Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Inhibition of AMPK and Krebs cycle gene expression drives metabolic
remodeling of Pten-deficient preneoplastic thyroid cells
ƒŽ‡”‹ƒ
Ǥ–‹…‘”…‹—…Šǡƒ”‹ƒǤ—••‘ǡ”‹•–›Ǥƒ‰ǡƒ†–‘‹‘‹”‹•–‘ˆƒ‘
‡’ƒ”–‡– ‘ˆ ‡˜‡Ž‘’‡–ƒŽ ƒ† ‘Ž‡…—Žƒ” ‹‘Ž‘‰›ǡ Ž„‡”– ‹•–‡‹ ‘ŽŽ‡‰‡ ‘ˆ
‡†‹…‹‡ǡ”‘šǡǡǤ
—‹‰‹–Ž‡ǣ‘˜‡Ž‡–ƒ„‘Ž‹•Ǧ”‡‘†‡Ž‹‰’ƒ–Š™ƒ›
‡›™‘”†•ǣŠ›”‘‹†ǡ–‡ǡǡ…›…Ž‡ǡ‘—•‡‘†‡Ž
‘””‡•’‘†‹‰ ƒ—–Š‘”ǣ Ǥ ‹ ”‹•–‘ˆƒ‘ǡ ‡’ƒ”–‡– ‘ˆ ‡˜‡Ž‘’‡–ƒŽ ƒ† ‘Ž‡…—Žƒ”
‹‘Ž‘‰›ǡŽ„‡”–‹•–‡‹‘ŽŽ‡‰‡‘ˆ‡†‹…‹‡ǡ”‹…‡‡–‡”ˆ‘”
‡‡–‹…ƒ†”ƒ•Žƒ–‹‘ƒŽ
‡†‹…‹‡ǡͳ͵Ͳͳ‘””‹•ƒ”˜‡—‡ǡ‘‘͵Ͳʹǡ”‘šǡͳͲͶ͸ͳǤ
‡Žǣ͹ͳͺǦ͸͹ͺǦͳͳ͵͹Ǣƒšǣ͹ͳͺǦ͸͹ͺǦͳͲʹͲ
ƒ‹Žǣƒ–‘‹‘Ǥ†‹…”‹•–‘ˆƒ‘̷‡‹•–‡‹Ǥ›—Ǥ‡†—
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ABSTRACT
ƒ’‹†Ž›’”‘Ž‹ˆ‡”ƒ–‹‰ƒ†‡‘’Žƒ•–‹…ƒŽŽ›–”ƒ•ˆ‘”‡†…‡ŽŽ•‰‡‡”ƒ–‡–Š‡‡‡”‰›”‡“—‹”‡†
–‘ •—’’‘”– ”ƒ’‹† …‡ŽŽ †‹˜‹•‹‘ „› ‹…”‡ƒ•‹‰ ‰Ž›…‘Ž›•‹• ƒ† †‡…”‡ƒ•‹‰ ˆŽ—š –Š”‘—‰Š –Š‡
‘š‹†ƒ–‹˜‡ ’Š‘•’Š‘”›Žƒ–‹‘ ’ƒ–Š™ƒ› ȋȌǡ —•—ƒŽŽ› ™‹–Š‘—– ƒŽ–‡”ƒ–‹‘• ‹
‹–‘…Š‘†”‹ƒŽ ˆ—…–‹‘Ǥ  …‘–”ƒ•–ǡ Ž‹––Ž‡ ‹• ‘™ ‘ˆ –Š‡ ‡–ƒ„‘Ž‹… ƒŽ–‡”ƒ–‹‘•ǡ ‹ˆ ƒ›ǡ
™Š‹…Š‘……—”‹…‡ŽŽ•Šƒ”„‘”‹‰—–ƒ–‹‘•–Šƒ–’”‹‡–Š‡‹”‡‘’Žƒ•–‹…–”ƒ•ˆ‘”ƒ–‹‘Ǥ‘
ƒ††”‡•• –Š‹• “—‡•–‹‘ǡ ™‡ —•‡† ƒ PtenǦ†‡ˆ‹…‹‡– ‘—•‡ ‘†‡Ž –‘ ‡šƒ‹‡ –Š›”‘‹† …‡ŽŽ•
™Š‡”‡ ƒ ‹Ž† Š›’‡”’Žƒ•‹ƒ ’”‘‰”‡••‡• •Ž‘™Ž› –‘ ˆ‘ŽŽ‹…—Žƒ” –Š›”‘‹† …ƒ”…‹‘ƒǤ •‹‰ –Š‹•
‘†‡Žǡ ™‡ ”‡’‘”– –Šƒ– …‘•–‹–—–‹˜‡ ͵ ƒ…–‹˜ƒ–‹‘ …ƒ—•‡† „› †‡ˆ‹…‹‡…› ‹ ‘Ǧ
–”ƒ•ˆ‘”‡†–Š›”‘…›–‡•”‡•—Ž–•‹ƒ‰Ž‘„ƒŽ†‘™Ǧ”‡‰—Žƒ–‹‘‘ˆ”‡„•…›…Ž‡ƒ†
‰‡‡ ‡š’”‡••‹‘ǡ †‡ˆ‡…–‹˜‡ ‹–‘…Š‘†”‹ƒǡ ”‡†—…‡† ”‡•’‹”ƒ–‹‘ ƒ† ƒ ‡Šƒ…‡‡– ‹
…‘’‡•ƒ–‘”› ‰Ž›…‘Ž›•‹•Ǥ ‡ ˆ‘—† –Šƒ– –Š‹• ’”‘…‡•• †‘‡• ‘– ‹˜‘Ž˜‡ ƒ› ‘ˆ –Š‡
’ƒ–Š™ƒ›• …Žƒ••‹…ƒŽŽ› ƒ••‘…‹ƒ–‡† ™‹–Š –Š‡ ƒ”„—”‰ ‡ˆˆ‡…–Ǥ ‘”‡‘˜‡”ǡ –Š‹• ’”‘…‡•• ™ƒ•
‹†‡’‡†‡– ‘ˆ ’”‘Ž‹ˆ‡”ƒ–‹‘ „—– …‘–”‹„—–‡† †‹”‡…–Ž› –‘ –Š›”‘‹† Š›’‡”’Žƒ•‹ƒǤ —”
ˆ‹†‹‰•†‡ˆ‹‡ƒ‘˜‡Ž‡–ƒ„‘Ž‹…•™‹–…Š–‘‰Ž›…‘Ž›•‹•†”‹˜‡„›͵Ǧ†‡’‡†‡–
‹ƒ…–‹˜ƒ–‹‘ ™‹–Š ƒ …‘•‡“—‡– ”‡’”‡••‹‘ ‹ –Š‡ ‡š’”‡••‹‘ ‘ˆ ‡› ‡–ƒ„‘Ž‹…
–”ƒ•…”‹’–‹‘”‡‰—Žƒ–‘”•Ǥ
ʹ
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
INTRODUCTION
‹‰ŠŽ› ’”‘Ž‹ˆ‡”ƒ–‹˜‡ …‘†‹–‹‘•ǡ •—…Š ƒ• ‡„”›‘‹… †‡˜‡Ž‘’‡–ǡ –‹••—‡ ”‡‰‡‡”ƒ–‹‘ǡ
Ž›’Š‘…›–‡ƒ…–‹˜ƒ–‹‘ǡƒ†‡‘’Žƒ•–‹…–”ƒ•ˆ‘”ƒ–‹‘ǡ”‡“—‹”‡ƒ…‘’Ž‡š”‡‘”‰ƒ‹œƒ–‹‘
‘ˆ ‡‡”‰› ‡–ƒ„‘Ž‹• –‘ ˆ‡‡† …‡ŽŽ ‰”‘™–Š ƒ† †‹˜‹•‹‘ ȋͳȌǤ ƒŒ‘” ˆ‡ƒ–—”‡ ‘ˆ –Š‹•
‡–ƒ„‘Ž‹…”‡‘†‡Ž‹‰‹•–Š‡•™‹–…Šˆ”‘‘š‹†ƒ–‹˜‡’Š‘•’Š‘”›Žƒ–‹‘–‘ƒ‡”‘„‹…‰Ž›…‘Ž›•‹•ǡ
ƒ ’Š‡‘‡‘ –Šƒ– ™ƒ• ˆ‹”•– ‘„•‡”˜‡† „› ƒ”„—”‰ ȋʹȌǤ • ƒ …‘•‡“—‡…‡ǡ Š‹‰ŠŽ›
’”‘Ž‹ˆ‡”ƒ–‹‰…‡ŽŽ•‡šŠ‹„‹–ƒ‹…”‡ƒ•‡‹–Š‡ƒ‘—–‘ˆŽƒ…–ƒ–‡’”‘†—…–‹‘”‡‰ƒ”†Ž‡••‘ˆ
‘š›‰‡ƒ˜ƒ‹Žƒ„‹Ž‹–›ȋ͵ȌǤ
– ‹• ™‹†‡Ž› ƒ……‡’–‡† –Šƒ– •‡˜‡”ƒŽ ‘…‘‰‡‡•ǡ ‹…Ž—†‹‰ ǡ Ǧͳαǡ ƒ† ȋͶǦ͸Ȍǡ
†”‹˜‡–Š‹••™‹–…Š„›‹…”‡ƒ•‹‰–Š‡‡š’”‡••‹‘ƒ†ƒ…–‹˜‹–›‘ˆ‰Ž›…‘Ž›–‹…‰‡‡•ǡ‹…Ž—†‹‰
‡š‘‹ƒ•‡ ǡ ƒ…–ƒ–‡ ‡Š›†”‘‰‡ƒ•‡ ǡ ›”—˜ƒ–‡ ‹ƒ•‡ ʹǡ ƒ† ›”—˜ƒ–‡
‡Š›†”‘‰‡ƒ•‡‹ƒ•‡ͳȋ͹ǦͻȌǤ
Š‡ ”‡†—…‡† ˆŽ—š –Š”‘—‰Š –Š‡ –”‹…ƒ”„‘š›Ž‹… ƒ…‹† …›…Ž‡ ȋ …›…Ž‡Ȍ ƒ† ‘š‹†ƒ–‹˜‡
’Š‘•’Š‘”›Žƒ–‹‘ ȋȌ ‹• ‘– ƒ……‘’ƒ‹‡† „› …‘’”‘‹•‡† ‹–‘…Š‘†”‹ƒŽ
ˆ—…–‹‘ǡƒ†‹•‘–ƒ••‘…‹ƒ–‡†™‹–ŠƒŽ–‡”ƒ–‹‘•‹–Š‡‡š’”‡••‹‘Ž‡˜‡Ž•‘ˆ‰‡‡•‹˜‘Ž˜‡†
‹‘š‹†ƒ–‹˜‡‡–ƒ„‘Ž‹•Ǥ‡‡š…‡’–‹‘–‘–Š‹•‘–‹‘‹•”‡’”‡•‡–‡†„›–Š‘•‡–—‘”•‹
™Š‹…ŠFHǡSDHǡ‘”IDHƒ”‡—–ƒ–‡†ȋͳͲȌǤ
‘…Šƒ”ƒ…–‡”‹œ‡–Š‡‘Ž‡…—Žƒ”…Šƒ‰‡•”‡•—Ž–‹‰ˆ”‘…‘•–‹–—–‹˜‡ƒ…–‹˜ƒ–‹‘‘ˆ–Š‡͵
’ƒ–Š™ƒ› ‹ –‹••—‡• ™Š‡”‡ –Š‹• ‰‡‡–‹… ƒŽ–‡”ƒ–‹‘ ‹• …ƒ—•ƒŽŽ› Ž‹‡† –‘ ‡‘’Žƒ•–‹…
