ALK rearrangement
ãŸãšã
2-3 working days
å³ 6ãthe Italian Association of Medical Oncology and the Italian Society of Pathology and Cytopathology ã«ãã NSCLCALKãã¹ãã®ã¢ã«ãŽãªãºã ãç¹ç· ã§æžãããããã¯ã¹ããã³ç¹ç·ã¯ãIHC ã«ããã¹ã¯ãªãŒãã³ã°ã®åŸã§ FISH ã«å ã仮説çãªæ¹æ³ã瀺ããŠãããIHC ãã ã³ FISHãã¹ããšçµæã®è§£éã«å¿ èŠãªæ éãèšèŒãããŠãããNOS=not otherwise specified. Marchetti A, Ardizzoni A, Papotti M et al ã®èš±å¯ãåŸãŠä¿®æ£ã J Thorac Oncol. 2013;8(3):352-358.
Positive ALK rearrangement
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NSCLC ã«ããã现èåŠã®åœ¹å²
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72
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Sakairi Y, Nakajima T, Yasufuku K, et al. EML4-ALK fusion gene assessment using metastatic lymph node samples obtained by endobronchial ultrasound-guided transbronchial needle aspiration. Clin Cancer Res. 2010;16(20):4938-4945. Salido M, Pijuan L, Martinez-Aviles L, et al. Increased ALK gene copy number and amplification are frequent in non-small cell lung cancer. J Thorac Oncol. 2011;6(1):21-27. Sanders H, Li HR, Bruey JM, et al. Exon scanning by reverse transcriptase-polymerase chain reaction for detection of known and novel EML4-ALK fusion variants in non-small cell lung cancer. Cancer Genet. 2011;204:45-52. Sasaki T, Rodig SJ, Chirieac LR, et al. The biology and treatment of EML4-ALK non-small cell lung cancer. Eur J Cancer. 2010;46:1773-1780. Savic S, Bode B, Diebold J, et al. Detection of ALK-positive non-small-cell lung cancers on cytological specimens: high accuracy of immunocytochemistry with the 5A4 clone. J Thorac Oncol. 2013;8(8):1004-1011. Saxe D, Persons DL, Wolff DJ, et al. Validation of fluorescence in situ hybridization using an analyte-specific reagent for detection of abnormalities involving the mixed lineage leukemia gene. Arch Pathol Lab Med. 2012;136(1):47-52. Schildhaus H, Deml KF, Schmitz K, et al. Chromogenic in situ hybridization is a reliable assay for detection of ALK rearrangements in adenocarcinomas of the lung. Mod Pathol. 2013 Jun 7 [Epub ahead of print]. Schreiber G, McCrory DC. Performance characteristics of different modalities for diagnosis of suspected lung cancer: summary of published evidence. Chest. 2003;123(1 Suppl):115S-128S. Selinger C, Rogers TM, Russell PA, et al. Testing for ALK rearrangement in lung adenocarcinoma: a multicenter comparison of immunohistochemistry and fluorescent in situ hybridization. Mol Pathol. 2013 Jun 7 [Epub ahead of print]. Shaozhang Z, Xiaomei L, Aiping Z, et al. Detection of EML4ALK fusion genes in non-small cell lung cancer patients with clinical features associated with EGFR mutations. Genes Chromsomes Cancer. 2012;51(10):925-932. Shaw A, Kim DW, Nakagawa K, et al. Crizotinib versus chemotherapy in advanced ALK-positive lung cancer. N Engl J Med. 2013;368(25):2385-2394. Shaw A, Yeap BY, Mino-Kenudson M, et al. Clinical features and outcome of patients with non-small-cell lung cancer who harbor EML4-ALK. J Clin Oncol. 2009;27(26):4247-4253. Shaw A, Yeap BY, Solomon BJ, et al. Effect of crizotinib on overall survival in patients with advanced non-small-cell lung cancer harbouring ALK gene rearrangement: a retrospective analysis. Lancet Oncol. 2011;12(11):1004-1012.
