Author Manuscript Published OnlineFirst on November 3, 2011; DOI: 10.1158/0008-5472.CAN-11-2023
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ͳ
5-hydroxymethylcytosine is strongly depleted in human cancers
but its levels do not correlate with IDH1 mutations
‡—‰Ǧ
‹ ‹ǡͳǡʹ ‘‰ ‹ƒ‰ǡͳǡʹ —š‹ƒ‰ ‹—ǡͳǡ͵ ‹„‘” Ǥ ƒ—…ŠǡͶ ‹•Š‡‰ ƒ‰ǡͷ
ƒ„”‹‡Ž‡…Šƒ…‡”–ǡ͸‹‡–ƒ””‡šǡ͸‹ƒ‰—ǡ͵ƒ†
‡”†Ǥˆ‡‹ˆ‡”ʹȗ
ͳŠ‡•‡ƒ—–Š‘”•…‘–”‹„—–‡†‡“—ƒŽŽ›Ǥ
2Department
of Cancer Biology and 3Department of Neurosciences, Beckman Research
Institute of the City of Hope, Duarte, CA 91010, USA, 4Section of Molecular Medicine,
Department of Orthopedic Surgery, Rush University Medical Center, Chicago, IL 60612,
USA; 5Department of Chemistry, University of California Riverside, Riverside, CA 92521,
6Klinik
und Poliklinik für Neurochirurgie, Universitätsklinikum Carl Gustav Carus,
Technische Universität Dresden, 01307 Dresden, Germany.
* Correspondence:
”Ǥ
‡”†Ǥˆ‡‹ˆ‡”
‡’ƒ”–‡–‘ˆƒ…‡”‹‘Ž‘‰›
‡…ƒ‡•‡ƒ”…Š•–‹–—–‡
‹–›‘ˆ‘’‡
ͳͷͲͲǤ—ƒ”–‡‘ƒ†
—ƒ”–‡ǡͻͳͲͳͲ
’Š‘‡ǣ͸ʹ͸Ǧ͵ͲͳǦͺͺͷ͵
‡ƒ‹Žǣ‰’ˆ‡‹ˆ‡”̷…‘ŠǤ‘”‰
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 2011 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on November 3, 2011; DOI: 10.1158/0008-5472.CAN-11-2023
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ʹ
Abstract
Š‡„ƒ•‡ͷǦŠ›†”‘š›‡–Š›Ž…›–‘•‹‡ȋͷŠȌ™ƒ•”‡…‡–Ž›‹†‡–‹ˆ‹‡†ƒ•ƒ‘š‹†ƒ–‹‘
’”‘†—…– ‘ˆ ͷǦ‡–Š›Ž…›–‘•‹‡ ȋͷȌ ‹ ƒƒŽ‹ƒ Ǥ ‡”‡ǡ —•‹‰ •‡•‹–‹˜‡ ƒ†
“—ƒ–‹–ƒ–‹˜‡‡–Š‘†•–‘ƒ••‡••Ž‡˜‡Ž•‘ˆͷǦŠ›†”‘š›‡–Š›ŽǦʹ̵Ǧ†‡‘š›…›–‹†‹‡ȋͷŠ†Ȍ
ƒ† ͷǦ‡–Š›ŽǦʹ̵Ǧ†‡‘š›…›–‹†‹‡ ȋͷ†Ȍ ‹ ‰‡‘‹… ǡ ™‡ ‹˜‡•–‹‰ƒ–‡† ™Š‡–Š‡”
Ž‡˜‡Ž• ‘ˆ ͷŠ …ƒ †‹•–‹‰—‹•Š ‘”ƒŽ –‹••—‡ ˆ”‘ –—‘” –‹••—‡Ǥ  •“—ƒ‘—• …‡ŽŽ
Ž—‰ …ƒ…‡”•ǡ Ž‡˜‡Ž• ‘ˆ ͷŠ† ™‡”‡ †‡’Ž‡–‡† •—„•–ƒ–‹ƒŽŽ› ™‹–Š —’ –‘ ͷǦˆ‘Ž†
”‡†—…–‹‘…‘’ƒ”‡†–‘‘”ƒŽŽ—‰–‹••—‡Ǥ„”ƒ‹–—‘”•ǡͷŠ†•Š‘™‡†ƒ‡˜‡
‘”‡ †”ƒ•–‹… ”‡†—…–‹‘ ™‹–Š Ž‡˜‡Ž• —’ –‘ ε͵ͲǦˆ‘Ž† Ž‘™‡” –Šƒ ‹ ‘”ƒŽ „”ƒ‹ǡ „—–
ͷŠ†Ž‡˜‡Ž•™‡”‡‹†‡’‡†‡–‘ˆ—–ƒ–‹‘•‹‹•‘…‹–”ƒ–‡†‡Š›†”‘‰‡ƒ•‡ǦͳȋͳȌǤ
—”–Š‡”‘”‡ǡ ‹—‘Š‹•–‘…Š‡‹…ƒŽ ƒƒŽ›•‹• ‹†‹…ƒ–‡† –Šƒ– ͷŠ ‹• ”‡ƒ”ƒ„Ž›
†‡’Ž‡–‡† ‹ ƒ› –›’‡• ‘ˆ Š—ƒ …ƒ…‡”Ǥ ’‘”–ƒ–Ž›ǡ ƒ ‹˜‡”•‡ ”‡Žƒ–‹‘•Š‹’
„‡–™‡‡ ͷŠ Ž‡˜‡Ž• ƒ† …‡ŽŽ ’”‘Ž‹ˆ‡”ƒ–‹‘ ™ƒ• ‘„•‡”˜‡† ™‹–Š Žƒ… ‘ˆ ͷŠ ‹
’”‘Ž‹ˆ‡”ƒ–‹‰ …‡ŽŽ•Ǥ Š‡ †ƒ–ƒ –Š‡”‡ˆ‘”‡ •—‰‰‡•– –Šƒ– ͷŠ† ‹• •–”‘‰Ž› †‡’Ž‡–‡† ‹
Š—ƒ ƒŽ‹‰ƒ– –—‘”•ǡ ƒ ˆ‹†‹‰ –Šƒ– ƒ††• ƒ‘–Š‡” Žƒ›‡” ‘ˆ …‘’Ž‡š‹–› –‘ –Š‡
ƒ„‡””ƒ–‡’‹‰‡‘‡ˆ‘—†‹…ƒ…‡”–‹••—‡Ǥƒ††‹–‹‘ǡƒŽƒ…‘ˆͷŠƒ›„‡…‘‡
ƒ—•‡ˆ—Ž„‹‘ƒ”‡”ˆ‘”…ƒ…‡”†‹ƒ‰‘•‹•Ǥ
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 2011 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on November 3, 2011; DOI: 10.1158/0008-5472.CAN-11-2023
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
͵
Introduction
‡…‡–Ž›ǡͷŠŠƒ•„‡‡‹†‡–‹ˆ‹‡† ƒ•ƒ‘š‹†ƒ–‹‘’”‘†—…–‘ˆͷ‹ƒƒŽ‹ƒ