–”ƒ•ˆ‘”ƒ–‹‘ǡ ™‡ Šƒ˜‡ ‰‡‡”ƒ–‡† ƒ ‘—•‡ ‘†‡Ž ‹ ™Š‹…Š Ž‘•• ‘ˆ –Š‡ Pten –—‘”
•—’’”‡••‘” ‰‡‡ ‹• –ƒ”‰‡–‡† –‘ –Š‡ ˆ‘ŽŽ‹…—Žƒ” ‡’‹–Š‡Ž‹— ‘ˆ –Š‡ –Š›”‘‹† ‰Žƒ† ȋͳͳȌǤ
Š›”‘…›–‡Ǧ•’‡…‹ˆ‹… †‡Ž‡–‹‘ ‘ˆ Pten …‘•–‹–—–‹˜‡Ž› ƒ…–‹˜ƒ–‡• –Š‡ ͵ •‹‰ƒŽ‹‰ …ƒ•…ƒ†‡ǡ
Ž‡ƒ†‹‰–‘Š›’‡”’Žƒ•–‹…–Š›”‘‹†‰Žƒ†•ƒ–„‹”–Šǡ–‘–Š‡†‡˜‡Ž‘’‡–‘ˆ–Š›”‘‹†ˆ‘ŽŽ‹…—Žƒ”
͵
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ƒ†‡‘ƒ•„›͸Ǧͺ‘–Š•‘ˆƒ‰‡ȋͳͳȌǡƒ†‘ˆ‡–ƒ•–ƒ–‹…ˆ‘ŽŽ‹…—Žƒ”…ƒ”…‹‘ƒ•ƒˆ–‡”‘‡
›‡ƒ” ‘ˆ ƒ‰‡ ȋͳʹȌǤ Š‡ ’”‘‰”‡••‹‘ –‘ ‡‘’Žƒ•–‹… –”ƒ•ˆ‘”ƒ–‹‘ ‹• †”ƒƒ–‹…ƒŽŽ›
ƒ……‡Ž‡”ƒ–‡† „› •‹—Ž–ƒ‡‘—• Cdkn1b †‡Ž‡–‹‘ ȋͳʹȌǡ ƒ…–‹˜ƒ–‹‘ ‘ˆ ”ƒ• ȋͳ͵Ȍǡ ‘” Tp53
†‡Ž‡–‹‘ȋͳͶȌǤ
› ƒƒŽ›œ‹‰ –Š‡ ‘Ž‡…—Žƒ” ƒ† ‡–ƒ„‘Ž‹… ƒŽ–‡”ƒ–‹‘• ˆ‘—† ‹ –Š›”‘‹†• ˆ”‘ ›‘—‰ǡ
–—‘”Ǧˆ”‡‡ Pten–Š›”ǦȀǦ ‹…‡ǡ ™‡ Šƒ˜‡ †‹•…‘˜‡”‡† ƒ ‘˜‡Ž ‡…Šƒ‹• ”‡•’‘•‹„Ž‡ ˆ‘” –Š‡
ƒ…–‹˜‡”‡’”‡••‹‘‘ˆ…›…Ž‡ƒ†‹’”‡Ǧ‡‘’Žƒ•–‹…–Š›”‘…›–‡•ǤŠ‹•’ƒ–Š™ƒ›
‹• ‹†‡’‡†‡– ‘ˆ „‘–Š ’”‘Ž‹ˆ‡”ƒ–‹‘ ƒ† ‘ˆ –Š‡ ‘™ ’ƒ–Š™ƒ›• …Žƒ••‹…ƒŽŽ› ƒ••‘…‹ƒ–‡†
™‹–Š –Š‡ ƒ”„—”‰ ‡ˆˆ‡…–Ǥ ‡ •Š‘™ ‹ ˆƒ…– –Šƒ– ͵ ƒ…–‹˜ƒ–‹‘ ‹†—…‡•ǡ –Š”‘—‰Š –Š‡
‹ƒ…–‹˜ƒ–‹‘ ‘ˆ ǡ ƒ …‘‘”†‹ƒ–‡† ”‡’”‡••‹‘ ‘ˆ –Š‡ ‡š’”‡••‹‘ ‘ˆ …›…Ž‡ ƒ†
”‡•’‹”ƒ–‘”›‰‡‡•ǡ™Š‹…Šˆƒ˜‘”•ƒ‡”‘„‹…‰Ž›…‘Ž›•‹•ƒ––Š‡‡š’‡•‡‘ˆǤ
Ͷ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
MATERIALS AND METHODS
Animals.
Š‡PtenL/L ƒ†Ǧ”‡•–”ƒ‹•Šƒ˜‡„‡‡’”‡˜‹‘—•Ž›†‡•…”‹„‡†ȋͳͳȌǤPDKL/L ‹…‡™‡”‡
‹†Ž› ’”‘˜‹†‡† „› ”Ǥ ƒ”‹‘ Ž‡••‹ ȋͳͷȌǤ ŽŽ •–”ƒ‹• ™‡”‡ „ƒ……”‘••‡† ‹ –Š‡ ͳʹͻ˜
„ƒ…‰”‘—†ˆ‘”ƒ–Ž‡ƒ•––‡‰‡‡”ƒ–‹‘•ǡƒ†Ž‹––‡”ƒ–‡•™‡”‡—•‡†ƒ•…‘–”‘Ž•ǤͲͲͳ
ȋ˜‡”‘Ž‹—•ǡ ‹†Ž› ’”‘˜‹†‡† „› ‘˜ƒ”–‹• •–‹–—–‡• ˆ‘” ‹‘‡†‹…ƒŽ ‡•‡ƒ”…Šǡ ƒ•‡Žǡ
™‹–œ‡”Žƒ†Ȍ ™ƒ• ‰‹˜‡ †ƒ‹Ž› „› ’Ǥ‘Ǥ ‰ƒ˜ƒ‰‡ ƒ– ƒ †‘•‡ ‘ˆ ͳͲ ‰Ȁ‰ „‘†› ™‡‹‰Š– ˆ‘” ʹ
™‡‡•ǡ•–ƒ”–‹‰ƒ–ƒ‰‡Ͷ™‡‡•Ǥȋǡ‘”‘–‘ǡƒƒ†ƒȌ™ƒ•‹Œ‡…–‡†‹Ǥ’Ǥƒ–ͶͲͲ
‰Ȁ‰Ȁ†ƒ›ǡˆ‘”Ͷ™‡‡•ǡ•–ƒ”–‹‰ƒ–ƒ‰‡Ͷ™‡‡•Ǥ
Measurement of Glucose Uptake by PET.
‹Ž†–›’‡ƒ†Pten–Š›”ǦȀǦ‹…‡™‡”‡ˆƒ•–‡†‘˜‡”‹‰Š–„‡ˆ‘”‡ƒ–ƒ‹Ž˜‡‹‹Œ‡…–‹‘‘ˆȏͳͺȐ
ˆŽ—‘”‘†‡‘š›‰Ž—…‘•‡ȋ͵ͲͲμ‹ȌǤ‡Š‘—”ƒˆ–‡”‹Œ‡…–‹‘ǡ‹…‡™‡”‡•—„Œ‡…–‡†–‘’‘•‹–”‘
‡‹••‹‘ –‘‘‰”ƒ’Š› ȋȌ •…ƒ‹‰ ™‹–Š –Š‡ ‘…‘”†‡ ‹…”‘•›•–‡• Ͷ ‹…”‘
…ƒ‡”Ǥ‹ƒŽ•™‡”‡‹ƒ‰‡†™Š‹Ž‡ƒ‡•–Š‡–‹œ‡†„›‹ŠƒŽƒ–‹‘™‹–Š‹•‘ˆŽ—”ƒ‡Ǥƒ‰‡
ƒ…“—‹•‹–‹‘™ƒ•†‘‡—•‹‰–Š‡‹…”‘ƒƒ‰‡”™‹–Š–Š‡†‡†‹…ƒ–‡†•‘ˆ–™ƒ”‡Ǥ
Primary Cultures.
Š›”‘‹† ‰Žƒ†• ™‡”‡ ‹…‡† ƒ† ”‡•—•’‡†‡† ‹ ƒǯ• ͳʹȀͳͲΨ ™‹–Š ͳͲͲ ȀŽ
–›’‡…‘ŽŽƒ‰‡ƒ•‡ȋ‹‰ƒǡ–Ǥ‘—‹•ǡ Ȍƒ†ͳȀŽ†‹•’ƒ•‡ȋ‘…Š‡ǡ†‹ƒƒ’‘Ž‹•ǡȌǤ
œ›ƒ–‹…†‹‰‡•–‹‘™ƒ•…ƒ””‹‡†‘—–ˆ‘”ͻͲ‹ƒ–͵͹ιǤˆ–‡”†‹‰‡•–‹‘ǡˆ‘ŽŽ‹…Ž‡•™‡”‡
•‡‡†‡†‹ƒǯ•ͳʹ…‘–ƒ‹‹‰ͶͲΨ—Ǧ‡”—ȋ‘ŽŽƒ„‘”ƒ–‹˜‡‹‘‡†‹…ƒŽǡ‡†ˆ‘”†ǡ
Ȍǡ ‰Ž›ǦŠ‹•ǦŽ›• ȋͳͲ‰ȀŽǡ ‹‰ƒȌǡ ƒ† •‘ƒ–‘•–ƒ–‹ ȋͳͲ‰ȀŽǡ ‹‰ƒȌ ƒ† ƒŽŽ‘™‡† –‘
•’”‡ƒ†ˆ‘”ʹͶǦ͵͸Š„‡ˆ‘”‡’‡”ˆ‘”‹‰–Š‡‡š’‡”‹‡–•Ǥ
Proliferation Analysis.
ͷ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
–‹Ǧ‹͸͹Ǧ•–ƒ‹‡† –Š›”‘‹† •‡…–‹‘• ™‡”‡ ’Š‘–‘‰”ƒ’Š‡† ƒ– šͶͲͲ ƒ‰‹ˆ‹…ƒ–‹‘ ƒ†
ƒƒŽ›œ‡† —•‹‰ –Š‡ ƒ‰‡ •‘ˆ–™ƒ”‡Ǥ ‡–™‡‡ ͳͷͲͲ ƒ† ͵ͲͲͲ …‡ŽŽ• ’‡” •Ž‹†‡ ™‡”‡
ƒƒŽ›œ‡†Ǥ ‘” ”† ‹…‘”’‘”ƒ–‹‘ ‡š’‡”‹‡–•ǡ ‹…‡ ™‡”‡ ‹Œ‡…–‡† ‹Ǥ’Ǥ ™‹–Š ”† ȋͳͲ
‰Ȁ‰ǡ ‹‰ƒȌ †‹••‘Ž˜‡† ‹ ʹ Š‘—”• „‡ˆ‘”‡ •ƒ…”‹ˆ‹…‡Ǥ –‹Ǧ”†Ǧ•–ƒ‹‡† •‡…–‹‘•
™‡”‡ƒƒŽ›œ‡†ƒ•ƒ„‘˜‡Ǥ
Lactate Assay.
ƒ…–ƒ–‡ Ž‡˜‡Ž• ™‡”‡ ƒ••ƒ›‡† —•‹‰ ƒ …‘‡”…‹ƒŽŽ› ƒ˜ƒ‹Žƒ„Ž‡ ‹– ȋ‹‘˜‹•‹‘ǡ ‘—–ƒ‹
‹‡™ǡȌǤƒ…–ƒ–‡Ž‡˜‡Ž•™‡”‡‘”ƒŽ‹œ‡†–‘–Š‡ƒ‘—–‘ˆ ‘”’”‘–‡‹•‡š–”ƒ…–‡†
ˆ”‘‡ƒ…Š–‹••—‡ˆ”ƒ‰‡–‘”’”‹ƒ”›…—Ž–—”‡Ǥ
Oxygen consumption rate and Extracellular acidification rate.