Soda M, Choi YL, Enomoto M, et al. Identification of the transforming EML4-ALK fusion gene in non-small-cell lung cancer. Nature. 2007;448(7153):561-566. Soda M, Isobe K, Inoue A, et al. A prospective PCR-based screening for the EML4-ALK oncogene in non-small cell lung cancer. Clin Cancer Res. 2012;18(20):5682-5689. Sun J, Choi YL, Won JK, et al. A dramatic response to crizotinib in a non-small-cell lung cancer patient with IHC-positive and FISH-negative ALK. J Thorac Oncol. 2012;7:e36-e38. Sun Y, Ren Y, Fang Z, et al. Lung adenocarcinoma from East Asian never-smokers is a disease largely defined by targetable oncogenic mutant kinases. J Clin Oncol. 2010;28(30):4616-4620. Takahashi T, Sonobe M, Kobayashi M, et al. Clinicopathologic features of non-small-cell lung cancer with EML4-ALK fusion gene. Ann Surg Oncol. 2010;17(3):889-897. Takamochi K, Takeuchi K, Hayashi T, et al. A rational diagnostic algorithm for the identification of ALK rearrangement in lung cancer: a comprehensive study of surgically treated Japanese patients. PLoS One. 2013;8(8):e69794. Takeuchi K. Interpretation of anti-ALK immunohistochemistry results [letter]. J Thorac Oncol. 2013;8(7):e67-e68. Takeuchi K, Choi YL, Soda M, et al. Multiplex reverse transcription-PCR screening for EML4-ALK fusion transcripts. Clin Cancer Res. 2008;14(20):6618-6624. Takeuchi K, Choi YL, Togashi Y, et al. KIF5B-ALK, a novel fusion oncokinase identified by an immunohistochemistry-based diagnostic system for ALK-positive lung cancer. Clin Cancer Res. 2009;15(9):3143-3149. Takeuchi K, Soda M, Togashi Y, et al. RET, ROS1 and ALK fusions in lung cancer. Nature Med. 2012;18:378-381. Tanaka H, Tone K, Hayashi A, et al. Clinical application of immunocytochemical detection of ALK rearrangement on cytology slides for detection or screening of lung adenocarcinoma. Lung Cancer. 2013;80(3):289-292. Thomas J, Lamb D, Ashcroft T, et al. How reliable is the diagnosis of lung cancer using small biopsy specimens? Report of a UKCCCR Lung Cancer Working Party. Thorax. 1993; 48(11):1135-1139. Thunnissen E, Boers E, Heideman DA, et al. Correlation of immunohistochemical staining p63 and TTF-1 with EGFR and K-ras mutational spectrum and diagnostic reproducibility in non small cell lung carcinoma. Virchows Arch. 2012;461(6):629-638. (a) Thunnissen E, Bubendorf L, Dietel M, et al. EML4-ALK testing in non-small cell carcinomas of the lung: a review with recommendations. Virchows Arch 2012;461(3):245-257. (b)
Shinmura K, Kageyama S, Tao H, et al. EML4-ALK fusion transcripts, but no NPM-, TPM3-, CLTC-, ATIC-, or TFG-ALK fusion transcripts, in non-small cell lung carcinomas. Lung Cancer. 2008;61(2):163-169.
Thunnissen E, Kerr KM, Bubendorf L, et al. External quality assessment for ALK immunohistochemistry testing in lung adenocarcinoma within the European Thoracic Oncology Platform Lungscape Project. Ann Oncol. 2012;23(suppl 9):2932. (c)
Sholl L, Weremowicz S, Gray SW, et al. Combined use of ALK immunohistochemistry and FISH for optimal detection of ALK-rearranged lung adenocarcinomas. J Thorac Oncol. 2013;8(3):322-328.
Thunnissen E, Kerr KM, Herth FJ, et al. The challenge of NSCLC diagnosis and predictive analysis on small samples. Practical approach of a working group. Lung Cancer. 2012;76(1):1-18. (d)
73
åèæç®
To KF, Tong JH, Yeung KS, et al. Detection of ALK rearrangement by immunohistochemistry in lung adenocarcinoma and the identification of a novel EML4-ALK variant. J Thorac Oncol. 2013;8(7):883-891.
Wong D, Leung EL, So KK, et al. The EML4-ALK fusion gene is involved in various histologic types of lung cancers from nonsmokers with wild-type EGFR and KRAS. Cancer. 2009;115(8):1723-1733.