ȋͳǡʹȌ‰‡‡”ƒ–‡†„›–Š‡ƒŽ’ŠƒǦ‡–‘‰Ž—–ƒ”ƒ–‡Ǧ†‡’‡†‡–‡–†‹‘š›‰‡ƒ•‡•ȋʹǡ͵ȌǤ
‡˜‡Ž•‘ˆͷŠƒ”‡–‹••—‡Ǧ†‡’‡†‡–ƒ†–Š‡Š‹‰Š‡•–Ž‡˜‡Ž•Šƒ˜‡„‡‡ˆ‘—†‹–Š‡
…‡–”ƒŽ ‡”˜‘—• •›•–‡ ȋͶǡ ͷȌǤ Š‡ „‹‘Ž‘‰‹…ƒŽ ˆ—…–‹‘ ‘ˆ ͷŠ ‹• …—””‡–Ž›
—‘™Ǥ ͷŠ ƒ› „‡ ƒ ‹–‡”‡†‹ƒ–‡ ‹ †‡‡–Š›Žƒ–‹‘ ’”‘…‡••‡• –Šƒ–
ƒ……‘’Ž‹•Š‡• –Š‡ …‘˜‡”•‹‘ ‘ˆ ͷ –‘ …›–‘•‹‡ ȋ͸ȌǤ Š‡ †‹•–”‹„—–‹‘ ‘ˆ ͷŠ ‹
‡„”›‘‹…•–‡…‡ŽŽ•ȋ͹ǦͳʹȌǡ‘—•‡…‡”‡„‡ŽŽ—ȋͳ͵Ȍǡƒ†Š—ƒ’”‡ˆ”‘–ƒŽ…‘”–‡š
ȋͳͶȌŠƒ•„‡‡ƒ’’‡†„›ƒ””ƒ›Ǧ‘”•‡“—‡…‹‰Ǧ„ƒ•‡†ƒ••ƒ›•ǤͷŠ™ƒ•‡”‹…Š‡†ƒ–
’”‘‘–‡”•ƒ†™‹–Š‹‰‡‡„‘†‹‡•Ǥ
‡˜‡”ƒŽŠ‡ƒ–‘Ž‘‰‹…ƒŽƒŽ‹‰ƒ…‹‡•…ƒ””›—–ƒ–‹‘•‹‘‡‘ˆ–Š‡TET‰‡‡•ǡ
TET2ȋͳͷȌǤTET2—–ƒ–‹‘•™‡”‡Ž‹‡†–‘ƒ„‡””ƒ–Ž‡˜‡Ž•‘ˆͷŠƒ†ͷ‹–Š‡•‡
…ƒ…‡” ‰‡‘‡• ȋͳ͸ǡ ͳ͹ȌǤ Ž•‘ǡ —–ƒ–‹‘• ‹ ͳ Šƒ˜‡ „‡‡ Ž‹‡† –‘ ƒ„‘”ƒŽ
‡–Š›Žƒ–‹‘ ’ƒ––‡”• ȋͳͺȌǤ ‡ ƒ––”ƒ…–‹˜‡ ’”‘’‘•ƒŽ ‹• –Šƒ– —–ƒ–‡† ͳ
’”‘†—…‡• ƒ ‡™ ‡–ƒ„‘Ž‹–‡ǡ ʹǦŠ›†”‘š›‰Ž—–ƒ”ƒ–‡ ȋʹ
Ȍ ȋͳͻȌǡ ™Š‹…Š …ƒ ‹Š‹„‹– ’”‘–‡‹•’‘–‡–‹ƒŽŽ›Ž‡ƒ†‹‰–‘ƒŽ–‡”‡†Ž‡˜‡Ž•‘ˆͷŠƒ†ͷ‹–—‘”•ȋʹͲȌǤ
›•–‡ƒ–‹…•–—†‹‡•‘Ž‡˜‡Ž•‘ˆͷŠ‹Š—ƒ…ƒ…‡”•ƒ”‡Žƒ…‹‰Ǥ‡”‡ǡ™‡
Šƒ˜‡—•‡†Ž‹“—‹†…Š”‘ƒ–‘‰”ƒ’Š›–ƒ†‡ƒ•••’‡…–”‘‡–”›ȋǦȀȌǡ–‘ƒ••‡••
–Š‡Ž‡˜‡Ž•‘ˆͷŠ†ƒ†ͷ†‹Š—ƒŽ—‰…ƒ”…‹‘ƒ•ƒ†‹„”ƒ‹–—‘”Ǥ
‡ƒŽ•‘—•‡†‹—‘ˆŽ—‘”‡•…‡…‡•–ƒ‹‹‰–‘ƒ••‡••ͷŠ‹ƒ•‡”‹‡•‘ˆ‘”ƒŽƒ†
ƒŽ‹‰ƒ––‹••—‡•‡…–‹‘•Ǥ
Materials and Methods
DNA samples:
–ƒ‰‡Ǧ Ž—‰ •“—ƒ‘—• …‡ŽŽ …ƒ”…‹‘ƒ ȋȌ ƒ† ƒ†‡‘…ƒ”…‹‘ƒ •ƒ’Ž‡• ƒ†
ƒ–…Š‡† ‘”ƒŽ –‹••—‡• ™‡”‡ ‘„–ƒ‹‡† ˆ”‘ –Š‡ ˆ”‘œ‡ –—‘” „ƒ ‘ˆ –Š‡ ‹–› ‘ˆ
‘’‡ ‡†‹…ƒŽ ‡–‡” —†‡” ƒ •–‹–—–‹‘ƒŽ ‡˜‹‡™ ‘ƒ”† ƒ’’”‘˜‡† ’”‘–‘…‘ŽǤ
ƒ’Ž‡• ™‡”‡ ‘„–ƒ‹‡† ˆ”‘ –—‘”• ™‹–Š‘—– Žƒ•‡”Ǧ…ƒ’–—”‡ ‹…”‘†‹••‡…–‹‘Ǥ ˆ”‘’”‹ƒ”›•ƒŽŽ…‡ŽŽŽ—‰…ƒ…‡”•ƒ†ƒ–…Š‡†‘”ƒŽŽ—‰™ƒ•‘„–ƒ‹‡†ˆ”‘
•–‡”ƒ†ȋ‡–”‘‹–ǡȌǡ‹‘Šƒ‹ȋƒ›™ƒ”†ǡȌǡƒ†—”‡Ž‹‡ȋ‘—–Šƒ”ƒ…‹•…‘ǡ
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 2011 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on November 3, 2011; DOI: 10.1158/0008-5472.CAN-11-2023
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Ͷ
Ȍ. ‘”ƒŽŠ—ƒ„”ƒ‹–‹••—‡•‘ˆ–Š‡’”‡ˆ”‘–ƒŽ…‘”–‡š™‡”‡‘„–ƒ‹‡†ˆ”‘
ƒ’‹–ƒŽ ‹‘•…‹‡…‡• ȋ
ƒ‹–Š‡”•„—”‰ǡ Ȍ ƒ† ‹‘Šƒ‹ ȋƒ›™ƒ”†ǡ ȌǤ ˆ”‘
‡—”‘•ƒ†ƒ•–”‘…›–‡•‘ˆˆ‡–ƒŽȋʹͶ™‡‡•‘ˆ‰‡•–ƒ–‹‘ȌŠ—ƒ„”ƒ‹™ƒ•‘„–ƒ‹‡†
ˆ”‘ …‹‡‡ŽŽ ȋƒ”Ž•„ƒ†ǡ ȌǤ ™‡–›Ǧ•‡˜‡ ƒ•–”‘…›–‘ƒ• ȋ‘”Ž† ‡ƒŽ–Š
”‰ƒ‹œƒ–‹‘ ‰”ƒ†‡ ǦȌ ™‡”‡ ‘„–ƒ‹‡† ‘ •–‹–—–‹‘ƒŽ ‡˜‹‡™ ‘ƒ”† ƒ’’”‘˜‡†
’”‘–‘…‘Ž•ƒ––Š‡‡’ƒ”–‡–‘ˆ‡—”‘•—”‰‡”›ƒ––Š‡‹˜‡”•‹–›‘•’‹–ƒŽ‹”‡•†‡Ǥ
™ƒ•‹•‘Žƒ–‡†„›•–ƒ†ƒ”†’”‘…‡†—”‡•—•‹‰’Š‡‘ŽǦ…ŠŽ‘”‘ˆ‘”‡š–”ƒ…–‹‘ƒ†
‡–Šƒ‘Ž ’”‡…‹’‹–ƒ–‹‘Ǥ ‹‰Š– ƒ††‹–‹‘ƒŽ „”ƒ‹ –—‘” • ™‡”‡ ‘„–ƒ‹‡† ˆ”‘
•–‡”ƒ† ȋ‡–”‘‹–ǡ ȌǤ ‡‘‹… • ˆ”‘ –‹••—‡• ƒ† …‡ŽŽ Ž‹‡• ™‡”‡ ‹•‘Žƒ–‡†
—•‹‰–Š‡‡ƒ•›‹••—‡‹–ȋ‹ƒ‰‡ǢƒŽ‡…‹ƒǡȌǤ
IDH mutations:
‘”•‡“—‡…‹‰‘ˆIDH1ƒ†IDH2‡š‘ͶǡͶͲ‰‘ˆ‰‡‘‹…™ƒ•—•‡†ˆ‘”
ƒ’Ž‹ˆ‹…ƒ–‹‘ —•‹‰ –Š‡ ˆ‘ŽŽ‘™‹‰ ’”‹‡”•ǣ ˆ‘” IDH1ǡ –Š‡ ˆ‘”™ƒ”† ’”‹‡” ͷǯǦ
Ǣ
ƒ†
–Š‡
ˆ‘”
IDH2ǡ
”‡˜‡”•‡
–Š‡
ˆ‘”™ƒ”†
’”‹‡”
’”‹‡”
ͷǯǦ
ͷǯǦ
ƒ†–Š‡”‡˜‡”•‡’”‹‡”ͷǯǦ
Ǥ
Simultaneous quantification of 5mdC and 5hmdC by LC-MS/MS
‡‘‹…ȋͳ–‘ʹρ‰Ȍ™ƒ•‹…—„ƒ–‡†™‹–Šͷ—‹–•‘ˆ‡‰”ƒ†ƒ•‡Ž—•ȋ›‘