”‡•ŠŽ› ‹•‘Žƒ–‡† –Š›”‘‹† ˆ‘ŽŽ‹…Ž‡• ˆ”‘ ’‘‘Ž• ‘ˆ ͳʹ ™‹Ž† –›’‡ ‘” ͵ —–ƒ– ‹…‡ ™‡”‡
’Žƒ–‡† ƒ• †‡•…”‹„‡† ‹ Dz”‹ƒ”› …—Ž–—”‡•dzǡ ƒ† ‘š›‰‡ …‘•—’–‹‘ ”ƒ–‡ ȋȌ ™ƒ•
‡ƒ•—”‡† —•‹‰ –Š‡ ‡ƒŠ‘”•‡ ʹͶ ‹•–”—‡– ȋ‡ƒŠ‘”•‡ ‹‘•…‹‡…‡•Ȍ —†‡” „ƒ•ƒŽ
…‘†‹–‹‘•ǡ ‹ –Š‡ ’”‡•‡…‡ ‘ˆ –Š‡ ‹–‘…Š‘†”‹ƒŽ ‹Š‹„‹–‘” ‘Ž‹‰‘›…‹ ȋͳ ρ‰ȀŽȌǡ –Š‡
‹–‘…Š‘†”‹ƒŽ—…‘—’Ž‡”ȋͳρȌƒ†–Š‡”‡•’‹”ƒ–‘”›…Šƒ‹‹Š‹„‹–‘”•ƒ–‹›…‹
ȋʹȌƒ†”‘–‡‘‡ȋͲǤͳρȌǤ
Flow cytometry analysis of mitochondrial membrane potential.
”‹ƒ”› –Š›”‘‹† …‡ŽŽ• ™‡”‡ –”‡ƒ–‡† ™‹–Š •‘Ž˜‡– ‘” ʹͲ μ ƒ”„‘›Ž …›ƒ‹†‡ ͶǦ
ȋ–”‹ˆŽ—‘”‘‡–Š‘š›Ȍ’Š‡›ŽŠ›†”ƒœ‘‡ (Ȍ ˆ‘” ͳͲ ‹ ’”‹‘” –‘ •–ƒ‹‹‰ ™‹–Š ͳͲͲ 
‡–”ƒ‡–Š›Ž”Š‘†ƒ‹‡ǡ‡–Š›Ž‡•–‡”ȋǡ˜‹–”‘‰‡Ȍˆ‘”ʹͲ‹Ǥ‡ŽŽ•™‡”‡”‹•‡†‹
ǡ–”›’•‹‹œ‡†ǡƒ†ƒƒŽ›œ‡†ˆ‘”•–ƒ‹‹‰„›ˆŽ‘™…›–‘‡–”›Ǥ
Western Blot Analysis.
͸
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Š›”‘‹†• ƒ† …‡ŽŽ• ™‡”‡ Š‘‘‰‡‹œ‡† ‘ ‹…‡ ‹ „—ˆˆ‡” •—’’Ž‡‡–‡† ™‹–Š
‘’Ž‡–‡’”‘–‡ƒ•‡‹Š‹„‹–‘”–ƒ„Ž‡–ȋ‘…Š‡‹ƒ‰‘•–‹…•ǡ†‹ƒƒ’‘Ž‹•ǡȌǤ‡•–‡”„Ž‘–
ƒƒŽ›•‹• ™ƒ• …ƒ””‹‡† ‘—– —•‹‰ ʹͲǦͶͲ μ‰ ’”‘–‡‹• —•‹‰ ƒ–‹„‘†‹‡• ˆ”‘ ‡ŽŽ ‹‰ƒŽ‹‰
‡…Š‘Ž‘‰›ȋƒ˜‡”•ǡȌǡ‡š…‡’–ˆ‘”…‘‹–ƒ•‡ʹǡ•‘…‹–”ƒ–‡†‡Š›†”‘‰‡ƒ•‡͵ƒǡͳαǡ
ƒ†—……‹ƒ–‡†‡Š›†”‘‰‡ƒ•‡„ȋƒ–ƒ”—œ‹‘–‡…Š‘Ž‘‰›ǡƒ–ƒ”—œǡȌƒ†βǦƒ…–‹
ȋ‹‰ƒǦŽ†”‹…Šǡ–Ǥ‘—‹•ǡȌǤ
Cell Lines and Drug Treatments.
Ǧͳ͵͵ …‡ŽŽ• ȋ‹†Ž› †‘ƒ–‡† „› ”Ǥ ƒ––Š‡™ ‹‰‡Žǡ Š‹‘ –ƒ–‡ ‹˜‡”•‹–›Ȍ ™‡”‡
ƒ‹–ƒ‹‡† ‹ —Ž„‡……‘̵• ‘†‹ˆ‹‡† ƒ‰Ž‡̵• ‡†‹— ȋȌ ™‹–Š ͳͲΨ ˆ‡–ƒŽ „‘˜‹‡
•‡”— ȋȌ ƒ– ͵͹ι ‹ ͷΨ ʹǤ ͺͷͲͷ… …‡ŽŽ• ȋ‹†Ž› †‘ƒ–‡† „› ”Ǥ ƒ”‡Š ƒ”ƒ‰‹ǡ
ƒ••ƒ…Š—•‡––• ‡‡”ƒŽ ‘•’‹–ƒŽȌ ƒ† ͳ͸ …‡ŽŽ• ȋ‹†Ž› †‘ƒ–‡† „› ”Ǥ ‘Š Ǥ
‘’Žƒ†ǡƒ›‘Ž‹‹…Ȍ™‡”‡‰”‘™‹‡†‹—™‹–ŠͳͲΨǤŽŽ…‡ŽŽŽ‹‡•™‡”‡
˜ƒŽ‹†ƒ–‡† ’”‹‘” –‘ •–ƒ”–‹‰ –Š‡ ‡š’‡”‹‡–• „› ƒ’Ž‹ˆ›‹‰ ƒ† •‡“—‡…‹‰ ‰‡‘‹…
ˆ”ƒ‰‡–• ‡…‘’ƒ••‹‰ –Š‡‹” ‘™ —–ƒ–‹‘• ȋͳ͵͵ǣ PTEN ͳ͵ͲȗǡͺͷͲͷ…ǣ BRAF
͸ͲͲǡͳ͸ǣPIK3CAͷͶͷȌǤ
Šƒ”ƒ…‘Ž‘‰‹…ƒŽ ‹Š‹„‹–‘”• ‘ˆ ȋͺͻǡ ‡ŽŽ ‹‰ƒŽ‹‰ ‡…Š‘Ž‘‰‹‡•Ȍǡ ƒ† ͵
ȋͳʹͲǡ‡ŽŽ‡…Š‡‹…ƒŽ•ǡ‘•–‘ǡȌ‘”ƒƒŽ‘‰ȋǡȌ™‡”‡ƒ††‡†ʹͶŠ
ƒˆ–‡”’Žƒ–‹‰Ǥˆ–‡”͵Ͳ‹ǡ•ƒ’Ž‡•™‡”‡…‘ŽŽ‡…–‡†ƒ†’”‡’ƒ”‡†ˆ‘”‡•–‡”„Ž‘–Ǥ‘”
Ž—…‹ˆ‡”ƒ•‡ƒ†“‡š’‡”‹‡–•ǡ•ƒ’Ž‡•™‡”‡…‘ŽŽ‡…–‡†ƒˆ–‡”͹ʹŠǤ
Real Time PCR.
‘–ƒŽ™ƒ•‡š–”ƒ…–‡†™‹–Š”‹œ‘Žƒ†”‡˜‡”•‡–”ƒ•…”‹„‡†—•‹‰–Š‡Š‡”‘•…”‹’–‹–
ȋ‹ˆ‡ ‡…Š‘Ž‘‰‹‡•ǡ ƒ”Ž•„ƒ†ǡ ƒŽ‹ˆ‘”‹ƒȌǤ “Ǧ ™ƒ• ’‡”ˆ‘”‡† ‘ ƒ –‡’‡ Ž—•
ƒ’’ƒ”ƒ–—•—•‹‰–Š‡„•‘Ž—–‡Ž—‡“‘š‹šȋŠ‡”‘…‹‡–‹ˆ‹…ǡƒŽ–ŠƒǡȌƒ†
͹
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ƒ“ƒ‡š’”‡••‹‘ƒ••ƒ›•ȋ’’Ž‹‡†‹‘•›•–‡•ǡƒ”Ž•„ƒ†ǡȌǤƒ…Š•ƒ’Ž‡™ƒ•”—‹
–”‹’Ž‹…ƒ–‡ ƒ† GusB ‘” -actin ™ƒ• —•‡† –‘ …‘–”‘Ž ˆ‘” ‹’—– Ǥ ƒ–ƒ ƒƒŽ›•‹• ™ƒ•
„ƒ•‡† ‘ –Š‡ – ‡–Š‘†ǡ ƒ† ‡š’‡”‹‡–• ™‡”‡ ”‡’‡ƒ–‡† ƒ– Ž‡ƒ•– –Š”‡‡ –‹‡• —•‹‰ ƒ–
Ž‡ƒ•––™‘‹†‡’‡†‡––Š›”‘‹†’‘‘Ž•ȋƒ–Ž‡ƒ•–ˆ‹˜‡‹…‡Ȁ’‘‘ŽȌǤ
AMP, ADP, ATP level determination.
ǡ ǡ ƒ† Ž‡˜‡Ž• ™‡”‡ ƒ••ƒ›‡† —•‹‰ –Š”‡‡ ‹†‡’‡†‡– –Š›”‘‹† ’‘‘Ž• ȋ–‡
‹…‡Ȁ’‘‘ŽȌǤ’’”‘š‹ƒ–‡Ž›͵Ͳ‰‘ˆ–‹••—‡™ƒ•‡š–”ƒ…–‡†‹ʹͳͲρŽ‘ˆ‡š–”ƒ…–‹‘•‘Ž˜‡–ǡ
ͶͲΨȀͶͲΨȀʹͲΨ …‡–‘‹–”‹Ž‡Ȁ‡–Šƒ‘ŽȀͲǤͳΨ ‹ ™ƒ–‡” …‘–ƒ‹‹‰ ƒ ͳͷ̴
‡š–”ƒ…–‹‘•–ƒ†ƒ”†ǤŠ‡…‘Ž—ȏ‡“—‡–Ǧ…ȋ͵Ǥͷ—ǡͳͲͲʹǤͳ‹Ǥ†ǤȌȐ
™ƒ•”—™‹–Šƒ‰”ƒ†‹‡–‘ˆͻͲΨƒ…‡–‘‹–”‹Ž‡ȀͳͲΨ™ƒ–‡”…‘–ƒ‹‹‰ͳͲ‘‹—
‘”ƒ–‡ǡ’͵ǤͲ–‘ͳͲ‘‹—‘”ƒ–‡ǡ’͵ǤͲǡƒ–ͲǤʹŽȀ‹—•‹‰ƒ“—‹–›
ƒ––ƒ…Š‡†–‘ƒƒ–‡”•‡˜‘–”‹’Ž‡“—ƒ†”—’Ž‡Ǥ
Transient transfection assays.
‡ŽŽ•™‡”‡–”ƒ•ˆ‡…–‡†™‹–Šͳρ‰
Ǧͳα‘”ͳ͹͹Ȁͷ͵ͺ’Žƒ•‹†ȋ͓ͳͲʹ͸ǡ͓ͳͺͲͻ͵ǡ
††‰‡‡ǡƒ„”‹†‰‡ǡȌǤˆ–‡”ʹͶŠǡ…‡ŽŽ•™‡”‡–”‡ƒ–‡†™‹–ŠͳȋȌˆ‘”ͶͺŠ
ƒ†…‘ŽŽ‡…–‡†ˆ‘”‹•‘Žƒ–‹‘Ǥ
Dual Luciferase Assay.
‡ŽŽ•™‡”‡…‘Ǧ–”ƒ•ˆ‡…–‡†™‹–ŠͳͲ‰’Š
Ǧ’Žƒ•‹†ȋ”‘‡‰ƒǡƒ†‹•‘ǡȌƒ†ʹρ‰
Ǧͳα’”‘‘–‡”Ž—…‹ˆ‡”ƒ•‡’Žƒ•‹†ȋ͓ͺͺͺ͹ǡ††‰‡‡ǡƒ„”‹†‰‡ǡȌǤˆ–‡”ʹͶŠǡ…‡ŽŽ•
™‡”‡ –”‡ƒ–‡† ™‹–Š ‹Š‹„‹–‘”• ˆ‘” ͶͺŠ ƒ† …‘ŽŽ‡…–‡† ˆ‘” †—ƒŽǦŽ—…‹ˆ‡”ƒ•‡ ”‡’‘”–‡” ƒ••ƒ›
ȋ”‘‡‰ƒȌǤ
Transmission Electron Microscopy.