Togashi Y, Soda M, Sakata S, et al. KLC1-ALK: a novel fusion in lung cancer identified using a formalin-fixed paraffinembedded tissue only. PLoS One. 2012;7:e31323.
Wu S, Kuo YY, Chang YL, et al. EML4-ALK translocation predicts better outcome in lung adenocarcinoma patients with wildtype EGFR. J Thorac Oncol. 2012;7(1):98-104.
Travis W, Brambilla E, Noguchi M, et al. International Association for the Study of Lung Cancer/American Thoracic Society/ European Respiratory Society international multidisciplinary classification of lung adenocarcinoma. J Thorac Oncol. 2011; 6(2):244-285.
Yang P, Kulig K, Boland JM, et al. Worse disease-free survival in never-smokers with ALK+ lung adenocarcinoma. J Thorac Oncol. 2012;7(1):90-97.
Travis W, Brambilla E, Noguchi M, et al. Diagnosis of lung cancer in small biopsies and cytology: implications of the 2011 International Association for the Study of Lung Cancer/ American Thoracic Society/European Respiratory Society classification. Arch Pathol Lab Med. 2013;137(5):668-684. Tsao M, Craddock K, Brandao G, et al. Canadian ALK (CALK): A pan-Canadian multicenter study to optimize and standardize ALK immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) for ALK gene rearrangements. J Clin Oncol. 2013;31(May 20 Supplement):8096. V Laffert M, Warth A, Penzel R, et al. Anaplastic lymphoma kinase (ALK) gene rearrangement in non-small cell lung cancer (NSCLC): results of a multi-centre ALK-testing. Lung Cancer. 2013;81:200-206. van Krieken J, Normanno N, Blackhall F, et al. Guideline on the requirements of external quality assessment programs in molecular pathology. Virchows Arch. 2013;462:27-37. Wang R, Hu H, Pan Y, et al. RET fusions define a unique molecular and clinicopathologic subtype of non-small-cell lung cancer. J Clin Oncol. 2012;30:4352-4359. (a) Wang R, Pan Y, Li C, et al. The use of quantitative real-time reverse transcriptase PCR for 5â and 3â portions of ALK transcripts to detect ALK rearrangements in lung cancers. Clin Cancer Res. 2012;18(4725-4732. (b)
Yi E, Boland JM, Maleszewski JJ, et al. Correlation of IHC and FISH for ALK gene rearrangement in non-small cell lung carcinoma: IHC score algorithm for FISH. J Thorac Oncol 2011;6:459-465. Yi E, Chung JH, Kulig K, Kerr KM. Detection of anaplastic lymphoma kinase (ALK) gene rearrangement in non-small cell lung cancer and related issues in ALK inhibitor therapy: a literature review. Mol Diagn Ther. 2012;16:143-150. Yoshida A, Tsuta K, Nitta H, et al. Bright-field dual-color chromogenic in situ hybridization for diagnosing echinoderm microtubule-associated protein-like 4-anaplastic lymphoma kinase-positive lung adenocarcinomas. J Thorac Oncol. 2011;6(10):1677-1686. (a) Yoshida A, Tsuta K, Watanabe S, et al. Frequent ALK rearrangement and TTF-1/p63 co-expression in lung adenocarcinoma with signet-ring cell component. Lung Cancer. 2011;72:309315. (b) Zhang X, Zhang S, Yang X, et al. Fusion of EML4 and ALK is associated with development of lung adenocarcinomas lacking EGFR and KRAS mutations and is correlated with ALK expression. Mol Cancer. 2010;9:188. Zhang Y-G, Jin M-L, Li L, et al. Evaluation of ALK rearrangement in Chinese non-small cell lung cancer using FISH, immunohistochemistry, and real-time quantitative RT-PCR on paraffin-embedded tissues. PLOS One. 2013;8(5):e64821.
Wang Z, Zhang X, Bai H, et al. EML4-ALK rearrangement and its clinical significance in Chinese patients with advanced non-small cell lung cancer. Oncology. 2012;83(5):248-256.
Zhang Y, Sun Y, Pan Y, et al. Frequency of driver mutations in lung adenocarcinoma from female never-smokers varies with histologic subtypes and age at diagnosis. Clin Cancer Res. 2012;18(7):1947-1953.