‡•‡ƒ”…ŠǢ”˜‹‡ǡȌƒ–͵͹ιˆ‘”ƒ–Ž‡ƒ•–ʹŠ‘—”•ǤŠ‡•–ƒ„Ž‡‹•‘–‘’‡Žƒ„‡Ž‡†ͷŠ†
ȋʹͳǡ ʹʹȌǡ ƒ† Žƒ„‡Ž‡† ʹǯǦ†‡‘š›‰—ƒ‘•‹‡ ȋƒ„”‹†‰‡ •‘–‘’‡ ƒ„‘”ƒ–‘”‹‡•Ǣ
ƒ„”‹†‰‡ǡ Ȍ ™‡”‡ ƒ††‡† ƒ• ‹–‡”ƒŽ •–ƒ†ƒ”†•Ǥ Ž‹“—‘–• ‘ˆ –Š‡ ‹š–—”‡ ™‡”‡
•—„Œ‡…–‡††‹”‡…–Ž›–‘ǦȀƒƒŽ›•‹•Ǥ ǦȀ™ƒ•’‡”ˆ‘”‡†—•‹‰ƒŠ‡”‘
……‡Žƒ͸ͲͲ’—’‹–‡”ˆƒ…‡†™‹–Šƒƒ–ƒ‰‡–”‹’Ž‡•–ƒ‰‡“—ƒ†”—’Ž‡ƒ••
•’‡…–”‘‡–‡”ȋŠ‡”‘‹•Š‡”…‹‡–‹ˆ‹…ȌǤʹǤͳšͷͲ‹‡–‡šǦͳͺ…‘Ž—ȋʹǤ͸
Ɋ‹’ƒ”–‹…Ž‡•‹œ‡ƒ†ͳͲͲ%‹’‘”‡•‹œ‡ǢŠ‡‘‡‡šǢ‘””ƒ…‡ǡȌ™ƒ•—•‡†ˆ‘”
•‡’ƒ”ƒ–‹‘ƒ–ƒˆŽ‘™”ƒ–‡‘ˆͶͲͲμŽȀ‹ǤŠ‡ƒ•••’‡…–”‘‡–‡”™ƒ•‘’–‹‹œ‡†
ƒ†•‡–—’‹•…ƒ‘†‡ˆ‘”‘‹–‘”‹‰–Š‡ȏΪȐΪ‹‘•‘ˆͷŠ†ȋm/zʹͷͺǤͳ
ൺͳͶʹǤͳȌǡͷ†ȋm/zʹͶʹǤͳൺͳʹ͸ǤͳȌǡ†
ȋm/zʹ͸ͺǤͳൺͳͷʹǤͳȌǡŽƒ„‡Ž‡†ͷŠ†ȋm/z
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Author Manuscript Published OnlineFirst on November 3, 2011; DOI: 10.1158/0008-5472.CAN-11-2023
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ͷ
ʹ͸ͳǤͳ ൺ ͳͶͶǤͳȌ ƒ† Žƒ„‡Ž‡† †
ȋm/z ʹ͹͵Ǥͳ ൺ ͳͷ͹ǤͳȌǤ Š‡”‘ …ƒŽ‹„—” •‘ˆ–™ƒ”‡
˜‡”•‹‘ʹǤͳ™ƒ•—•‡†–‘’‡”ˆ‘”†ƒ–ƒƒƒŽ›•‹•Ǥ
—‘†‘–„Ž‘–ƒƒŽ›•‹•ˆ‘”ͷŠ™ƒ•’‡”ˆ‘”‡†ƒ•†‡•…”‹„‡†’”‡˜‹‘—•Ž›
ȋͳͶȌǤ
Immunohistochemistry
”‘œ‡ –‹••—‡ ƒ””ƒ›• ™‡”‡ ˆ”‘ ‹‘…Šƒ‹ ȋ…ƒ–ƒŽ‘‰ ‘Ǥ ͸ʹ͵ͷ͹ͲͲǦͷǢ Ž‘– ‘Ǥ
ͶͲ͵ͳͲͻȌǤ Š‡› …‘–ƒ‹ ‘”ƒŽ „”ƒ‹ –‹••—‡ ƒ† …”ƒ‹‘’Šƒ”›‰‹‘ƒǡ ‘”ƒŽ
„”‡ƒ•– ƒ† ‹˜ƒ•‹˜‡ †—…–ƒŽ …ƒ”…‹‘ƒǡ ‘”ƒŽ …‘Ž‘ ƒ† ƒ†‡‘…ƒ”…‹‘ƒǡ ‘”ƒŽ
•‡Ž‡–ƒŽ —•…Ž‡ ƒ† ”Šƒ„†‘›‘•ƒ”…‘ƒǡ ‘”ƒŽ ‹†‡› ƒ† ”‡ƒŽ …‡ŽŽ …ƒ”…‹‘ƒǡ
‘”ƒŽŽ‹˜‡”ƒ†Š‡’ƒ–‘…‡ŽŽ—Žƒ”…ƒ”…‹‘ƒǡ‘”ƒŽŽ—‰ƒ†ǡ‘”ƒŽ’ƒ…”‡ƒ•
ƒ† ƒ†‡‘…ƒ”…‹‘ƒǡ ‘”ƒŽ ’”‘•–ƒ–‡ ƒ† ƒ†‡‘…ƒ”…‹‘ƒǡ ‘”ƒŽ •‹ ƒ†
ƒŽ‹‰ƒ– ‡Žƒ‘ƒǡ ‘”ƒŽ •ƒŽŽ ‹–‡•–‹‡ ƒ† ƒŽ‹‰ƒ– ‡•‡…Š›‘ƒǡ
‘”ƒŽ •–‘ƒ…Š ƒ† ƒ†‡‘…ƒ”…‹‘ƒǡ ‘”ƒŽ —–‡”—• ƒ† ƒ†‡‘…ƒ”…‹‘ƒǡ ƒ†
‘”ƒŽ ‘˜ƒ”› ƒ† …›•–ƒ†‡‘…ƒ”…‹‘ƒǤ Š‡ –‹••—‡ •‡…–‹‘• ™‡”‡ „‘‹Ž‡† ‹ ͳͲ 
•‘†‹— …‹–”ƒ–‡ ˆ‘” ƒ–‹‰‡ ”‡–”‹‡˜ƒŽ ˆ‘ŽŽ‘™‡† „› „Ž‘…‹‰ ™‹–Š ͳͲΨ ‰‘ƒ– •‡”—ǡ
ͲǤͳΨ ”‹–‘ ǦͳͲͲ ‹ ˆ‘” ͳ Š‘—” ƒ– ”‘‘ –‡’‡”ƒ–—”‡ ȋȌǤ ‡…–‹‘• ™‡”‡
‹…—„ƒ–‡† ™‹–Š ’”‹ƒ”› ƒ–‹ǦͷŠ ’‘Ž›…Ž‘ƒŽ ƒ–‹„‘†› ȋ†‹Ž—–‹‘ ͳǣͳͲͲͲǢ …–‹˜‡
‘–‹ˆǢƒ”Ž•„ƒ†ǡȌ‹ͷΨ‰‘ƒ–•‡”—ǡͲǤͲͳΨ”‹–‘ǦͳͲͲ‹ƒ–Ͷιǡ‘˜‡”‹‰Š–Ǥ
ˆ–‡”™ƒ•Š‹‰™‹–Šƒ–ǡ•‡…–‹‘•™‡”‡‹…—„ƒ–‡†™‹–ŠŠ‘†‡†ǦǦˆˆ‹‹—”‡
…‘Œ—‰ƒ–‡† ‰‘ƒ– ƒ–‹Ǧ”ƒ„„‹– •‡…‘†ƒ”› ƒ–‹„‘†› ȋ†‹Ž—–‹‘ ͳǣʹͲͲǢ ƒ…•‘
—‘‡•‡ƒ”…ŠǢ ‡•– ”‘˜‡ǡ Ȍ ˆ‘” ͳ Š‘—” ƒ– ǡ –Š‡ ™ƒ•Š‡† ™‹–Š ƒ†
™ƒ–‡”ǡ ƒ† ‘—–‡† ™‹–Š Ž—‘”‘‘—–Ǧ
•‘Ž—–‹‘ ȋ‘—–Š‡”‹‘–‡…ŠǢ ‹”‹‰Šƒǡ
ȌǤ‹͸͹•–ƒ‹‹‰™ƒ•’‡”ˆ‘”‡†™‹–Š‹͸͹ƒ–‹„‘†›ȋŠƒ”‹‰‡Ǣƒ‹‡‰‘ǡ
Ǣ …ƒ–ƒŽ‘‰ —„‡” ͷͷͲ͸ͲͻǢ †‹Ž—–‹‘ ͳǣʹͲȌǤ Š‡ ƒ–‹Ǧͷ ƒ–‹„‘†› ™ƒ• ˆ”‘
—”‘‰‡–‡… ȋ…ƒ–ƒŽ‘‰ —„‡” ǦǦͲͳͲͲǢ †‹Ž—–‹‘ ͳǣʹͲͲȌǤ Ž‹†‡• ™‡”‡
…‘—–‡”•–ƒ‹‡†™‹–Š‘‡…Š•–͵͵ʹͷͺ†›‡ǤŽŽˆŽ‘”‡•…‡–‹ƒ‰‡•™‡”‡–ƒ‡—•‹‰ƒ
‹˜‡”–‡†Ž›’—•ͺͳˆŽ—‘”‡•…‡…‡‹…”‘•…‘’‡Ǥ
RT-PCR
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Author Manuscript Published OnlineFirst on November 3, 2011; DOI: 10.1158/0008-5472.CAN-11-2023
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
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—ƒ–‹–ƒ–‹˜‡”‡˜‡”•‡–”ƒ•…”‹’–ƒ•‡™ƒ•’‡”ˆ‘”‡†ƒ•†‡•…”‹„‡†ȋʹ͵ȌǤ