ͺ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Š›”‘‹†‰Žƒ†•™‡”‡ˆ‹š‡†™‹–ŠʹΨ’ƒ”ƒˆ‘”ƒŽ†‡Š›†‡ƒ†ʹǤͷΨ‰Ž—–ƒ”ƒŽ†‡Š›†‡‹ͲǤͳ
•‘†‹—…ƒ…‘†›Žƒ–‡„—ˆˆ‡”ǡ’‘•–ˆ‹š‡†™‹–ŠͳΨ‘•‹—–‡–”‘š‹†‡ˆ‘ŽŽ‘™‡†„›ͳΨ—”ƒ›Ž
ƒ…‡–ƒ–‡ǡ †‡Š›†”ƒ–‡† –Š”‘—‰Š ƒ ‰”ƒ†‡† •‡”‹‡• ‘ˆ ‡–Šƒ‘Ž ƒ† ‡„‡††‡† ‹ ͳͳʹ ”‡•‹
ȋ ‡•‡ƒ”…Š †—•–”‹‡•ǡ —”Ž‹‰–‘ ȌǤ Ž–”ƒ–Š‹ ȋͺͲ Ȍ •‡…–‹‘• ™‡”‡ …—– ‘ ƒ
‡‹…Š‡”–Ž–”ƒ…—–ǡ•–ƒ‹‡†™‹–Š—”ƒ›Žƒ…‡–ƒ–‡ˆ‘ŽŽ‘™‡†„›Ž‡ƒ†…‹–”ƒ–‡ǡƒ†˜‹‡™‡†
‘ƒͳʹͲͲ–”ƒ•‹••‹‘‡Ž‡…–”‘‹…”‘•…‘’‡ƒ–ͺͲ˜Ǥ
Proteomic Analysis.
‡–• ‘ˆ –Š›”‘‹† ‡š–”ƒ…–• ȋͳͲͲρ‰Ȍ ˆ”‘ ͵Ǧ‘–Š ‘Ž† ™‹Ž† –›’‡ ƒ† —–ƒ– ‹…‡ ™‡”‡
•—„Œ‡…–‡†–‘ʹ‰‡Ž‡Ž‡…–”‘’Š‘”‡•‹•ȋ’ͶǦͳͳȌƒ†›’”‘—„›‰‡Ž•–ƒ‹‹‰ƒ•†‡•…”‹„‡†
‹ȋͳ͸ȌǤ„‘—––Š‹”–›†‹ˆˆ‡”‡–‹ƒŽŽ›‡š’”‡••‡†•’‘–•™‡”‡•‡Ž‡…–‡†ˆ‘”…—––‹‰ƒ†‹Ǧ‰‡Ž
–”›’–‹…†‹‰‡•–‹‘ǤΫ’‡’–‹†‡ƒ••ˆ‹‰‡”’”‹–‹‰™ƒ•’‡”ˆ‘”‡†ƒ•†‡•…”‹„‡†
‹ ȋͳ͸ȌǤ ȀȀ ’‡’–‹†‡ •‡“—‡…‹‰ ™ƒ• ’‡”ˆ‘”‡† ƒ• †‡•…”‹„‡† ‹ ȋͳ͹ȌǤ ŽŽ
ƒƒŽ›•‡•™‡”‡…‘†—…–‡†„›–Š‡‘šŠƒ•‡ƒ…‡”‡–‡””‘–‡‘‹…•ƒ…‹Ž‹–›Ǥ
Statistical Analysis.
š’‡”‹‡–•™‡”‡’‡”ˆ‘”‡†ƒ–Ž‡ƒ•––Š”‡‡–‹‡•Ǥƒ–ƒ™‡”‡ƒƒŽ›œ‡†—•‹‰–Š‡”‹•
•‘ˆ–™ƒ”‡ ’ƒ…ƒ‰‡Ǥ ‹ˆˆ‡”‡…‡• ™‹–Š Ǧ˜ƒŽ—‡• δͲǤͲͷ ™‡”‡ …‘•‹†‡”‡† •–ƒ–‹•–‹…ƒŽŽ›
•‹‰‹ˆ‹…ƒ–Ǥ
RESULTS
Constitutive PI3K activity results in transcriptional repression of TCA cycle and OXPHOS
genes.
Š‡ –Š›”‘‹†• ‘ˆ Pten–Š›”ǦȀǦ ‹…‡ ƒ”‡ ‡Žƒ”‰‡† ˆ”‘ „‹”–Š †—‡ –‘ ƒ ‹…”‡ƒ•‡ ‹ –Š‡
–Š›”‘…›–‡•ǯ ’”‘Ž‹ˆ‡”ƒ–‹˜‡ ‹†‡š ƒ••‘…‹ƒ–‡† ™‹–Š …‘•–‹–—–‹˜‡ ͵ •‹‰ƒŽ‹‰Ǥ ‘™‡˜‡”ǡ
ͻ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
–Š‡•‡ ‰Žƒ†• †‘ ‘– †‹•’Žƒ› ƒ› ˆ—…–‹‘ƒŽ ƒŽ–‡”ƒ–‹‘ ‘” •‹‰• ‘ˆ ‡‘’Žƒ•–‹…
–”ƒ•ˆ‘”ƒ–‹‘—–‹Ž–Š‡‹…‡ƒ”‡ƒ–Ž‡ƒ•–ͳͲ‘–Š•‘ˆƒ‰‡ȋͳʹȌǤ
‘‹†‡–‹ˆ›‡ƒ”Ž›‘Ž‡…—Žƒ”…Šƒ‰‡•–Šƒ–ƒ›ˆƒ˜‘”‘”…‘–”‹„—–‡–‘–Š‡†‡˜‡Ž‘’‡–‘ˆ
–Š›”‘‹†–—‘”•‹ƒ‰‹‰‹…‡ǡ™‡Šƒ˜‡‹‹–‹ƒŽŽ›—•‡†ƒ•ƒŽŽǦ•…ƒŽ‡’”‘–‡‘‹…ƒ’’”‘ƒ…Š–‘
†‡–‡…– ’”‘–‡‹• †‹ˆˆ‡”‡–‹ƒŽŽ› ‡š’”‡••‡† ‹ –Š›”‘‹†• ˆ”‘ ͵ ‘–ŠǦ‘Ž† ™‹Ž† –›’‡ ƒ†
Pten–Š›”ǦȀǦ‹…‡Ǥ‡˜‡”ƒŽ•’‘–•ǡ™‹–Š•‹‰‹ˆ‹…ƒ–Ž›†‹ˆˆ‡”‡–‹–‡•‹–›„‡–™‡‡‘”ƒŽƒ†
—–ƒ––Š›”‘‹†•ǡ™‡”‡•‡Ž‡…–‡†ƒ†‡š…‹•‡†ˆ”‘ʹ‰‡Ž•ǡƒ†–Š‹”–›Ǧˆ‹˜‡’”‘–‡‹•™‡”‡
‹†‡–‹ˆ‹‡† —•‹‰ Ǧ ƒ† ȀȀǤ —”’”‹•‹‰Ž›ǡ •‡˜‡ ‘ˆ –™‡–›Ǧ‡‹‰Š–
’”‘–‡‹• ˆ‘—† †‘™Ǧ”‡‰—Žƒ–‡† ‹ —–ƒ– –Š›”‘‹†• ™‡”‡ ‡œ›‡• ‹˜‘Ž˜‡† ‹ –Š‡ …›…Ž‡ȋƒ„Ž‡ͳȌǤ‘”†‡”–‘˜ƒŽ‹†ƒ–‡–Š‡•‡ˆ‹†‹‰•ǡ™‡‡ƒ•—”‡†„›”‡ƒŽ–‹‡–Š‡
Ž‡˜‡Ž•‘ˆ–Š‡‰‡‡•‡…‘†‹‰–Š‡•‡’”‘–‡‹•ǡƒ†ˆ‘—†–Šƒ–Csǡ Aco2ǡ Idh3aǡ Dldǡ
Pdhbǡ ƒ† SdhA ‡š’”‡••‹‘ ‹ —–ƒ– –Š›”‘‹†• ™ƒ• ”‡’”‘†—…‹„Ž› ʹͲǦͷͲΨ Ž‘™‡”
…‘’ƒ”‡† –‘ –Šƒ– ‘ˆ ™‹Ž† –›’‡ ‰Žƒ†•ǡ ™Š‹Ž‡ –Š‡ ‡š’”‡••‹‘ ‘ˆ –Š‡ ‰Ž—…‘•‡ –”ƒ•’‘”–‡”
Glut1 ™ƒ• •Ž‹‰Š–Ž› ‹…”‡ƒ•‡† ȋ‹‰—”‡ ͳȌǤ ‘™Ǧ”‡‰—Žƒ–‹‘ ‘ˆ –Š‡•‡ ‰‡‡• ™ƒ• ƒŽ•‘
…‘ˆ‹”‡† „› ‡•–‡” „Ž‘––‹‰ ƒƒŽ›•‹• —•‹‰ ƒ–‹„‘†‹‡• •’‡…‹ˆ‹… ˆ‘” …‘‹–ƒ•‡ ʹǡ
•‘…‹–”ƒ–‡†‡Š›†”‘‰‡ƒ•‡͵ƒǡƒ†—……‹ƒ–‡†‡Š›†”‘‰‡ƒ•‡„ȋ‹‰—”‡ͳȌǤ
‘ †‡–‡”‹‡ –‘ ™Šƒ– ‡š–‡– ‡–ƒ„‘Ž‹… ‰‡‡• ƒ”‡ –”ƒ•…”‹’–‹‘ƒŽŽ› †‡”‡‰—Žƒ–‡† —’‘
͵ƒ…–‹˜ƒ–‹‘ǡ™‡‹–‡””‘‰ƒ–‡†‘—”ˆˆ›‡–”‹š‰‡‡‡š’”‡••‹‘†ƒ–ƒ•‡–ȋͳͶȌ–‘‡š–‡†
–Š‡ ƒƒŽ›•‹• –‘ ƒŽŽ –Š‡ ‰‡‡• †‹”‡…–Ž› ‹˜‘Ž˜‡† ‹ –Š‡ …›…Ž‡ ƒ† ‰Ž›…‘Ž›•‹•Ǥ
–‡”‡•–‹‰Ž›ǡ™Š‹Ž‡‘‡‘ˆ–Š‡‰Ž›…‘Ž›–‹…‡œ›‡•™ƒ•–”ƒ•…”‹’–‹‘ƒŽŽ›†‡”‡‰—Žƒ–‡†‹
–Š‡ —–ƒ– ‰Žƒ†• ȋ™‹–Š –Š‡ ‡š…‡’–‹‘ ‘ˆ ƒ ‘†‡•– ‹…”‡ƒ•‡ ‹ –Š‡ Glucokinase 