Werner M, Chott A, Fabiano A, Battifora H. Effect of formalin tissue fixation and processing on immunohistochemistry. Am J Surg Pathol. 2000;24(7):1016-1019.
Zhou J, Yang H, Deng Q, et al. Oncogenic driver mutations in patients with non-small-cell lung cancer at various clinical stages. Ann Oncol. 2013;24(5):1319-1325.
Wolff A, Hammond ME, Schwartz JN, et al. American Society of Clinical Oncology/College of American Pathologists guideline recommendations for human epidermal growth factor receptor 2 testing in breast cancer. Arch Pathol Lab Med. 2007;131(1):18-43.
ç±³åœ/éåœ
ç±³åœ/ã¹ã€ã¹ FISH/RT-PCR
æ¥æ¬
æ¥æ¬
ç±³åœ
Koivunen et al., 2008
Perner et al., 2008
Shinmura et al., 2008
Takeuchi et al., 2008
Boland et al., 2009
IHC/FISH
ç±³åœ
ç±³åœ
æ¥æ¬
äžåœ(HK)
æ¥æ¬
æ¥æ¬
ã¹ãã€ã³
äžåœ
æ¥æ¬
Rodig et al., 2009
Shaw et al., 2009
Takeuchi et al., 2009
Wong et al., 2009
Jokoji et al., 2010
Sakairi et al., 2010
Salido et al., 2010
Sun Y et al., 2010
Takahashi et al., 2010
IHC/FISH
éåœ
ç±³åœ
Paik et al., 2011
Yi et al., 2011
FISH/IHC
RACE/ RT-PCR
Zhang X et al., 2010 äžåœ
RT-PCR/Direct Seq
RT-PCR
FISH/IHC
IHC/FISH & RT-PCR
IHC
RT-PCR
IHC/RT-PCR & FISH
FISH
RT-PCR/FISH/IHC
ã€ã¿ãªã¢ïŒ Martelli et al., 2009 ã¹ãã€ã³
IHC/FISH/RTPCR
RT-PCR/FISH/ Gen Seq
RT-PCR/Gen Seq
RT-PCR
RT-PCR
æ¥æ¬
RT-PCR
äžåœ
æ€æ»æ¹æ³
Soda et al., 2007
åœ
Rikova et al., 2007
ç 究
101
735
103
313
52
107
109
254
266
130
141
358
120
335
340
77
603
305
75
103
NSCLC* æ°
50
-
208
20
-
ADC æ°
63
185
130
62
12 (11.7%)
10 (9.9%) 101
28 (3.8%) 395
211
52
69
82
254
5 (1.6%)
3 (5.8%)
3 (2.8%)
7 (6.4%)
8 (3.1%)
20 (5.6%)
3 (4.8%)
5 (2.7%)
11 (4.3%)
2 (4.0%)
-
8 (3.8%)
2 (10.0%)
-
ALK+ æ° (%)
10 (9.9%)
27(6.8%)
10 (16.1%)
5 (2.4%)
3 (5.8%)
3 (4.3%)
7 (8.5%)
8 (3.1%)
11 (5.3%)
4 (3.1%)
130 18 (12.8%)
13 (4.9%) 209
4 (3.1%)
19 (13.5%)
20 (5.6%) 358
9 (7.5%)
6 (1.8%)
11 (3.2%) 253
2 (2.6%)
16 (2.7%)
8 (2.6%)
5 (6.7%)
4 (4%)
ALK+ æ° (%)
SCC æ°
-
-
0
0
-
0
1 (9.1%)
0
292
29
75
0
30
18
0
34
0
2
0
48
0
0
2 (6.9%)
0
0
0
0
0
0
0
0
0
4 (8.3%)
150 1 (0.7%)
71
20
-
88
11
0
0
0
0
0
0
0
0
0
0
4
0
2
0
7
3
-
9
2
-
0
0
0
0
0
0
0
0
0
0
1 (25%)
0
2 (100%)
0
0
0
-
0
0
-
0
48
12
27
0
8
9
0
23
0
5
0
7
0
9
4
-
0
42
0