Results and Discussion
‘ †‡–‡”‹‡ –Š‡ Ž‡˜‡Ž• ‘ˆ ͷŠ† ƒ† ͷ† ‹ ‘”ƒŽ ƒ† –—‘” –‹••—‡•ǡ ™‡
†‡˜‡Ž‘’‡† ƒ •‡•‹–‹˜‡ ǦȀ ƒ••ƒ› ™‹–Š ‹•‘–‘’‡ǦŽƒ„‡Ž‡† ‹–‡”ƒŽ •–ƒ†ƒ”†•
ȋ—’’Ž‡‡–ƒ”›‹‰ǤͳȌǤͷ†™ƒ•“—ƒ–‹–ƒ–‡†™‹–Š”‡ˆ‡”‡…‡–‘–Š‡†
•–ƒ†ƒ”†Ǥ
—’’Ž‡‡–ƒ”› ‹‰Ǥ ͳ •Š‘™• ‡šƒ’Ž‡• ‘ˆ •‡’ƒ”ƒ–‹‘ ƒ† Š‘™ ƒ••
•’‡…–”‘‡–”‹… ƒƒŽ›•‹• ‘ˆ ͷŠ† ƒ† ͷ† ™ƒ• ƒ…Š‹‡˜‡†Ǥ Š‡ ‡–Š‘† ‹• •–”‹…–Ž›
“—ƒ–‹–ƒ–‹˜‡ ƒ• •Š‘™ „› •–ƒ†ƒ”† …—”˜‡• ȋ—’’Ž‡‡–ƒ”› ‹‰Ǥ ʹȌǤ ‡ ‹‹–‹ƒŽŽ›
–‡•–‡† ‹–• ’‡”ˆ‘”ƒ…‡ „› ‡ƒ•—”‹‰ ͷŠ† ƒ† ͷ† ‹ •‡˜‡”ƒŽ …‡ŽŽ ƒ† –‹••—‡
•ƒ’Ž‡• ȋ—’’Ž‡‡–ƒ”› ‹‰Ǥ ͵ȌǤ Š‡ †ƒ–ƒ ‘„–ƒ‹‡† ™‡”‡ …‘•‹•–‡– ™‹–Š
˜ƒŽ—‡•”‡’‘”–‡†‹–Š‡Ž‹–‡”ƒ–—”‡ȋʹǡͶȌƒ†™‡”‡ƒŽ•‘‰‡‡”ƒŽŽ›‹ƒ‰”‡‡‡–™‹–Šƒ
Ž‡••“—ƒ–‹–ƒ–‹˜‡‹—‘†‘–„Ž‘–ƒ••ƒ›ȋ—’’Ž‡‡–ƒ”›‹‰ǤͶȌǤ
•‹‰–Š‡ǦȀƒ••ƒ›ǡ™‡‡ƒ•—”‡†ͷŠ†‹ʹͶ•–ƒ‰‡ǦŽ—‰•
ƒ† ‹ ƒ–…Š‡† ‘”ƒŽ Ž—‰ ȋ‹‰Ǥ ͳȌǤ Š‡ Ž‡˜‡Ž• ‘ˆ ͷŠ†ǡ ‡š’”‡••‡† ƒ•
’‡”…‡– ‘ˆ †
ǡ ™‡”‡ „‡–™‡‡ ͲǤͲ͹ͺΨ ƒ† ͲǤͳͺʹΨ ‹ ‘”ƒŽ Ž—‰Ǥ  ‡˜‡”› –—‘”‡š…‡’–‘‡ȋʹȌǡ™‡•ƒ™ƒ•‹‰‹ˆ‹…ƒ–”‡†—…–‹‘‘ˆͷŠ†Ž‡˜‡Ž…‘’ƒ”‡†–‘
–Š‡ ’ƒ‹”‡† ‘”ƒŽ Ž—‰ •ƒ’Ž‡ ȋ δ ͲǤͲͷ ˆ‘” ‡ƒ…Š •ƒ’Ž‡ ’ƒ‹”Ǣ –Ǧ–‡•–Ȍ ȋ‹‰Ǥ ͳȌǤ
ͷŠ†Ž‡˜‡Ž•™‡”‡‰‡‡”ƒŽŽ›ʹǦͷǦˆ‘Ž†Ž‘™‡”‹–Š‡–—‘”•…‘’ƒ”‡†–‘‘”ƒŽŽ—‰
ȋ α ͺǤͺͺ š ͳͲǦ͹Ǣ ’ƒ‹”‡† –Ǧ–‡•–ȌǤ ‡ ƒŽ•‘ “—ƒ–‹–ƒ–‡† ͷ† ȋ‹‰Ǥ ͳȌǤ ͷ† ™ƒ•
†‡’Ž‡–‡†‹‘•––—‘”•ƒ’Ž‡•™‹–Šƒˆ‡™‡š…‡’–‹‘•ȋ–—‘”•ͳǡʹǡ͸ǡ͹ǡͳͷƒ†ͳ͸ȌǤ
ƒ›…ƒ•‡•ǡͷ†Ž‡˜‡Ž•™‡”‡Ž‘™‡”„›‘Ž›̱ͷǦʹͲΨȋαͲǤͲʹ͵Ǣ’ƒ‹”‡†–Ǧ–‡•–ȌǤ
IDH1 ‘” IDH2 —–ƒ–‹‘• ™‡”‡ ‘– ˆ‘—† ‹ –Š‡•‡ Ž—‰ –—‘”•Ǥ ‡ ƒŽ•‘ ƒƒŽ›œ‡†
ͷŠ† ‹ Ž—‰ ƒ†‡‘…ƒ”…‹‘ƒ• ƒ† ’”‹ƒ”› •ƒŽŽ …‡ŽŽ Ž—‰ …ƒ…‡”•
ȋ—’’Ž‡‡–ƒ”›‹‰ǤͷȌǤ•™‹–ŠǡͷŠ†™ƒ•†‡’Ž‡–‡†‹‘•–‘ˆ–Š‡•‡–—‘”•
”‡Žƒ–‹˜‡–‘ƒ–…Š‡†‘”ƒŽ–‹••—‡Ǥ
‡š–ǡ™‡ƒƒŽ›œ‡†–Š‡–™‘‘†‹ˆ‹‡†ʹǯǦ†‡‘š›—…Ž‡‘•‹†‡•‹•‹š‘”ƒŽ„”ƒ‹
•ƒ’Ž‡•ƒ†‹͵͵•–ƒ‰‡ƒ†ƒ•–”‘…›–‘ƒ•ȋƒ•–”‘…›–‹…‰Ž‹‘ƒ•Ȍƒ†‹–™‘
‰Ž‹‘„Žƒ•–‘ƒ•Ǥ ‡ ˆ‘—† Š‹‰Š Ž‡˜‡Ž• ‘ˆ ͷŠ† ‹ ‘”ƒŽ Š—ƒ „”ƒ‹ ’”‡ˆ”‘–ƒŽ
…‘”–‡šȋ‹‰ǤʹȌǡ‹™Š‹…ŠͷŠ†™ƒ•„‡–™‡‡ͲǤͺʹƒ†ͳǤͳͺ’‡”…‡–‘ˆ†
Ǥ‡
ƒŽ•‘‡ƒ•—”‡†Ž‡˜‡Ž•‘ˆͷŠ†ƒ†ͷ†‹ƒ•–”‘…›–‡•ƒ†‹‡—”‘•ˆ”‘Š—ƒ
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 2011 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on November 3, 2011; DOI: 10.1158/0008-5472.CAN-11-2023
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
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ˆ‡–ƒŽ „”ƒ‹Ǥ ‡˜‡Ž• ‘ˆ ͷŠ† ™‡”‡ Š‹‰Š‡” ȋͳǤͶͷΨ ͷŠ†Ȁ†
Ȍ ‹ ‡—”‘• –Šƒ ‹
ƒ•–”‘…›–‡• ȋͲǤʹ͵Ψ ͷŠ†Ȁ†
Ȍ ȋ—’’Ž‡‡–ƒ”› ‹‰Ǥ ͸ȌǤ  „”ƒ‹ –—‘”•ǡ ͷŠ†
™ƒ• •–”‘‰Ž› †‡’Ž‡–‡† ”‡Žƒ–‹˜‡ –‘ ‘”ƒŽ „”ƒ‹ ȋ‹‰Ǥ ʹȌǤ ‘‡ ƒ•–”‘…›–‘ƒ•
…‘–ƒ‹‡† ‘Ž› ͲǤͲ͵ǦͲǤͲͶΨ ‘ˆ ͷŠ†ǡ ƒ ”‡†—…–‹‘ ‘ˆ ε͵ͲǦˆ‘Ž† ȋ α ͳǤͷͷ š ͳͲǦͳͳǢ
—’ƒ‹”‡†–Ǧ–‡•–ȌǤ‹…‡ƒ•–”‘…›–‘ƒ•‹‹–‹ƒ–‡‹‡—”ƒŽ•–‡…‡ŽŽ•‘”‰Ž‹ƒŽ’”‘‰‡‹–‘”
…‡ŽŽ•ǡ–Š‡‹”†‡…”‡ƒ•‡†Ž‡˜‡Ž‘ˆͷŠ†ƒ›„‡†—‡–‘‡‹–Š‡”–Š‡ƒŽ‹‰ƒ–•–ƒ–‡‘”–‘
–Š‡ …‡ŽŽ ‘ˆ ‘”‹‰‹ ‘ˆ –Š‡•‡ –—‘”•Ǥ Š‡ ˜ƒ”›‹‰ Ž‡˜‡Ž• ‘ˆ ͷŠ† ‹ –—‘”• †‹† ‘–
…‘””‡Žƒ–‡ ™‹–Š ’ƒ–‹‡– ƒ‰‡ ‘” ™Š‡–Š‡” –Š‡ –—‘” ™ƒ• •–ƒ‰‡ ‘” •–ƒ‰‡ ‘” ™‹–Š
’ƒ–‹‡–•—”˜‹˜ƒŽȋ†ƒ–ƒ‘–•Š‘™ȌǤ‡˜‡Ž•‘ˆͷ†•Š‘™‡†‘Ž›ƒ•ƒŽŽ”‡†—…–‹‘‹
•‘‡ „”ƒ‹ –—‘”• ȋ‹‰Ǥ ʹȌ ȋ α ͲǤ͵Ǣ —’ƒ‹”‡† –Ǧ–‡•–ȌǤ Š‡”‡ ™ƒ• ‘ …‘””‡Žƒ–‹‘