Ž‡˜‡Ž•Ȍǡ–Š‡‡š’”‡••‹‘‘ˆʹʹ‘—–‘ˆʹͺ‰‡‡•‡…‘†‹‰‡œ›‡•ƒ••‘…‹ƒ–‡†™‹–Š–Š‡
…›…Ž‡ ™ƒ• •‹‰‹ˆ‹…ƒ–Ž› ”‡’”‡••‡† ‹ PtenǦȀǦ –Š›”‘‹†• ȋ‹‰—”‡ ͳ ƒ† †ƒ–ƒ ‘– •Š‘™ȌǤ
ͳͲ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
„‘—– ͸ͲΨ ‘ˆ –Š‡•‡ ‰‡‡• ™‡”‡ •–‹ŽŽ ”‡’”‡••‡† ‹ –Š‡ ‡–ƒ•–ƒ–‹… ˆ‘ŽŽ‹…—Žƒ” …ƒ”…‹‘ƒ•
†‡˜‡Ž‘’‹‰‹‘Ž†‡”Pten–Š›”ǦȀǦ‹…‡ȋ‹‰—”‡ͳȌǤ
—”–Š‡”‘”‡ǡ™Š‡™‡“—ƒ–‹–ƒ–‡†…‹–”ƒ–‡ƒ†‹•‘…‹–”ƒ–‡‹–Š‡–Š›”‘‹†•‘ˆ…‘–”‘Žƒ†
Pten–Š›”ǦȀǦ‹…‡„›
Ȁǡ™‡ˆ‘—†•‹‰‹ˆ‹…ƒ–Ž›”‡†—…‡†Ž‡˜‡Ž•‘ˆ–Š‡•‡‡–ƒ„‘Ž‹–‡•‹
–Š‡—–ƒ–‰Žƒ†•ȋ…‹–”ƒ–‡ǣʹǡͷ͹͹άͷʹͷ’‘ŽȀ‰‹–Š‡™–ƒ†ͻͺͷά͵Ͳʹ‹–Š‡—–ƒ–•ǡ
‹•‘…‹–”ƒ–‡ʹͳ͹άͶʹ’‘ŽȀ‰‹–Š‡™–ƒ†ͺͲάʹ͸‹–Š‡—–ƒ–•ȌǤ
Š‡–‹‰Š–…‘‡…–‹‘‡š‹•–‹‰„‡–™‡‡…›…Ž‡ǡ‘š‹†ƒ–‹˜‡’Š‘•’Š‘”›Žƒ–‹‘ȋȌǡ
ƒ† ‹–‘…Š‘†”‹ƒŽ ˆ‹–‡•• ’”‘’–‡† —• –‘ –‡•– ™Š‡–Š‡” –Š‹• ‰Ž‘„ƒŽ †‘™Ǧ”‡‰—Žƒ–‹‘ ‘ˆ
…›…Ž‡ ‰‡‡• ‹• ƒ……‘’ƒ‹‡† „› ƒŽ–‡”ƒ–‹‘• ‘ˆ –Š‡ ‡š’”‡••‹‘ ‘ˆ ‹–‘…Š‘†”‹ƒǦ
‡…‘†‡†‰‡‡•ǡƒ•™‡ŽŽƒ•„›…Šƒ‰‡•‹–Š‡—„‡”‘ˆ‹–‘…Š‘†”‹ƒǤ‡ˆ‹”•–
—•‡† ”‡ƒŽ –‹‡ –‘ ‡ƒ•—”‡ –Š‡ ”‡Žƒ–‹˜‡ ‹–‘…Š‘†”‹ƒŽ ‰‡‘‡ …‘’› —„‡” „›
ƒ’Ž‹ˆ›‹‰ˆ‘—”†‹ˆˆ‡”‡–‹–‘…Š‘†”‹ƒŽŽ›‡…‘†‡†‰‡‡•ƒ†‘‡”‡ˆ‡”‡…‡—…Ž‡ƒ”‰‡‡
ˆ”‘ –‘–ƒŽ ȋ‰‡‘‹… ƒ† ‹–‘…Š‘†”‹ƒŽȌ ‹•‘Žƒ–‡† ˆ”‘ ™‹Ž† –›’‡ ƒ† —–ƒ–
–Š›”‘‹†•Ǥ ‘ •‹‰‹ˆ‹…ƒ– †‹ˆˆ‡”‡…‡• ™‡”‡ ˆ‘—† „‡–™‡‡ …‘–”‘Ž ƒ† —–ƒ–•ǡ
•—‰‰‡•–‹‰ –Šƒ– …‘•–‹–—–‹˜‡ ͵ ƒ…–‹˜ƒ–‹‘ †‘‡• ‘– ƒŽ–‡” ‹–‘…Š‘†”‹ƒŽ ƒ•• ‹ –Š‡
‘—•‡ –Š›”‘‹† ȋ‹‰—”‡ ͳȌǤ ‘˜‡”•‡Ž›ǡ–Š‡‡š’”‡••‹‘‘ˆ ‘•–‹–‘…Š‘†”‹ƒǦ‡…‘†‡†
‰‡‡•™ƒ•Š‡ƒ˜‹Ž›†‘™Ǧ”‡‰—Žƒ–‡†‹PtenǦȀǦ–Š›”‘‹†•ȋ‹‰—”‡ͳȌǤ—”–Š‡”‘”‡ǡ‡Ž‡…–”‘
‹…”‘•…‘’› ƒƒŽ›•‹• ‘ˆ …‘–”‘Ž ƒ† —–ƒ– –Š›”‘‹† •‡…–‹‘• •Š‘™‡† –Šƒ– Pten Ž‘••
…ƒ—•‡†’”‘ˆ‘—†‘”’Š‘Ž‘‰‹…ƒŽ†‡ˆ‡…–•‹–Š‡‹–‘…Š‘†”‹ƒǡ‹…Ž—†‹‰•™‡ŽŽ‹‰ǡƒ–”‹š
…Žƒ”‹ˆ‹…ƒ–‹‘ǡƒ††‹•”—’–‹‘‘ˆ–Š‡…”‹•–ƒ‡ȋ‹‰—”‡ͳȌǤŽŽ–Š‡•‡ˆ‡ƒ–—”‡•ƒ”‡…‘‘Ž›
ƒ••‘…‹ƒ–‡†™‹–Š”‡†—…‡†‹–‘…Š‘†”‹ƒŽ‘š‹†ƒ–‹˜‡…ƒ’ƒ…‹–›ȋͳͺȌǤ
‘ –‡•– ™Š‡–Š‡” –Š‡ ƒˆ‘”‡‡–‹‘‡† ˆ‹†‹‰• ”‡ˆŽ‡…– ƒ ‹’ƒ‹”‡– ‹ ‹–‘…Š‘†”‹ƒŽ
ˆ—…–‹‘ǡ ™‡ ‡ƒ•—”‡† ‘š›‰‡ …‘•—’–‹‘ ”ƒ–‡• ȋȌ ƒ• ™‡ŽŽ ƒ• ‡š–”ƒ…‡ŽŽ—Žƒ”
ͳͳ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ƒ…‹†‹ˆ‹…ƒ–‹‘ ”ƒ–‡ ȋȌ ‹ ’”‹ƒ”› …—Ž–—”‡• ‘ˆ …‘–”‘Ž ƒ† PtenǦȀǦ –Š›”‘…›–‡•Ǥ
–”‹‹‰Ž›ǡPtenǦȀǦ–Š›”‘…›–‡•‡šŠ‹„‹–‡†•‹‰‹ˆ‹…ƒ–Ž›”‡†—…‡†•’ƒ”‡”‡•’‹”ƒ–‘”›…ƒ’ƒ…‹–›ǡ
‹Ǥ‡Ǥ Ž‘™‡” ƒš‹ƒŽ ”ƒ–‡ ‘ˆ ”‡•’‹”ƒ–‹‘ ȋ‹‰—”‡ ͳ
Ȍ ƒ† ‹…”‡ƒ•‡† ȋ‹‰—”‡ ͳȌǤ
‹ƒŽŽ›ǡ ™‡ —•‡† ˆŽ‘™ …›–‘‡–”› –‘ …‘’ƒ”‡ –Š‡ ‹–‘…Š‘†”‹ƒŽ ‡„”ƒ‡ ’‘–‡–‹ƒŽ ‹
’”‹ƒ”›…—Ž–—”‡•‘ˆ…‘–”‘Žƒ†PtenǦȀǦ–Š›”‘…›–‡•—•‹‰–Š‡…ƒ–‹‘‹…’‘–‡–‹ƒŽǦ•‡•‹–‹˜‡
†›‡ǡ –‡–”ƒ‡–Š›Ž”Š‘†ƒ‹‡Ǧ‡–Š›ŽǦ‡•–‡” ȋȌǡ ™Š‹…Š ƒ……——Žƒ–‡• ‹ ‹–ƒ…–
‹–‘…Š‘†”‹ƒǤ Š‡ ƒ‹ ’‘’—Žƒ–‹‘ ‘ˆ —–ƒ– …‡ŽŽ• †‹•’Žƒ›‡† ”‡†—…‡† ‹–‘…Š‘†”‹ƒŽ
‡„”ƒ‡ ’‘–‡–‹ƒŽ …‘’ƒ”‡† –‘ …‘–”‘Ž –Š›”‘…›–‡• ȋƒ„‘—– ͷͲΨ ‘ˆ ™‹Ž† –›’‡ Ž‡˜‡Ž•ǡ
‘–‡ –Š‡ Ž‘‰ƒ”‹–Š‹… •…ƒŽ‡ǡ ‹‰—”‡ ͳȌǡ ™Š‹Ž‡ ƒ •‡…‘† ’‘’—Žƒ–‹‘ ‘ˆ —–ƒ– …‡ŽŽ• ™ƒ•
…Šƒ”ƒ…–‡”‹œ‡†„›‡˜‡Ž‘™‡”’‘–‡–‹ƒŽǡ•‹‹Žƒ”–‘–Šƒ–‘ˆ…‡ŽŽ•–”‡ƒ–‡†™‹–Š–Š‡—…‘—’Ž‡”
Ǥ
ƒ‡–‘‰‡–Š‡”ǡ–Š‡•‡†ƒ–ƒ†‡‘•–”ƒ–‡–Šƒ–…‘•–‹–—–‹˜‡͵ƒ…–‹˜ƒ–‹‘‹–Š‡‘—•‡
–Š›”‘‹†”‡•—Ž–•‹–Š‡…‘‘”†‹ƒ–‡††‘™Ǧ”‡‰—Žƒ–‹‘‘ˆ–Š‡‡š’”‡••‹‘‘ˆ‰‡‡•‡…‘†‹‰
‡„‡”• ‘ˆ „‘–Š –Š‡ …›…Ž‡ ƒ† ’ƒ–Š™ƒ›•ǡ ‹ ‡š–‡•‹˜‡ ‹–‘…Š‘†”‹ƒŽ
†ƒƒ‰‡ǡƒ†‹•‡˜‡”‡‹’ƒ‹”‡–‘ˆ‹–‘…Š‘†”‹ƒŽˆ—…–‹‘Ǥ
A glycolytic switch in pre-neoplastic Pten-/- thyrocytes.