1 (2.1%)
0
0
0
0
0
0
2 (8.7%)
0
0
0
0
0
0
0
-
0
0
-
0
0
0
0
0
0
0
0
0
0
0
0
0
0
24
0
-
0
0
-
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
-
0
0
-
ALK+ ADSC ALK+ NSCLCã ALK+ NECa ALK+ æ° (%) æ° æ° (%) ä»æ° æ° (%) æ° æ° (%)
å ±åãããèºçã«ãããALKéºäŒååæ§ææ€æ»ã«é¢ããç 究ã®æŠèŠ
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SCLC LCNEC ãªã©ã®ç¥çµå åæ³çãå«ã
æ£è ã® éžæ
èºçã«ãããALKãã¹ãIASLC ã¢ãã©ã¹
74
ç±³åœ
éåœ
Dai et al., 2012
Jin et al., 2012
äžåœ
ã¹ãã€ã³
ç±³åœ
ç±³åœ
äžåœ(HK)
äžåœ
äžåœ
Li Y et al., 2013
Martinez et al., 2013
Minca et al., 2013
Sholl et al., 2013
To et al., 2013
Zhang Y-G et al., 2013
Zhou et al., 2013
ARMS RT-PCR
RT-PCR/IHC/FISH
IHC/FISH/RT-PCR
IHC/FISH
IHC/FISH
FISH/IHC
12665
488
469
373
186
249
99
208
1683
349
300
116
113
102
258
811
441
167
1387
239 -
-
79
95
-
662 (5.2%)
6775
28 (5.7%) 349
20 (4.2%) 341
22 (5.9%) 373
12 (6.5%) 186
32(12.8%)
7 (7.1%)
7 (3.4%)
75 (4.4%)
15 (4.3%) 349
444 (6.6%)
25 (7.2%)
19 (2.6%)
22 (5.9%)
12 (6.5%)
-
5 (6.3%)
6 (6.3%)
-
15 (4.3%)
0
0
-
0
0
-
0
0
0
1 (5.6%)
1 (7.1%)
0
1 (1.9%)
0
1 (0.2%)
-
4 (4.1%)
18 1411 (1.3%)
101 2 (2.0%)
112
0
-
7
96
-
0
0
18
14
34â(11.3%) 300 34 (11.3%)
10 (10.5%)
5 (6.2%)
0
52
0
46
-
97
0
116
95
73
17 (9.3%)
30 (4.2%)
29 (6.6%)
9 (7.4%)
-
10 (7.7%)
39 (34%)
39 (34%)
11 (9.7%)
8(7.8%)
25 (9.7%) 182
31 (3.8%) 713
29 (6.6%) 441
10 (6.0%) 121
49 (3.7%)
15 (6.3%) 130
78
18
4
0
-
0
7
-
0
0
0
0
14
0
8
0
0
-
-
7 (9.0%)
1(5.6%)
0
0
-
376
20
12
0
-
13
10 0
-
-
0
0
0
0
1
76
38
0
0
-
12
1 (14.3%)
0
0
0
0
2 (14.3%)
0
0
0
0
-
-
17 (4.5%)
0
1 (8.3%)
0
-
2 (15.4%)
0
-
0
0
0
0
28
0
0
0
-
0
0
-
0
0
0
0
0
4
8 (10.5%) 0
0
0
0
-
0
0
0
0
-
1 (8.3%)
0
0
0
0
-
0
0
-
0
0
0
0
0
0
0
0
0
-
0
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Total
FISH
Uç±³åœ
Gainor et al., 2013
RT-PCR/IHC
IHC/FISH
RACE-PCR
ç±³åœ
Yang et al., 2012
Zhang Y et al., 2012 äžåœ
RT-PCR/FISH
å°æ¹Ÿ
RT-PCR
Wu et al., 2012
äžåœ
Shaozhang et al., 2012
IHC/FISH
FISH/IHC
éåœ
Park et al., 2012
RT-PCR/IHC/FISH
IHC/FISH
RT-PCR
FISH
RACE-PCR
Wang Z et al., 2012 äžåœ
æ¥æ¬
Murakami et al., 2012
McLeer-Florin et al., ãã©ã³ã¹ 2012
äžåœ
An et al., 2012
ä»èš 1
75
76
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Section I .èºçéºäŒåæ€æ»ããã€è¡ãã¹ããïŒ