„‡–™‡‡Ž‡˜‡Ž•‘ˆͷŠ†ƒ†Ž‡˜‡Ž•‘ˆͷ†Ǥ•—„•–ƒ–‹ƒŽˆ”ƒ…–‹‘‘ˆ•–ƒ‰‡ƒ†
‰Ž‹‘ƒ• …‘–ƒ‹ —–ƒ–‹‘• ‹ IDH1 ƒ† —…Š ‘”‡ ”ƒ”‡Ž›ǡ ‹ IDH2 ȋʹͶȌǤ ‡
†‡–‡”‹‡†–Š‡—–ƒ–‹‘•–ƒ–—•‘ˆͳƒ–…‘†‘ͳ͵ʹȋ—’’Ž‡‡–ƒ”›‹‰Ǥ͹ȌǤ‡
‹†‡–‹ˆ‹‡† ͳ͸ •–ƒ‰‡ Ȁ –—‘”• ™‹–Š –Š‡ –›’‹…ƒŽ …‘†‘ ͳ͵ʹ —–ƒ–‹‘ǡ ƒ† ͳ͹
•–ƒ‰‡Ȁ–—‘”•™‹–Š‘—–ƒ›ͳ—–ƒ–‹‘ǤŠ‡ͳ͵ʹͳ—–ƒ–‹‘’”‘†—…‡•
ƒ ‡‘‘”’Š‹… ‡œ›‡ ™‹–Š –Š‡ …ƒ’ƒ…‹–› –‘ ‰‡‡”ƒ–‡ ʹǦŠ›†”‘š›‰Ž—–ƒ”ƒ–‡ ȋͳͻȌǤ ‡
‡š’‡…–‡†–Šƒ–ͳ—–ƒ––—‘”•™‘—Ž†Šƒ˜‡Ž‘™‡”Ž‡˜‡Ž•‘ˆͷŠ†ƒ……‘”†‹‰–‘
–Š‡ ’”‡•—‡† ”‘Ž‡ ‘ˆ ʹǦŠ›†”‘š›‰Ž—–ƒ”ƒ–‡ ƒ• ƒ ‹Š‹„‹–‘” ‘ˆ ‘š‹†ƒ•‡• ȋsee
—’’Ž‡‡–ƒ”› ‹‰Ǥ ͺȌǤ —”’”‹•‹‰Ž›ǡ Š‘™‡˜‡”ǡ –Š‡ Ž‡˜‡Ž• ‘ˆ ͷŠ† ™‡”‡ ‡˜‡Ž›
†‹•–”‹„—–‡† „‡–™‡‡ –Š‡ Ž‘™ ƒ† Š‹‰Š ”ƒ‰‡•ǡ „‘–Š ‹ ͳ ™‹Ž†–›’‡ ƒ† ‹ ͳ
—–ƒ––—‘”•ȋ‹‰ǤʹǢαͲǤͷ͵Ǣ–Ǧ–‡•–ǡ‘Ǧ’ƒ‹”‡†ȌǤŠ‹•ˆ‹†‹‰‹•‹…‘–”ƒ•––‘ƒ
’”‡˜‹‘—• ”‡’‘”–ǡ ™Š‹…Š ‘„•‡”˜‡† ƒ •‹‰‹ˆ‹…ƒ– ”‡†—…–‹‘ ‘ˆ ͷŠ ‹ ͳǦ—–ƒ–
‰Ž‹‘ƒ•„›‹—‘Š‹•–‘…Š‡‹•–”›ȋʹͲȌǤ‹‹Žƒ”Ž›ǡͳ—–ƒ–ƒ†™‹Ž†–›’‡…ƒ•‡•
†‹†‘–•Š‘™†‹ˆˆ‡”‡…‡•‹Ž‡˜‡Ž•‘ˆͷ†ȋ‹‰ǤʹȌǤ
‘‹˜‡•–‹‰ƒ–‡‹ˆŽ‘••‘ˆͷŠ†‹•ƒˆ‡ƒ–—”‡‘ˆŠ—ƒ…ƒ…‡”•‹‰‡‡”ƒŽǡ™‡
…‘†—…–‡† ‹—‘Š‹•–‘…Š‡‹…ƒŽ •–ƒ‹‹‰ ™‹–Š ƒ ƒ–‹ǦͷŠ ƒ–‹„‘†› ȋ‹‰Ǥ ͵ ƒ†
—’’Ž‡‡–ƒ”› ‹‰Ǥ ͻȌǤ Š‹• ƒ–‹„‘†› ™ƒ• ˜‡”‹ˆ‹‡† ’”‡˜‹‘—•Ž› „› —• ƒ† —•‡† ˆ‘”
†‡–‡…–‹‰ ͷŠ ‹ ‡ƒ”Ž› ‡„”›‘• ȋͳͶǡ ʹ͵ȌǤ ‘”ƒŽ –‹••—‡ •‡…–‹‘• ƒ†
…‘””‡•’‘†‹‰ –—‘” ™‡”‡ •–ƒ‹‡† ™‹–Š –Š‹• ƒ–‹„‘†›Ǥ ‡ ‘„•‡”˜‡† •—„•–ƒ–‹ƒŽ
ͷŠ† •–ƒ‹‹‰ ‘ˆ ƒŽ‘•– ƒŽŽ …‡ŽŽ• ‹ ‘•– ‘”ƒŽ –‹••—‡•Ǥ ‘™‡˜‡”ǡ •–ƒ‹‹‰ ‹
…‘””‡•’‘†‹‰ –—‘”• ™ƒ• —‹˜‡”•ƒŽŽ› †‡…”‡ƒ•‡† ™‹–Š ‘Ž› ƒ ˆ‡™ …‡ŽŽ• ȋŽ‡•• –Šƒ
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 2011 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on November 3, 2011; DOI: 10.1158/0008-5472.CAN-11-2023
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ͺ
ͳͲΨȌ•–ƒ‹‹‰’‘•‹–‹˜‡ˆ‘”ͷŠǤŠ‡‘Ž›‡š…‡’–‹‘™ƒ•ƒ–—‘”‘”‹‰‹ƒ–‹‰‹–Š‡
…‘Ž‘ ȋ—’’Ž‡‡–ƒ”› ‹‰Ǥ ͻȌǤ ††‹–‹‘ƒŽ Ž—‰ –—‘” •Ž‹†‡• ™‡”‡ ƒŽ•‘ ƒƒŽ›œ‡†
ȋ—’’Ž‡‡–ƒ”› ‹‰Ǥ ͳͲȌ ‹…Ž—†‹‰ –—‘” ƒ† ƒ†Œƒ…‡– ‘”ƒŽ Ž—‰
ȋ—’’Ž‡‡–ƒ”›‹‰ǤͳͲȌǤ
‡ƒŽ•‘†‹†’ƒ”ƒŽŽ‡Ž•–ƒ‹‹‰‘ˆ•‡˜‡”ƒŽ‘”ƒŽƒ†–—‘”•‡…–‹‘•ˆ‘”ͷ
—•‹‰ ƒ ƒ–‹Ǧͷ ƒ–‹„‘†› ȋ—’’Ž‡‡–ƒ”› ‹‰Ǥ ͳͳȌ ƒ† †‹† ‘– ‘„•‡”˜‡ ƒ
•—„•–ƒ–‹ƒŽ †‡…”‡ƒ•‡ ‘ˆ ͷ •–ƒ‹‹‰ ‹ –Š‡ –—‘”•Ǥ Š‹• ‡ƒ• –Šƒ– –Š‡ Ž‘•• ‘ˆ
ͷŠ†‹•‘–•‹’Ž›†—‡–‘Ž‘••‘ˆͷ†‹–—‘”•Ǥ‡–Š‡†‡–‡”‹‡†‹ˆ”‡†—…‡†
•–ƒ‹‹‰‘ˆͷŠ†‹–—‘”•‹•†—‡–‘‹…”‡ƒ•‡†…‡ŽŽ’”‘Ž‹ˆ‡”ƒ–‹‘Ǥ‡—•‡†ƒƒ–‹Ǧ
‹͸͹ ƒ–‹„‘†› –‘ •–ƒ‹ ’”‘Ž‹ˆ‡”ƒ–‹‰ …‡ŽŽ•Ǥ ‡…–‹‘• ‘ˆ ‘”ƒŽ „”ƒ‹ ™‡”‡ ƒŽ‘•–
…‘’Ž‡–‡Ž›†‡˜‘‹†‘ˆ‹͸͹ƒ–‹‰‡„—–„”ƒ‹–—‘”•…‘–ƒ‹‡†ƒ›‹͸͹Ǧ’‘•‹–‹˜‡
…‡ŽŽ•–Šƒ–Žƒ…‡†ͷŠ•–ƒ‹‹‰ȋ—’’Ž‡‡–ƒ”›‹‰ǤͳʹȌǤ‹‹Žƒ”Ž›ǡ–Š‡”‡™ƒ•Ž‹––Ž‡
‹͸͹ •–ƒ‹‹‰ ‹ ‘”ƒŽ Ž—‰ǡ „—– ‹ ƒ†Œƒ…‡– …ƒ”…‹‘ƒ –‹••—‡ǡ ™‡ •ƒ™ —–—ƒŽŽ›
‡š…Ž—•‹˜‡•–ƒ‹‹‰‘ˆ‹͸͹ƒ†ͷŠȋ‹‰ǤͶǢ—’’Ž‡‡–ƒ”›‹‰ǤͳͲȌǤŠ‡•ƒ‡™ƒ•
ˆ‘—†ˆ‘”•‡…–‹‘•‘ˆ„”‡ƒ•–ǡ’ƒ…”‡ƒ–‹…ƒ†—–‡”—•–—‘”•ȋ—’’Ž‡‡–ƒ”›‹‰Ǥͳ͵ȌǤ
‹••—‡ •‡…–‹‘• ‘ˆ ‘”ƒŽ •ƒŽŽ ‹–‡•–‹‡ •Š‘™‡† •–”‘‰ ‹͸͹ •–ƒ‹‹‰ ˆ‘”
’”‘Ž‹ˆ‡”ƒ–‹‰ …‡ŽŽ• ƒ––Š‡ „‘––‘ ‘ˆ …”›’–• ™‹–Š Žƒ… ‘ˆ ͷŠ •–ƒ‹‹‰ǡ ™Š‡”‡ƒ• –Š‡