‹…‡ –Š‡ ‡š’”‡••‹‘ …Šƒ‰‡• †‡•…”‹„‡† ƒ„‘˜‡ ”‡•—Ž– ‹ ”‡†—…‡† ‡ˆˆ‹…‹‡…› ‘ˆ –Š‡ …›…Ž‡Ȁ ‡–ƒ„‘Ž‹… ’ƒ–Š™ƒ›ǡ –Š‡ —–ƒ– …‡ŽŽ• ‹‰Š– Šƒ˜‡ –‘ ‹…”‡ƒ•‡ –Š‡‹”
‰Ž›…‘Ž›–‹…”ƒ–‡–‘‡‡––Š‡‹”‡‡”‰‡–‹…‡‡†•Ǥ
•ƒ’”‘š›ˆ‘”–Š‡–Š›”‘…›–‡•ǯ‰Ž›…‘Ž›–‹…”ƒ–‡ǡ™‡ƒ••‡••‡†–Š‡ƒ‘—–‘ˆŽƒ…–ƒ–‡’”‡•‡–
‹ –Š›”‘‹†• ˆ”‘ ͵Ǧ‘–Š ‘Ž† ™‹Ž† –›’‡ ƒ† —–ƒ– ‹…‡Ǥ –”‹‹‰Ž›ǡ PtenǦȀǦ –Š›”‘‹†•
†‹•’Žƒ›‡† ƒ –Š”‡‡Ǧˆ‘Ž† ‹…”‡ƒ•‡ ‹ ‘”ƒŽ‹œ‡† Žƒ…–ƒ–‡ …‘–‡–ǡ •—‰‰‡•–‹‰ ƒ Š‹‰Š‡”
‡–ƒ„‘Ž‹… ˆŽ—š –Š”‘—‰Š ‰Ž›…‘Ž›•‹• ȋ‹‰—”‡ ʹȌǤ Ž‘‰ –Š‡ •ƒ‡ Ž‹‡ǡ ™‡ ‡ƒ•—”‡† –Š‡
ͳʹ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Žƒ…–ƒ–‡•‡…”‡–‹‘”ƒ–‡‹’”‹ƒ”›–Š›”‘…›–‡…—Ž–—”‡•Ǥ—–ƒ–…‡ŽŽ•’”‘†—…‡†͵Ǧ–‘ͷǦˆ‘Ž†
‘”‡ Žƒ…–ƒ–‡ –Šƒ –Š‡‹” ™‹Ž† –›’‡ …‘—–‡”’ƒ”–• ȋ‹‰—”‡ ʹȌǡ •–”‘‰Ž› ‹†‹…ƒ–‹‰ –Šƒ–
‰Ž›…‘Ž›•‹• ‹• ‹†‡‡† —’Ǧ”‡‰—Žƒ–‡† –‘ …‘’‡•ƒ–‡ –Š‡ ͵Ǧ‡†‹ƒ–‡† ”‡’”‡••‹‘ ‘ˆ –Š‡
…›…Ž‡Ȁ’ƒ–Š™ƒ›Ǥ‹ƒŽŽ›ǡ–‘˜ƒŽ‹†ƒ–‡–Š‹•Š›’‘–Š‡•‹•ǡ™‡’‡”ˆ‘”‡† ͳͺ
Ǧ
‹ƒ‰‹‰ ‘ …‘–”‘Ž ƒ† Pten–Š›”ǦȀǦ ‹…‡ǡ ƒ† ˆ‘—† –Šƒ– –Š‡ Š›’‡”’Žƒ•–‹… PtenǦȀǦ
–Š›”‘‹†••Š‘™‡†•–”‘‰ ͳͺ
—’–ƒ‡ǡ™Š‹Ž‡‘—’–ƒ‡™ƒ•†‡–‡…–‡†‹™‹Ž†–›’‡‹…‡
ȋ‹‰—”‡ʹȌǤ
Š‡•‡†ƒ–ƒ•Š‘™–Šƒ–ǡ‹”‡•’‘•‡–‘…‘•–‹–—–‹˜‡͵ƒ…–‹˜ƒ–‹‘ǡ–Š›”‘‹†‡’‹–Š‡Ž‹ƒŽ…‡ŽŽ•
—†‡”‰‘ ƒ ‰Ž›…‘Ž›–‹… •™‹–…Šǡ ™Š‹…Š ‹• ˆ—…–‹‘ƒŽŽ› ”‡‹‹•…‡– ‘ˆ –Š‡ ƒ”„—”‰ ‡ˆˆ‡…–
‘„•‡”˜‡† ‹ …ƒ…‡” …‡ŽŽ•ǡ ƒ• ™‡ŽŽ ƒ• ‹ Š‹‰ŠŽ› ’”‘Ž‹ˆ‡”ƒ–‹‰ …‡ŽŽ• •—…Š ƒ• ƒ…–‹˜ƒ–‡†
Ž›’Š‘…›–‡•Ǥ
Š‡ƒ”„—”‰‡ˆˆ‡…–‹•—•—ƒŽŽ›†”‹˜‡„›ƒ•‡–‘ˆDzƒ•–‡”‰‡‡•dzǡ‹…Ž—†‹‰…Ǧǡǡ
ǡ ƒ† ͳαǡ ™Š‹…Š …‘–”‘Ž –Š‡ ‡š’”‡••‹‘ǡ Ž‘…ƒŽ‹œƒ–‹‘ ƒ† ˆ—…–‹‘ ‘ˆ „‘–Š
‰Ž—…‘•‡ –”ƒ•’‘”–‡”• ƒ† ‰Ž›…‘Ž›–‹… ‡œ›‡• ȋͷǡ ͳͻȌǤ ’Ǧ”‡‰—Žƒ–‹‘ ‘ˆ ‡š‘‹ƒ•‡ ʹǡ
›”—˜ƒ–‡ ‹ƒ•‡ ʹǡ ƒ† ƒ…–ƒ–‡ †‡Š›†”‘‰‡ƒ•‡• ƒ† ƒ”‡ ŠƒŽŽƒ”• ‘ˆ –Š‹•
‰Ž›…‘Ž›–‹…•™‹–…ŠǤ
Š‡‡•–ƒ„Ž‹•Š‡–‘ˆƒ‰Ž›…‘Ž›–‹…•™‹–…Š‹•Ž‘™Ž›’”‘Ž‹ˆ‡”ƒ–‹‰ǡ‘Ǧ‡‘’Žƒ•–‹…ȋ‹Ǥ‡Ǥ‘Ǧ
–”ƒ•ˆ‘”‡†Ȍ…‡ŽŽ•ǡƒ†–Š‡ƒ„•‡…‡‘ˆ‰Ž›…‘Ž›–‹…‰‡‡•—’Ǧ”‡‰—Žƒ–‹‘‹‘—”‡š’”‡••‹‘
’”‘ˆ‹Ž‹‰†ƒ–ƒ™‘—Ž†ƒ”‰—‡ƒ‰ƒ‹•–ƒ‡…Šƒ‹•‹˜‘Ž˜‹‰–Š‡…Žƒ••‹…ƒŽƒ”„—”‰‡ˆˆ‡…–Ǧ
”‡Žƒ–‡† ƒ•–‡” ”‡‰—Žƒ–‘”•Ǥ  ˆƒ…–ǡ ™‡ †‹† ‘– †‡–‡…– •‹‰‹ˆ‹…ƒ– ƒŽ–‡”ƒ–‹‘• ‹ –Š‡
‡š’”‡••‹‘Ž‡˜‡Ž•‘ˆHif1αǡc-MycǡHk2ǡPkm2ǡLdhaǡ‘”Ldhb„‘–Šƒ––Š‡ƒ†’”‘–‡‹
Ž‡˜‡Žȋ‹‰—”‡ʹǡƒ††ƒ–ƒ‘–•Š‘™ȌǤ
ͳ͵
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Š‡•‡ †ƒ–ƒ •—‰‰‡•– –Šƒ– ͵ ƒ…–‹˜ƒ–‹‘ ‹ ‘Ǧ–”ƒ•ˆ‘”‡† –Š›”‘‹† ‡’‹–Š‡Ž‹ƒŽ …‡ŽŽ•
‹†—…‡•ƒ‰Ž›…‘Ž›–‹…•™‹–…Š–Š”‘—‰Šƒ‘˜‡Ž‘Ž‡…—Žƒ”‡…Šƒ‹•Ǥ
Pdk1 drives TCA cycle/OXPHOS gene repression downstream of PI3K, independent of
mTOR.
†ͳ‹•ƒ‡••‡–‹ƒŽ‡„‡”‘ˆ–Š‡͵…ƒ•…ƒ†‡ǡ•‹…‡‹–’Š‘•’Š‘”›Žƒ–‡•–‘͵Ͳͺ
ƒ† ͸ͳ ‘ ʹʹͻǡ ƒŽŽ‘™‹‰ –Š‡‹” ˆ—ŽŽ ƒ…–‹˜ƒ–‹‘ ȋʹͲȌǤ ‘ –‡•– –‘ ™Šƒ– ‡š–‡– –Š‡ …›…Ž‡ ‰‡‡ ”‡’”‡••‹‘ ‘„•‡”˜‡† ‹ PtenǦȀǦ –Š›”‘‹†• †‡’‡†• ‘ –Š‡ †ͳȀ–Ȁ
…ƒ•…ƒ†‡ǡ ™‡ ˆ‹”•– …”‘••‡† Pten–Š›”ǦȀǦ ‹…‡ –‘ Pdk1 …‘†‹–‹‘ƒŽ —–ƒ–• ȋʹͳȌǡ ‘„–ƒ‹‹‰
•‹—Ž–ƒ‡‘—• –Š›”‘‹†Ǧ•’‡…‹ˆ‹… †‡Ž‡–‹‘ ‘ˆ –Š‡•‡ –™‘ ‰‡‡•Ǥ ‘„‹‡† Ž‘•• ‘ˆ Pten ƒ†
Pdk1 ”‡•…—‡† „‘–Š –Š‡ ”‡’”‡••‹‘ ‘ˆ –Š‡ …›…Ž‡ ‰‡‡• ȋ‹‰—”‡ ͵Ȍ ƒ† –Šƒ– ‘ˆ
‹–‘…Š‘†”‹ƒŽŽ›Ǧ‡…‘†‡† ‰‡‡• ȋ‹‰—”‡ ͵ȌǤ –‡”‡•–‹‰Ž›ǡ –Ǧ‡…‘†‡†
‰‡‡ ‡š’”‡••‹‘ ‹ –Š‡ …‘’‘—† —–ƒ–• ™ƒ• Š‹‰Š‡” –Šƒ ‹ ™‹Ž† –›’‡ …‘–”‘Ž•Ǥ
—”–Š‡”‘”‡ǡ•‹—Ž–ƒ‡‘—•‹ƒ…–‹˜ƒ–‹‘‘ˆPtenƒ†Pdk1”‡•–‘”‡†Žƒ…–ƒ–‡’”‘†—…–‹‘‹
–Š‡–Š›”‘‹†–‘™‹Ž†–›’‡Ž‡˜‡Ž•ȋ‹‰—”‡͵ȌǤ
‘•–‹–—–‹˜‡ ͵ ƒ…–‹˜ƒ–‹‘ ‹ –Š‡ –Š›”‘‹† ˆ‘ŽŽ‹…—Žƒ” …‡ŽŽ• ”‡•—Ž–• ‹  ƒ…–‹˜ƒ–‹‘
ȋʹʹȌǡ ƒ†  ƒ…–‹˜‹–› Šƒ• „‡‡ ’”‘’‘•‡† –‘ ’Žƒ› ƒ ‡› ”‘Ž‡ ‹ ‡•–ƒ„Ž‹•Š‹‰ –Š‡
‰Ž›…‘Ž›–‹… •™‹–…Š ȋʹ͵ȌǤ ‘ –‡•– –Š‡ ”‘Ž‡ ‘ˆ –Š‡ ͵Ȁ–Ȁ ƒš‹• ‹ –Š‡ ‡–ƒ„‘Ž‹…
’Š‡‘–›’‡ ‘ˆ Pten–Š›”ǦȀǦ ‹…‡ǡ ™‡ –”‡ƒ–‡† ͵Ǧ‘–Š ‘Ž† ™‹Ž† –›’‡ ƒ† —–ƒ– ‹…‡
ȋαͷȀ‰”‘—’Ȍ†ƒ‹Ž›ˆ‘”ʹ™‡‡•™‹–Š–Š‡‹Š‹„‹–‘”ͲͲͳ‘”’Žƒ…‡„‘ȋ‹‰—”‡͵ȌǤ
•’”‡˜‹‘—•Ž›•Š‘™„›‘—”‰”‘—’ǡ–Š‹•–”‡ƒ–‡–‹••—ˆˆ‹…‹‡––‘‹Š‹„‹–ƒ…–‹˜‹–›
ƒ† †”ƒ•–‹…ƒŽŽ› ”‡†—…‡ ’”‘Ž‹ˆ‡”ƒ–‹‘ ‹ —–ƒ– –Š›”‘‹†• ȋʹʹȌ ȋ‹‰ǡ ͵ǡȌǤ –”‹‹‰Ž›ǡ
†‡•’‹–‡–Š‡…Ž‡ƒ”‹Š‹„‹–‹‘‘ˆ–Š‡–Š›”‘…›–‡•ǯ’”‘Ž‹ˆ‡”ƒ–‹‘”ƒ–‡ǡͲͲͳ–”‡ƒ–‡–Šƒ†
ͳͶ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
‘ ‡ˆˆ‡…– ‘ –Š‡ ”‡’”‡••‹‘ ‘ˆ …›…Ž‡ ‰‡‡• ȋ‹‰—”‡ ͵Ȍǡ ‘ –Š‡ ”‡’”‡••‹‘ ‘ˆ
‹–‘…Š‘†”‹ƒŽ‰‡‡•ȋ‹‰—”‡͵
Ȍǡ‘”‘–Š›”‘‹†Žƒ…–ƒ–‡’”‘†—…–‹‘ȋ‹‰—”‡͵ȌǤ
Š‡•‡ †ƒ–ƒ •–”‘‰Ž› •—‰‰‡•– –Šƒ– ͵ ƒ…–‹˜ƒ–‹‘ ‹ –Š›”‘‹† …‡ŽŽ• ‹†—…‡• ƒ ‰Ž›…‘Ž›–‹…
•™‹–…Š ‹†‡’‡†‡– ‘ˆ  ƒ…–‹˜‹–›Ǥ —”–Š‡”‘”‡ǡ –Š‡› •Š‘™ –Šƒ– –Š‡ ‹…”‡ƒ•‡ ‹
‰Ž›…‘Ž›•‹• ‹ Pten–Š›”ǦȀǦ ‹…‡ ‹• ‘– ƒ †‹”‡…– …‘•‡“—‡…‡ ‘ˆ –Š‡ ‹…”‡ƒ•‡† ’”‘Ž‹ˆ‡”ƒ–‹˜‡
”ƒ–‡‘„•‡”˜‡†‹–Š‡Š›’‡”’Žƒ•–‹…‰Žƒ†•Ǥ
PI3K/AKT inhibit AMPK, leading to decreased OXPHOS metabolism in pre-neoplastic
thyrocytes.