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5.1å°é家統äžèŠè§£ïŒ ç çå»ã¯ãEGFRãã¹ãã®ããã®æ€äœã®é©æ£ããç现èã®å²åãDNAã®è³ªããã³é ã«ããšã¥ããŠæ±ºå®ãã¹ãã§ããã 5.2å°é家統äžèŠè§£ïŒ ããããã®æ€æ»ã©ãã§ã¯ãå€äœæ€åºã®éã®æäœå«æéããã³çŽ°èæ°ãæ€èšŒå®éšã®é ã«æ±ºå®ããå¿ èŠãããã 5.3å°é家統äžèŠè§£ïŒ ç çå»ã¯ãããããã®æ€äœã«ãããè «çæåãåãåºããããã«ãã¹ãã§ãå¿ èŠã« å¿ããŠç çå»èªãããããã¯ããæå°ãããæåž«ã«ãã£ãŠè «çã«å¯ãã éšåãã〠ã¯ããã€ã»ã¯ããã¹ãã§ããã
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10.1aæšå¥šïŒ èºè ºçã«ãããŠã¯ãEGFRãã¹ãã¯ä»ã®ååããŒã«ãŒãããåªå çã«æ±ããã ã¹ãã§ããã 10.1bæèšïŒ EGFRãã¹ãã®æ¬¡ã«ãALKãã¹ãã¯èºè ºçã§ã¯åªå çã«æ±ãããã°ãªããªããã çŸç¶ã§ã¯ååãªãšããã³ã¹ã¯çºè¡šãããŠããªãã
Section V: èºè ºçã®éºäŒåãã¹ãã¯ããã«å®æœãããéçšãããã¹ããïŒ è«ç¹11 ãã¹ãŠã®è ºçããEGFRãšALKã®äž¡æ¹ã®ãã¹ãããããã¹ãã§ãããïŒ
11.1å°é家統äžèŠè§£: æ€æ»æœèšã¯ãå šäœã®çµæã¬ããŒãè¿åŽæéèŠä»¶ãæºããã€ã€ãè ºçã®éºäŒåã ã¹ãã®å¹çãæ倧åããæ€æ»ã¢ã«ãŽãªãºã ãå®æœãããšæãããã
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12.1å°é家統äžèŠè§£ïŒ EGFRå€ç°ãã¹ããšALK FISHã®å ±åæžã«ã¯ãè «çå ç§å»ãéå°é家ç çå»ã«ã 容æã«ç解ã§ããçµæãšè§£éã®é ç®ãå«ãŸããŠããå¿ èŠãããã
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13.1å°é家統äžèŠè§£: EGFRãšALKãã¹ãã®æå¹æ§ã®å®èšŒã«ã€ããŠã¯ãä»ã®éºäŒå蚺æãFISHãã¹ã ãšåæ§ã®ã¬ã€ãã©ã€ã³ãçšããã¹ãã§ããã
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14.1å°é家統äžèŠè§£: æ€æ»æœèšã¯ãèºçã®EGFRãALKãã¹ããä»ã®èšåºæ€æ»ã§çšãããã粟床管çã ãªã·ãŒã«åŸãã¹ãã§ãããç¹ã«ãæ²»çã®ããã®EGFR,ALKãã¹ããè¡ãæ€æ»æ ã¯ãæèœèª¿æ»ã«åå ãã¹ãã§ããã Abbreviations: CISH, chromogenic in situ hybridization; EGFR, epidermal growth factor receptor; FISH, fluorescence in situ hybridization; PCR, polymerase chain reaction; RT-PCR, reverse transcription-polymerase chain reaction; TKI, tyrosine kinase inhibitor; TNM, tumor node metastasis. 次ã®è«æããèš±å¯ãããŠè»¢èŒ: Lindeman NI, Cagle PT, Beasley MB, et al. Molecular testing guideline for selection of lung cancer patients for EGFR and ALK tyrosine kinase inhibitors: Guideline from the College of American Pathologists (CAP), International Association for the Study of Lung Cancer (IASLC), Association for Molecular Pathology (AMP). J Thorac Oncol 2013;8:823-859.
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