‘”‡ †‹ˆˆ‡”‡–‹ƒ–‡† …‡ŽŽ• …‘–ƒ‹‡† Š‹‰Š Ž‡˜‡Ž• ‘ˆ ͷŠ ƒ† Žƒ…‡† ‹͸͹ •–ƒ‹‹‰
ȋ‹‰Ǥ ͶȌǤ Š—•ǡ ‘‡ •–”ƒ‹‰Š–ˆ‘”™ƒ”† ‡š’Žƒƒ–‹‘ ˆ‘” –Š‡ Ž‘•• ‘ˆ ͷŠ† ‹ –—‘”• ‹•
–Š‡‡Šƒ…‡†”ƒ–‡‘ˆ…‡ŽŽ’”‘Ž‹ˆ‡”ƒ–‹‘‹–—‘”•–Šƒ–…‘—Ž†Ž‡ƒ†–‘ƒ’ƒ••‹˜‡Ž‘••‘ˆ
ͷŠ†ǡ™Š‹…Š‹•‘–ƒ•—„•–”ƒ–‡ˆ‘”ͳȋʹͷȌǤ‡‘–‹…‡†–Šƒ–‘–ƒŽŽ…‡ŽŽ•–Šƒ–
Žƒ… ͷŠ •–ƒ‹‹‰ ‹ –Š‡ –—‘”• ƒ”‡ ‹͸͹Ǧ’‘•‹–‹˜‡ ’‡”Šƒ’• †—‡ –‘ ’ƒ•– Š‹•–‘”› ‘ˆ
’”‘Ž‹ˆ‡”ƒ–‹‘Ž‡ƒ†‹‰–‘’‡”ƒ‡–Ž‘••‘ˆͷŠǤ
Š‡‹˜‡”•‡Ž‹„‡–™‡‡ͷŠŽ‡˜‡Ž•ƒ†…‡ŽŽ’”‘Ž‹ˆ‡”ƒ–‹‘ƒ›ƒŽ•‘”‡ˆŽ‡…–
‹–”‹•‹… †‹ˆˆ‡”‡…‡• ‹ …‡ŽŽ –›’‡ǡ ƒ• ™‡ ƒ”‡ …‘’ƒ”‹‰ †‹ˆˆ‡”‡–‹ƒ–‡† …‡ŽŽ• ™‹–Š
—†‹ˆˆ‡”‡–‹ƒ–‡† …ƒ…‡” …‡ŽŽ• –Šƒ– …ƒ””› •–‡ …‡ŽŽǦŽ‹‡ ’”‘’‡”–‹‡• ‘” ‘”‹‰‹ƒ–‡ ˆ”‘
•‘ƒ–‹…•–‡…‡ŽŽ•Ǥ‡‘–Š‡”ƒŽ–‡”ƒ–‹˜‡‹•–Š‡’‘••‹„Ž‡‡š‹•–‡…‡‘ˆƒ„‡””ƒ–‹‘•‹
ͷŠ’”‘†—…–‹‘‘”‡Ž‹‹ƒ–‹‘’ƒ–Š™ƒ›•‹–—‘”•Ǥ—–ƒ–‹‘•‹TET‰‡‡•Šƒ˜‡
‘–„‡‡”‡’‘”–‡†‹•‘Ž‹†–—‘”•ǤŠ‡”‡™ƒ•‘•—„•–ƒ–‹ƒŽ”‡†—…–‹‘‘ˆTET‰‡‡
‡š’”‡••‹‘‹Ž—‰ƒ†„”ƒ‹–—‘”•”‡Žƒ–‹˜‡–‘‘”ƒŽ–‹••—‡ƒ•…‘ˆ‹”‡†„›Ǧ
ȋ—’’Ž‡‡–ƒ”›‹‰ǤͳͶȌǤ
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 2011 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on November 3, 2011; DOI: 10.1158/0008-5472.CAN-11-2023
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ͻ
Š‡Ž‘••‘ˆͷŠ†‹–—‘”•ƒ›Šƒ˜‡’”‘ˆ‘—†‡ˆˆ‡…–•‘‡–Š›Žƒ–‹‘
’ƒ––‡”•Ǥ‘”‡šƒ’Ž‡ǡ‹ˆͷŠ‹•ƒ‹–‡”‡†‹ƒ–‡‹†‡‡–Š›Žƒ–‹‘ǡ‹–•Ž‘••ƒ–
•’‡…‹ˆ‹… ‰‡‘‹… Ž‘…ƒ–‹‘• ƒ› ƒ‡ –Š‡•‡ •‡“—‡…‡• ‘”‡ ’”‘‡ –‘ ƒ…“—‹”‡
‡–Š›Žƒ–‹‘Ǥ–™‹ŽŽ„‡‡••‡–‹ƒŽ–‘—†‡”•–ƒ†–Š‡‡…Šƒ‹••‘ˆŠ‘™ͷŠ‹•Ž‘•–
‹–—‘”•Ǥ‹ƒŽŽ›ǡŽ‘••‘ˆͷǦŠ›†”‘š›‡–Š›Ž…›–‘•‹‡…‘—Ž†„‡…‘‡ƒ—•‡ˆ—Ž‘Ž‡…—Žƒ”
„‹‘ƒ”‡” ˆ‘” …ƒ…‡” †‡–‡…–‹‘ ƒ† †‹ƒ‰‘•‹•ǡ ’‡”Šƒ’• ‹ …‘Œ—…–‹‘ ™‹–Š ‹͸͹
•–ƒ‹‹‰Ǥ
Acknowledgementsǣ
Š‹• ™‘” ™ƒ• •—’’‘”–‡† „› ‰”ƒ–• ͲͺͶͶ͸ͻ ƒ† Ͳ͵͸ͲͶͳ –‘ ǤǤǡ
Ͳ͹ͷ͵ͻ͵–‘ǤǤǡƒ†ͳͲͳͺ͸Ͷ–‘ǤǤ
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 2011 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on November 3, 2011; DOI: 10.1158/0008-5472.CAN-11-2023
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ͳͲ
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”‹ƒ—…‹‘‹• ǡ ‡‹–œ Ǥ Š‡ —…Ž‡ƒ” „ƒ•‡ ͷǦŠ›†”‘š›‡–Š›Ž…›–‘•‹‡ ‹•
’”‡•‡–‹—”‹Œ‡‡—”‘•ƒ†–Š‡„”ƒ‹Ǥ…‹‡…‡ʹͲͲͻǢ͵ʹͶǣͻʹͻǦͻ͵ͲǤ
ƒŠ‹Ž‹ƒ‹ ǡ ‘Š ǡ Š‡ ǡ ƒ•–‘” ǡ ƒ†—™ƒŽƒ ǡ ”—†‘ , et al.
‘˜‡”•‹‘ ‘ˆ ͷǦ‡–Š›Ž…›–‘•‹‡ –‘ ͷǦŠ›†”‘š›‡–Š›Ž…›–‘•‹‡ ‹ ƒƒŽ‹ƒ
„›’ƒ”–‡”ͳǤ…‹‡…‡ʹͲͲͻǢ͵ʹͶǣͻ͵ͲǦͻ͵ͷǤ
–‘ ǡ ̵Ž‡••‹‘ ǡ ƒ”ƒ‘˜ƒ ǡ ‘‰ ǡ ‘™‡”• ǡ Šƒ‰ Ǥ ‘Ž‡ ‘ˆ ‡–
’”‘–‡‹•‹ͷ–‘ͷŠ…‘˜‡”•‹‘ǡǦ…‡ŽŽ•‡ŽˆǦ”‡‡™ƒŽƒ†‹‡”…‡ŽŽƒ••
•’‡…‹ˆ‹…ƒ–‹‘Ǥƒ–—”‡ʹͲͳͲǢ‹’”‡••Ǥ
Ž‘„‹•…Šǡ—œ‡Žǡ—ŽŽ‡”ǡ‹…ŠƒŽƒ‹•ǡƒ‰‡”ǡ‘…Š, et al.‹••—‡
†‹•–”‹„—–‹‘‘ˆͷǦŠ›†”‘š›‡–Š›Ž…›–‘•‹‡ƒ†•‡ƒ”…Šˆ‘”ƒ…–‹˜‡†‡‡–Š›Žƒ–‹‘
‹–‡”‡†‹ƒ–‡•Ǥ‘‡ʹͲͳͲǢͷǣ‡ͳͷ͵͸͹Ǥ
œ™ƒ‰‹‡”…œƒ ǡ —Ž–ƒ ǡ …Š‹†– ǡ ’ƒ†ƒ ǡ ‡‘Šƒ”†– Ǥ ‡•‹–‹˜‡
‡œ›ƒ–‹… “—ƒ–‹ˆ‹…ƒ–‹‘ ‘ˆ ͷǦŠ›†”‘š›‡–Š›Ž…›–‘•‹‡ ‹ ‰‡‘‹… Ǥ
—…Ž‡‹……‹†•‡•ʹͲͳͲǢ͵ͺǣ‡ͳͺͳǤ
— ǡ Šƒ‰ Ǥ …–‹˜‡ †‡‡–Š›Žƒ–‹‘ǣ ƒ› ”‘ƒ†• Ž‡ƒ† –‘ ‘‡Ǥ ƒ–
‡˜‘Ž‡ŽŽ‹‘ŽʹͲͳͲǢͳͳǣ͸Ͳ͹Ǧ͸ʹͲǤ
‹…œ ǡ ”ƒ…‘ ǡ ‡‹•‡„‡”‰‡” ǡ ƒ–‘• ǡ ”—‡‰‡” ǡ ‘”‡ , et al.