Š‡‡š’”‡••‹‘‘ˆ—…Ž‡—•Ǧƒ†‹–‘…Š‘†”‹ƒǦ‡…‘†‡†”‡•’‹”ƒ–‘”›’”‘–‡‹•‹•…‘–”‘ŽŽ‡†
„› ƒ —„‡” ‘ˆ —…Ž‡—•Ǧ‡…‘†‡† –”ƒ•…”‹’–‹‘ ˆƒ…–‘”• ƒ† …‘Ǧƒ…–‹˜ƒ–‘”• –Šƒ– ‘†—Žƒ–‡
‹–‘…Š‘†”‹ƒŽˆ—…–‹‘‹”‡•’‘•‡–‘‡š–”ƒǦƒ†‹–”ƒ…‡ŽŽ—Žƒ”•‹‰ƒŽ•Ǥ
‘”†‡”–‘‹†‡–‹ˆ›–Š‡‡…Šƒ‹•–Š”‘—‰Š™Š‹…ŠŽ‘••‘ˆPten”‡•—Ž–•‹ƒ…‘‘”†‹ƒ–‡†
”‡’”‘‰”ƒ‹‰‘ˆ–Š‡‡š’”‡••‹‘‘ˆ”‡•’‹”ƒ–‘”›‰‡‡•ǡ™‡‹‹–‹ƒŽŽ›‡ƒ•—”‡†‹–Š›”‘‹†•
ˆ”‘͵‘–ŠǦ‘Ž†™‹Ž†–›’‡ƒ†Pten–Š›”ǦȀǦ‹…‡–Š‡‡š’”‡••‹‘Ž‡˜‡Ž•‘ˆ–Š‡–”ƒ•…”‹’–‹‘
ˆƒ…–‘”• ƒ† …‘ˆƒ…–‘”• ‹˜‘Ž˜‡† ‹ –Š‡ ”‡‰—Žƒ–‹‘ ‘ˆ …›…Ž‡ȀǤ Š‹Ž‡ –Š‡
‡š’”‡••‹‘Ž‡˜‡Ž•‘ˆERRβ, PRC, NRF-1ƒ†-2ǡƒ†TFAM™‡”‡‘–ƒŽ–‡”‡†„›…‘•–‹–—–‹˜‡
͵ ƒ…–‹˜ƒ–‹‘ǡ Pten–Š›”ǦȀǦ –Š›”‘‹†• Šƒ† •‹‰‹ˆ‹…ƒ–Ž› Ž‘™‡” ‡š’”‡••‹‘ ‘ˆ ERRγ, PGC-1α
ƒ† –βǡ ƒ†ǡ –‘ ƒ Ž‡••‡” ‡š–‡–ǡ ERRα ȋ‹‰—”‡ ͶȌǤ Š‡•‡ †ƒ–ƒ •—‰‰‡•– –Šƒ– ͵
ƒ…–‹˜ƒ–‹‘‹‰Š–ƒŽ–‡”–Š‡‡š’”‡••‹‘‘”ƒ…–‹˜‹–›‘ˆƒ…‘‘ƒ•–‡””‡‰—Žƒ–‘”›‰‡‡Ǥ
‡…‡–Ž› ’—„Ž‹•Š‡† †ƒ–ƒ Šƒ˜‡ †‹”‡…–‡† ‘—” ƒ––‡–‹‘ ‘ ƒ• ƒ …ƒ†‹†ƒ–‡ ˆ‘” –Š‹•
”‘Ž‡Ǥ …‘–”‹„—–‡• –‘ –Š‡ …‘–”‘Ž ‘ˆ ”‡•’‹”ƒ–‘”› ‰‡‡• ‹ —•…Ž‡ …‡ŽŽ• ȋʹͶȌ ƒ†
’Š‘•’Š‘”›Žƒ–‡• Ǧͳα ‘ ͳ͹͹ ƒ† ͷ͵ͺ –‘ ‹…”‡ƒ•‡ ‹–• –”ƒ•…”‹’–‹‘ƒŽ ƒ…–‹˜‹–›ǡ
ͳͷ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
‹…Ž—†‹‰ ‹–• ƒ„‹Ž‹–› –‘ –”ƒ•ƒ…–‹˜ƒ–‡ ‹–• ‘™ ’”‘‘–‡” ȋʹͷȌǤ  ƒ††‹–‹‘ǡ –Š‡ ESRRA
’”‘‘–‡” …‘–ƒ‹• ƒ ǦͳαȀαǦ”‡•’‘•‹˜‡ ”‡‰—Žƒ–‘”› ‡Ž‡‡– –Šƒ– …ƒ ƒŽ•‘ „‡
”‡…‘‰‹œ‡† ƒ† ƒ…–‹˜ƒ–‡† „› β ƒ† γ ȋʹ͸ȌǤ —”–Š‡”‘”‡ǡ ‹ …‡”–ƒ‹ …‡ŽŽ –›’‡•ǡ
…ƒ ’Š‘•’Š‘”›Žƒ–‡ ‘ Ͷͺͷǡ ’”‡˜‡–‹‰ ͳ ˆ”‘ ’Š‘•’Š‘”›Žƒ–‹‰ ȋ‘
ͳ͹ʹȌƒ†ƒ…–‹˜ƒ–‹‰ȋʹ͹ǡʹͺȌǤ†‡‡†ǡ™‡ˆ‘—†–Šƒ––Š›”‘‹†•ˆ”‘Pten–Š›”ǦȀǦ‹…‡
Šƒ˜‡ †”ƒ•–‹…ƒŽŽ› ”‡†—…‡† ’ͳ͹ʹǦ ƒ† Š‹‰Š‡” ’ͶͺͷǦǡ …‘’ƒ”‡† –‘ ™‹Ž† –›’‡
…‘–”‘Ž•ǡƒ†”‡†—…‡†Ž‡˜‡Ž•‘ˆ’Š‘•’Š‘”›Žƒ–‡†ȋ‹‰—”‡ͶȌǤ‹…‡‹–‹•–Š‡‘”‡–‹…ƒŽŽ›
’‘••‹„Ž‡–Šƒ––Š‡”‡†—…‡†Ž‡˜‡Ž•‘ˆ’ͳ͹ʹǦ‹–Š‡—–ƒ–‰Žƒ†•‹‰Š–„‡†—‡–‘ƒ
Ž‘™Ȁ”ƒ–‹‘ǡ™‡‡ƒ•—”‡†ǡǡƒ†Ž‡˜‡Ž•‹‡š–”ƒ…–•ˆ”‘™‹Ž†–›’‡
ƒ†—–ƒ–‰Žƒ†•Ǥƒ†Ž‡˜‡Ž•‹—–ƒ–‰Žƒ†•™‡”‡…‘’ƒ”ƒ„Ž‡–‘™‹Ž†–›’‡
…‘–”‘Ž•ǡ ™Š‹Ž‡ Ž‡˜‡Ž• ™‡”‡ •Ž‹‰Š–Ž› Š‹‰Š‡” ‘ –Š‡ —–ƒ– ‰Žƒ†•Ǥ Š‡•‡ †ƒ–ƒ
‹†‹…ƒ–‡ –Šƒ– ͵ ‹Š‹„‹–• †‡•’‹–‡ Ȁ ƒ† Ȁ ”ƒ–‹‘• –Šƒ– ™‘—Ž†
•–‹—Žƒ–‡ƒ…–‹˜ƒ–‹‘ǡƒ•‹™‹Ž†–›’‡‰Žƒ†•ȋ‹‰—”‡ͶȌǤ
‘ –‡•– –Š‡ Š›’‘–Š‡•‹• –Šƒ– ͵ ƒ…–‹˜ƒ–‹‘ ”‡’”‡••‡• …›…Ž‡Ȁ ‰‡‡
‡š’”‡••‹‘–Š”‘—‰Š–Š‡‹Š‹„‹–‹‘‘ˆǡ™‡–”‡ƒ–‡†™‹Ž†–›’‡ƒ†Pten–Š›”ǦȀǦ‹…‡™‹–Š
–Š‡ƒ…–‹˜ƒ–‘”ˆ‘”ˆ‘—”™‡‡•ǡƒ†–Š‡‹•‘Žƒ–‡†–Š‡‹”–Š›”‘‹†‰Žƒ†•Ǥ
–”‡ƒ–‡–‹…”‡ƒ•‡†’ͳ͹ʹǦŽ‡˜‡Ž•‘–‘Ž›‹™‹Ž†–›’‡‹…‡ǡ„—–ƒŽ•‘ȋƒ†–‘–Š‡
•ƒ‡ Ž‡˜‡Ž•Ȍ ‹ Pten–Š›”ǦȀǦ ‹…‡ǡ †‡•’‹–‡ –Š‡ ’‡”•‹•–‡– ƒ…–‹˜ƒ–‹‘ ƒ† ‹Š‹„‹–‘”›
’Š‘•’Š‘”›Žƒ–‹‘ ‘ Ͷͺͷ ȋ‹‰—”‡ ͷȌǤ —”–Š‡”‘”‡ǡ Ǧƒ…–‹˜ƒ–‡† ”‡•–‘”‡†’Š‘•’Š‘”›Žƒ–‹‘‘‡”͹ͻȋ‹‰—”‡ͷȌǤ
‡‡š–‡ƒ•—”‡†–Š‡‡š’”‡••‹‘‘ˆERRα ERRγ, PGC-1ა†PGC-1⋐–Š‡–Š›”‘‹†•
‘ˆ …‘–”‘Žǡ —–ƒ–ǡ ƒ† Ǧ–”‡ƒ–‡† —–ƒ– ‹…‡Ǥ –”‹‹‰Ž›ǡ –”‡ƒ–‡–
‹…”‡ƒ•‡† –Š‡  Ž‡˜‡Ž• ‘ˆ ƒŽŽ ˆ‘—” –”ƒ•…”‹’–‹‘ ˆƒ…–‘”• ƒ† …‘Ǧˆƒ…–‘”• –‘ ‡ƒ”Ǧ™‹Ž†
ͳ͸
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
–›’‡ Ž‡˜‡Ž• ȋ‹‰—”‡ ͷȌǤ ‘ ˜‡”‹ˆ› –Šƒ– –Š‡ ‹…”‡ƒ•‡ ‹ –”ƒ•…”‹’–‹‘ƒŽ ”‡‰—Žƒ–‘” Ž‡˜‡Ž•
–”ƒ•Žƒ–‡•‹ƒ”‡•–‘”ƒ–‹‘‘ˆ‘”ƒŽ…›…Ž‡Ȁ‰‡‡‡š’”‡••‹‘ǡ™‡‡ƒ•—”‡†
„› “ ƒ† ‡•–‡” „Ž‘––‹‰ –Š‡ ‡š’”‡••‹‘ Ž‡˜‡Ž• ‘ˆ •‡Ž‡…–‡† …›…Ž‡ ‰‡‡•ǡ ƒ†
ˆ‘—†–Šƒ––”‡ƒ–‡–Šƒ†…‘’Ž‡–‡Ž›”‡•…—‡†–Š‡͵Ǧ†‡’‡†‡–”‡’”‡••‹‘‘ˆ
Aco2ǡ Idh3aǡ ƒ† Pdhb ȋ‹‰—”‡ ͷǡȌǤ ‘™‡˜‡”ǡ ™Š‡ ™‡ –‡•–‡† –Š‡ ‡š’”‡••‹‘ ‘ˆ –Š‡
‹–‘…Š‘†”‹ƒǦ‡…‘†‡†‰‡‡•ǡ™‡…‘—Ž†‘Ž›†‡–‡…–ƒ’ƒ”–‹ƒŽ”‡•…—‡—’‘
–”‡ƒ–‡– ȋ‹‰—”‡ ͷȌǤ —”–Š‡”‘”‡ǡ –Š‡ ‡š–‡– ‘ˆ ‹–‘…Š‘†”‹ƒŽ †ƒƒ‰‡ ‘„•‡”˜‡† ‹
Pten–Š›”ǦȀǦ–Š›”‘…›–‡•™ƒ•‘Ž›‹Ž†Ž›”‡†—…‡†„›–”‡ƒ–‡–ȋ‹‰—”‡ͷȌǤ‡•’‹–‡
–Š‡’‡”•‹•–‡…‡‘ˆ•‘‡‹–‘…Š‘†”‹ƒŽ†ƒƒ‰‡ǡ‡ˆ‘”…‡†ƒ…–‹˜ƒ–‹‘”‡˜‡”•‡†–Š‡
‰Ž›…‘Ž›–‹…•™‹–…Š‹†—…‡†„›…‘•–‹–—–‹˜‡͵ƒ…–‹˜ƒ–‹‘ǡƒ••Š‘™„›–Š‡”‡†—…–‹‘‘ˆ
Žƒ…–ƒ–‡…‘–‡––‘™‹Ž†–›’‡Ž‡˜‡Ž•‹Ǧ–”‡ƒ–‡†Pten–Š›”ǦȀǦ‹…‡ȋ‹‰—”‡ͷ
ȌǤ
Š‡•‡†ƒ–ƒ•–”‘‰Ž›•—‰‰‡•––Šƒ–͵ȀǦ‡†‹ƒ–‡†‹Š‹„‹–‹‘‹•”‡•’‘•‹„Ž‡ˆ‘”
–Š‡”‡’”‡••‹‘‘ˆ‘š‹†ƒ–‹˜‡‡–ƒ„‘Ž‹•‰‡‡‡š’”‡••‹‘‘„•‡”˜‡†‹Pten–Š›”ǦȀǦ‹…‡Ǥ
AMPK inhibition actively contributes to thyroid hyperplasia.