›ƒ‹… ”‡‰—Žƒ–‹‘ ‘ˆ ͷǦŠ›†”‘š›‡–Š›Ž…›–‘•‹‡ ‹ ‘—•‡ …‡ŽŽ• ƒ†
†—”‹‰†‹ˆˆ‡”‡–‹ƒ–‹‘Ǥƒ–—”‡ʹͲͳͳǢͶ͹͵ǣ͵ͻͺǦͶͲʹǤ
‘Šǡƒ„——…Š‹ǡƒ‘ǡ—ƒ‰ǡ—‹ˆˆǡƒ”†‘‡, et al.‡–ͳƒ†‡–ʹ
”‡‰—Žƒ–‡ ͷǦŠ›†”‘š›‡–Š›Ž…›–‘•‹‡ ’”‘†—…–‹‘ ƒ† …‡ŽŽ Ž‹‡ƒ‰‡ •’‡…‹ˆ‹…ƒ–‹‘
‹‘—•‡‡„”›‘‹…•–‡…‡ŽŽ•Ǥ‡ŽŽ–‡‡ŽŽʹͲͳͳǢͺǣʹͲͲǦʹͳ͵Ǥ
ƒ•–‘”ǡƒ’‡ǡ—ƒ‰ǡ‡†‡”•‘ǡ‹•–‡”ǡ‘, et al.
‡‘‡Ǧ
™‹†‡ ƒ’’‹‰ ‘ˆ ͷǦŠ›†”‘š›‡–Š›Ž…›–‘•‹‡ ‹ ‡„”›‘‹… •–‡ …‡ŽŽ•Ǥ ƒ–—”‡
ʹͲͳͳǢͶ͹͵ǣ͵ͻͶǦ͵ͻ͹Ǥ
—ǡ̵Ž‡••‹‘ǡ–‘ǡƒ‰ǡ—‹ǡŠƒ‘, et al.
‡‘‡Ǧ™‹†‡ƒƒŽ›•‹•
‘ˆ ͷǦŠ›†”‘š›‡–Š›Ž…›–‘•‹‡ †‹•–”‹„—–‹‘ ”‡˜‡ƒŽ• ‹–• †—ƒŽ ˆ—…–‹‘ ‹
–”ƒ•…”‹’–‹‘ƒŽ ”‡‰—Žƒ–‹‘ ‹ ‘—•‡ ‡„”›‘‹… •–‡ …‡ŽŽ•Ǥ ‡‡• ‡˜
ʹͲͳͳǢʹͷǣ͸͹ͻǦ͸ͺͶǤ
‹ŽŽ‹ƒ•ǡŠ”‹•–‡•‡ǡ‡†‡”•‡ǡ‘Šƒ•‡ǡŽ‘‘•ǡƒ’’•‹Ž„‡”,
et al.ͳƒ†Š›†”‘š›‡–Š›Ž…›–‘•‹‡‹–”ƒ•…”‹’–‹‘ƒ†‡–Š›Žƒ–‹‘
ˆ‹†‡Ž‹–›Ǥƒ–—”‡ʹͲͳͳǢͶ͹͵ǣ͵Ͷ͵Ǧ͵ͶͺǤ
—ǡ—ǡƒǡ‘‰ǡ‹‘‰ǡ‡‰, et al.
‡‘‡Ǧ™‹†‡”‡‰—Žƒ–‹‘‘ˆ
ͷŠǡ ͷǡ ƒ† ‰‡‡ ‡š’”‡••‹‘ „› ‡–ͳ Š›†”‘š›Žƒ•‡ ‹ ‘—•‡ ‡„”›‘‹…
•–‡…‡ŽŽ•Ǥ‘Ž‡ŽŽʹͲͳͳǢͶʹǣͶͷͳǦͶ͸ͶǤ
‘‰ǡœ—Ž™ƒ…Šǡ—ǡƒ‹ǡ‹ǡ‹, et al.‡Ž‡…–‹˜‡…Š‡‹…ƒŽŽƒ„‡Ž‹‰
”‡˜‡ƒŽ• –Š‡ ‰‡‘‡Ǧ™‹†‡ †‹•–”‹„—–‹‘ ‘ˆ ͷǦŠ›†”‘š›‡–Š›Ž…›–‘•‹‡Ǥ ƒ–
‹‘–‡…Š‘ŽʹͲͳͳǢʹͻǣ͸ͺǦ͹ʹǤ
‹
ǡ—ǡ‹ǡˆ‡‹ˆ‡”
Ǥ
‡‘‹…ƒ’’‹‰‘ˆͷǦŠ›†”‘š›‡–Š›Ž…›–‘•‹‡
‹–Š‡Š—ƒ„”ƒ‹Ǥ—…Ž‡‹……‹†•‡•ʹͲͳͳǢ͵ͻǣͷͲͳͷǦͷͲʹͶǤ
‡ŽŠ‘‡ƒ— ǡ —’‘– ǡ ‡ŽŽƒ ƒŽŽ‡ ǡ ƒ‡• ǡ ”ƒ‘› ǡ ƒ••‡ , et al.
—–ƒ–‹‘‹ʹ‹›‡Ž‘‹†…ƒ…‡”•Ǥ‰Ž‡†ʹͲͲͻǢ͵͸ͲǣʹʹͺͻǦʹ͵ͲͳǤ
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 2011 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on November 3, 2011; DOI: 10.1158/0008-5472.CAN-11-2023
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ͳ͸Ǥ
ͳ͹Ǥ
ͳͺǤ
ͳͻǤ
ʹͲǤ
ʹͳǤ
ʹʹǤ
ʹ͵Ǥ
ʹͶǤ
ʹͷǤ
ͳͳ
‘ ǡ —ƒ‰ ǡ ƒ‘™•ƒ ǡ ƒ’‡ ǡ ƒŠ‹Ž‹ƒ‹ ǡ ƒ†—™ƒŽƒ , et al.
’ƒ‹”‡†Š›†”‘š›Žƒ–‹‘‘ˆͷǦ‡–Š›Ž…›–‘•‹‡‹›‡Ž‘‹†…ƒ…‡”•™‹–Š—–ƒ–
ʹǤƒ–—”‡ʹͲͳͲǢͶ͸ͺǣͺ͵ͻǦͺͶ͵Ǥ
‹‰—‡”‘ƒǡ„†‡ŽǦƒŠƒ„ǡ—ǡƒ”†ǡƒ–‡ŽǡŠ‹Š, et al.‡—‡‹…
ͳ ƒ† ʹ —–ƒ–‹‘• ”‡•—Ž– ‹ ƒ Š›’‡”‡–Š›Žƒ–‹‘ ’Š‡‘–›’‡ǡ †‹•”—’–
ʹ ˆ—…–‹‘ǡ ƒ† ‹’ƒ‹” Š‡ƒ–‘’‘‹‡–‹… †‹ˆˆ‡”‡–‹ƒ–‹‘Ǥ ƒ…‡” ‡ŽŽ
ʹͲͳͲǢͳͺǣͷͷ͵Ǧͷ͸͹Ǥ
‘—•Š‡Š”ǡ‡‹•‡„‡”‰‡”ǡ‹‡ˆ‡•ǡŠ‹ŽŽ‹’•ǡ—Œƒ”ƒǡ‡”ƒ,
et al. †‡–‹ˆ‹…ƒ–‹‘ ‘ˆ ƒ ’
‹•Žƒ† ‡–Š›Žƒ–‘” ’Š‡‘–›’‡ –Šƒ– †‡ˆ‹‡• ƒ
†‹•–‹…–•—„‰”‘—’‘ˆ‰Ž‹‘ƒǤƒ…‡”‡ŽŽʹͲͳͲǢͳ͹ǣͷͳͲǦͷʹʹǤ
ƒ‰ ǡ Š‹–‡ ǡ ”‘•• ǡ ‡‡–– ǡ ‹––‹‰‡” ǡ ”‹‰‰‡”• , et al.
ƒ…‡”Ǧƒ••‘…‹ƒ–‡† ͳ —–ƒ–‹‘• ’”‘†—…‡ ʹǦŠ›†”‘š›‰Ž—–ƒ”ƒ–‡Ǥ ƒ–—”‡
ʹͲͲͻǢͶ͸ʹǣ͹͵ͻǦ͹ͶͶǤ
— ǡ ƒ‰ ǡ ‹— ǡ ƒ‰ ǡ ƒ‰ ǡ ‹ , et al. …‘‡–ƒ„‘Ž‹–‡ ʹǦ
Š›†”‘š›‰Ž—–ƒ”ƒ–‡‹•ƒ…‘’‡–‹–‹˜‡‹Š‹„‹–‘”‘ˆƒŽ’ŠƒǦ‡–‘‰Ž—–ƒ”ƒ–‡Ǧ†‡’‡†‡–
†‹‘š›‰‡ƒ•‡•Ǥƒ…‡”‡ŽŽʹͲͳͳǢͳͻǣͳ͹Ǧ͵ͲǤ
ƒ‘ ǡ ƒ‰ Ǥ ‘ŽŽ‹•‹‘ƒŽŽ› ƒ…–‹˜ƒ–‡† †‹••‘…‹ƒ–‹‘ ‘ˆ ’”‘–‘ƒ–‡† ʹ̵Ǧ
†‡‘š›…›–‹†‹‡ǡʹ̵Ǧ†‡‘š›—”‹†‹‡ǡƒ†–Š‡‹”‘š‹†ƒ–‹˜‡Ž›†ƒƒ‰‡††‡”‹˜ƒ–‹˜‡•Ǥ
‘…ƒ••’‡…–”‘ʹͲͲ͸Ǣͳ͹ǣͳ͵͵ͷǦͳ͵ͶͳǤ
ƒ”ƒ…‘‹• ǡ —Œ‹‘–‘ ǡ ‘™‡”• Ǥ ›–Š‡•‹• ƒ† …Šƒ”ƒ…–‡”‹œƒ–‹‘ ‘ˆ
‹•‘–‘’‹…ƒŽŽ› ‡”‹…Š‡† ’›”‹‹†‹‡ †‡‘š›—…Ž‡‘•‹†‡ ‘š‹†ƒ–‹‘ †ƒƒ‰‡
’”‘†—…–•ǤŠ‡‡•‘š‹…‘ŽͳͻͻͺǢͳͳǣ͹ͷǦͺ͵Ǥ
“„ƒŽǡ‹
ǡˆ‡‹ˆ‡”
ǡœƒ„‘Ǥ‡’”‘‰”ƒ‹‰‘ˆ–Š‡’ƒ–‡”ƒŽ‰‡‘‡
—’‘ˆ‡”–‹Ž‹œƒ–‹‘‹˜‘Ž˜‡•‰‡‘‡Ǧ™‹†‡‘š‹†ƒ–‹‘‘ˆͷǦ‡–Š›Ž…›–‘•‹‡Ǥ”‘…
ƒ–Ž…ƒ†…‹ʹͲͳͳǢͳͲͺǣ͵͸ͶʹǦ͵͸Ͷ͹Ǥ
ƒǡƒ”•‘•ǡ‹
ǡ…‡†‘ǡƒ•Š‡‡†ǡ—ƒ, et al.ͳƒ†
ʹ—–ƒ–‹‘•‹‰Ž‹‘ƒ•Ǥ‰Ž‡†ʹͲͲͻǢ͵͸Ͳǣ͹͸ͷǦ͹͹͵Ǥ
ƒŽ‹Ž—…ǡ‘™‡”•Ǥ†‘‰‡‘—•…›–‘•‹‡†ƒƒ‰‡’”‘†—…–•ƒŽ–‡”–Š‡•‹–‡
•‡Ž‡…–‹˜‹–› ‘ˆ Š—ƒ ƒ‹–‡ƒ…‡ ‡–Š›Ž–”ƒ•ˆ‡”ƒ•‡ ͳǤ ƒ…‡”
‡•ʹͲͲ͹Ǣ͸͹ǣͻͶ͸ǦͻͷͲǤ
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 2011 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on November 3, 2011; DOI: 10.1158/0008-5472.CAN-11-2023
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ͳʹ
Figure Legends
Figure 1:
Quantitation of 5hmdC and 5mdC in normal lung and lung squamous cell
carcinoma DNA. A. ͷŠ†Ǥ Š‡ ˆ‹”•– ͳͺ •ƒ’Ž‡• ƒ”‡ ƒ–…Š‡† ‘”ƒŽ Ž—‰ ȋǡ
„Ž—‡Ȍ ƒ† Ž—‰ –—‘”• ȋǡ ‰”‡‡ȌǤ Š‡ Žƒ•– •‹š •ƒ’Ž‡• ƒ”‡ Ž—‰ –—‘”• ™‹–Š‘—–
ƒ˜ƒ‹Žƒ„Ž‡ ‘”ƒŽ –‹••—‡Ǥ B. ͷ†Ǥ Š‡ ƒ•–‡”‹•• ‹†‹…ƒ–‡ –Šƒ– –Š‡ Ž‡˜‡Ž• ‘ˆ ͷ†
™‡”‡•‹‰‹ˆ‹…ƒ–Ž›”‡†—…‡†‹–Š‡–—‘”…‘’ƒ”‡†–‘‘”ƒŽŽ—‰ȋδͲǤͲͷȌǤ
Figure 2:
Quantitation of 5hmdC and 5mdC in normal brain DNA and in stage II/III
astrocytomas. A. ͷŠ† “—ƒ–‹–ƒ–‹‘ ‹ ‘”ƒŽ „”ƒ‹ ȋǡ „Ž—‡Ȍ ƒ† ‹ „”ƒ‹
–—‘”•ȋǡ‰”‡‡‘”‘”ƒ‰‡ȌǤƒ’Ž‡•ͳǦͳ͸ǡʹͷǡʹ͹Ǧʹͻƒ†͵ʹǦ͵͸™‡”‡
•–ƒ‰‡ƒ•–”‘…›–‘ƒ•Ǣͳ͹ǦʹͶǡʹ͸ǡ͵Ͳƒ†͵ͳ™‡”‡•–ƒ‰‡ƒ•–”‘…›–‘ƒ•ǡ
ƒ†͵͹ƒ†͵ͺ™‡”‡‰Ž‹‘„Žƒ•–‘ƒ•ǤB.ͷ†‹‘”ƒŽ„”ƒ‹ȋȌƒ†‹„”ƒ‹
–—‘”•ȋȌǤ—‘”•™‹–Š‘ͳ—–ƒ–‹‘ƒ”‡•Š‘™‹‰”‡‡Ǣ–—‘”•™‹–Šͳ
ͳ͵ʹƒ”‡•Š‘™‹‘”ƒ‰‡ǤŠ‡•ƒ’Ž‡ʹ͸Šƒ†ƒ‹‘”ƒŽŽ‡Ž‡ˆ”‡“—‡…›‘ˆͳ
ͳ͵ʹǤƒ’Ž‡ʹͷŠƒ†–Š‡”ƒ”‡—–ƒ–‹‘ͳ͵ʹ
Ǥ
Figure 3:
Immunohistochemical analysis of 5hmC on human tissue arrays. —ƒ–‹••—‡
ƒ””ƒ›• …‘–ƒ‹‹‰ •ƒ’Ž‡• ‘ˆ ƒŽ‹‰ƒ– –—‘” ƒ† …‘””‡•’‘†‹‰ ‘”ƒŽ –‹••—‡
™‡”‡•–ƒ‹‡†™‹–Šƒ–‹ǦͷŠƒ–‹„‘†›Ǥ–ƒ‹‹‰™‹–Š‘‡…Š•–͵͵ʹͷͺ‹••Š‘™ƒ•ƒ
…‘–”‘ŽǤŠ‡ƒ‰‹ˆ‹…ƒ–‹‘‘ˆƒŽŽ’ƒ‡Ž•‹•ͳͲǦˆ‘Ž†Ǥ
Figure 4:
Inverse relationship between 5hmC and Ki67 staining. ‡…–‹‘•‘ˆ‘”ƒŽŽ—‰
ƒ† Ž—‰ –—‘”ǡ ‘”ƒŽ ’”‘•–ƒ–‡ ƒ† ’”‘•–ƒ–‡ –—‘” ƒ† ‘”ƒŽ •ƒŽŽ ‹–‡•–‹‡
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ͳ͵
™‡”‡ •–ƒ‹‡† ™‹–Š ƒ–‹ǦͷŠ ȋ”‡†Ȍ ƒ† ƒ–‹Ǧ‹͸͹ ƒ–‹„‘†‹‡• ȋ‰”‡‡Ȍ –‘ ƒ”
’”‘Ž‹ˆ‡”ƒ–‹‰ …‡ŽŽ•Ǥ ‘–‡ –Š‡ —–—ƒŽŽ› ‡š…Ž—•‹˜‡ •–ƒ‹‹‰ ‘ˆ ͷŠ ƒ† ‹͸͹ ‹ –Š‡
–‹••—‡ •‡…–‹‘•Ǥ ‘” ‡šƒ’Ž‡ǡ ’”‘Ž‹ˆ‡”ƒ–‹‰ …‡ŽŽ• ‹ ‹–‡•–‹ƒŽ …”›’–•ƒ”‡ ’‘•‹–‹˜‡ ˆ‘”
‹͸͹„—–‡‰ƒ–‹˜‡ˆ‘”ͷŠǤ
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5-hydroxymethylcytosine is strongly depleted in human cancers
but its levels do not correlate with IDH1 mutations
Seung-gi Jin, Yong Jiang, Runxiang Qiu, et al.
Cancer Res Published OnlineFirst November 3, 2011.
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