‘–‡•–™Š‡–Š‡”–Š‡‰Ž›…‘Ž›–‹…•™‹–…Š‘„•‡”˜‡†‹Pten–Š›”ǦȀǦ‹…‡’Žƒ›•ƒƒ…–‹˜‡”‘Ž‡‹
–Š‡ Š›’‡”’”‘Ž‹ˆ‡”ƒ–‹˜‡ ƒ† ’”‘Ǧ–—‘”‹‰‡‹… ’Š‡‘–›’‡ ‘ˆ —–ƒ– –Š›”‘…›–‡•ǡ ™‡
…‘’ƒ”‡†–Š‡™‡‹‰Š–ƒ†’”‘Ž‹ˆ‡”ƒ–‹˜‡‹†‡š‘ˆ™‹Ž†–›’‡ǡPten–Š›”ǦȀǦǡƒ†Ǧ–”‡ƒ–‡†
—–ƒ– –Š›”‘‹† ‰Žƒ†•Ǥ —”‹‰ –Š‡ ˆ‘—” ™‡‡• ‘ˆ ‡ˆ‘”…‡† ”‡ƒ…–‹˜ƒ–‹‘ǡ —–ƒ–
‰Žƒ†•‰”‡™ƒ–ƒ—…Š•Ž‘™‡””ƒ–‡…‘’ƒ”‡†–‘—–”‡ƒ–‡†—–ƒ–•ǡ•‘–Šƒ––Š‡™‡‹‰Š–‘ˆ
–Š‡—–ƒ––Š›”‘‹†•ƒ––Š‡‡†‘ˆ–Š‡–”‡ƒ–‡–™ƒ••‹‰‹ˆ‹…ƒ–Ž›”‡†—…‡†…‘’ƒ”‡†–‘
—–”‡ƒ–‡† Pten–Š›”ǦȀǦ ‹…‡ ȋǦ͵ͶΨǡ ‹‰—”‡ ͷȌǤ Š‹• •‹œ‡ ”‡†—…–‹‘ ™ƒ• ƒ••‘…‹ƒ–‡† ™‹–Š
†‡…”‡ƒ•‡† ’”‘Ž‹ˆ‡”ƒ–‹‘ǣ –Š‡ –Š›”‘…›–‡ ’”‘Ž‹ˆ‡”ƒ–‹‘ ‹†‡š ‹ Ǧ–”‡ƒ–‡† ‹…‡ǡ
†‡–‡”‹‡† „› ”† ‹…‘”’‘”ƒ–‹‘ǡ ™ƒ• †”ƒ•–‹…ƒŽŽ› ”‡†—…‡†ǡ …‘’ƒ”‡† –‘ –Šƒ– ‘ˆ
ͳ͹
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
—–”‡ƒ–‡† —–ƒ– ‹…‡ ȋǦͷ͵Ψǡ ‹‰—”‡ ͷȌǤ Š—•ǡ ‹– ‹• ’‘••‹„Ž‡ –Šƒ– –Šƒ– –Š‡ ‰Ž›…‘Ž›–‹…
•™‹–…Š …ƒ—•‡† „› ͵Ǧ‡†‹ƒ–‡† ‹Š‹„‹–‹‘ ƒ…–‹˜‡Ž› …‘–”‹„—–‡• –‘ –Š‡
†‡˜‡Ž‘’‡–‘ˆ–Š›”‘‹†Š›’‡”’Žƒ•‹ƒ‹Pten–Š›”ǦȀǦ‹…‡Ǥ
The PI3K/AMPK mediated repression of oxidative metabolism is maintained in human
thyroid cancer cells and involves AMPK-mediated phosphorylation of PGC-1α.
ƒ˜‹‰‡•–ƒ„Ž‹•Š‡†‹ƒ”‡Ž‡˜ƒ–in vivo•›•–‡–Šƒ–•—•–ƒ‹‡†͵ƒ…–‹˜ƒ–‹‘”‡†‹”‡…–•
‡‡”‰› ‡–ƒ„‘Ž‹• –‘™ƒ”†• ‰Ž›…‘Ž›•‹• „› ”‡’”‡••‹‰ –Š‡ ‡š’”‡••‹‘ ‘ˆ …›…Ž‡ ƒ†
‰‡‡• ‹ ‘—•‡ –Š›”‘…›–‡•ǡ ™‡ •‘—‰Š– –‘ ‡š–‡† –Š‡•‡ •–—†‹‡• –‘ –Š‡ Š—ƒ
•‡––‹‰Ǥ ‡ Šƒ˜‡ ‹‹–‹ƒŽŽ› —•‡† –Š‡ PTENǦȀǦ ˆ‘ŽŽ‹…—Žƒ” –Š›”‘‹† …ƒ…‡” …‡ŽŽ Ž‹‡ǡ Ǧͳ͵͵ǣ
—–”‡ƒ–‡†ǡ ‘”ƒŽŽ› ‰”‘™‹‰ …‡ŽŽ• †‹•’Žƒ›‡† ƒŽ‘•– —†‡–‡…–ƒ„Ž‡ ’ͳ͹ʹǦǡ ƒ†
•–”‘‰ ’ͶͺͷǦǤ  ‡‡’‹‰ ™‹–Š –Š‡ ‘—•‡ †ƒ–ƒǡ –”‡ƒ–‡– ‹…”‡ƒ•‡†
’ͳ͹ʹǦǡƒ•™‡ŽŽƒ•’͹ͻǦȋƒ†‹”‡…––ƒ”‰‡–Ȍ‹ƒ†‘•‡Ǧ†‡’‡†‡–ƒ‡”
ȋ‹‰—”‡͸ȌǤ—”–Š‡”‘”‡ǡ‡ˆ‘”…‡†”‡ƒ…–‹˜ƒ–‹‘‘ˆŽ‡†–‘•‹‰‹ˆ‹…ƒ–Ž›‹…”‡ƒ•‡†
‡š’”‡••‹‘‘ˆ–Š‡…›…Ž‡‰‡‡•ACO2ǡIDH3Aǡƒ†PDHBȋ‹‰—”‡͸ȌǤ
• ‡–‹‘‡† ƒ„‘˜‡ǡ ‹‰Š– …‘–”‘Ž –Š‡ ‡š’”‡••‹‘ ƒ† ˆ—…–‹‘ ‘ˆ ‡–ƒ„‘Ž‹…
–”ƒ•…”‹’–‹‘ ˆƒ…–‘”• ƒ† ‡œ›‡• „› ‘†—Žƒ–‹‰ Ǧͳα ƒ…–‹˜‹–› ˜‹ƒ ’Š‘•’Š‘”›Žƒ–‹‘
‘ Ǧͳα ͳ͹͹ ƒ† ͷ͵ͺ ȋʹͷȌǤ ‘ –‡•– ™Š‡–Š‡” –Š‹• ’ƒ–Š™ƒ›ǡ †‹•…‘˜‡”‡† ‹ •‡Ž‡–ƒŽ
—•…Ž‡ …‡ŽŽ•ǡ ‹• ƒŽ•‘ ƒ…–‹˜‡ ‹ –Š›”‘…›–‡•ǡ ™‡ –”ƒ•ˆ‡…–‡† Ǧͳ͵͵ …‡ŽŽ• ™‹–Š ƒ
‡š’”‡••‹‘ ˜‡…–‘” ‡…‘†‹‰ ™‹Ž† –›’‡ ‘” ’Š‘•’Š‘”›Žƒ–‹‘ †‡ˆ‡…–‹˜‡ Ǧͳα ȋ
ǦͳαǦ
Ȍǡ –”‡ƒ–‡† –Š‡ …‡ŽŽ• ™‹–Š ˜‡Š‹…Ž‡ ‘” –‘ ”‡˜‡”•‡ –Š‡ ‹Š‹„‹–‹‘ ‘ˆ ǡ ƒ†
‡š–”ƒ…–‡†  –‘ ‡ƒ•—”‡ –Š‡ ‡š’”‡••‹‘ ‘ˆ ƒ ‰”‘—’ ‘ˆ …›…Ž‡ ‰‡‡•Ǥ ”‡ƒ…–‹˜ƒ–‹‘ ‹…”‡ƒ•‡† –Š‡ ‡š’”‡••‹‘ ‘ˆ ACO2ǡ IDH3Aǡ ƒ† PDHB ‹ …‡ŽŽ• –”ƒ•ˆ‡…–‡†
ͳͺ
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
™‹–Š™‹Ž†–›’‡
Ǧͳα„—–‘–ǡ‘”–‘ƒ—…ŠŽ‘™‡”‡š–‡–ǡ‹–Š‘•‡–”ƒ•ˆ‡…–‡†™‹–Š–Š‡
ǦͳᏗ–ƒ– –Šƒ– …ƒ‘– „‡ ’Š‘•’Š‘”›Žƒ–‡† ȋ‹‰—”‡ ͸ȌǤ Š—•ǡ Ǧͳα ƒ…–• ƒ• ƒ
…‘†—‹–ˆ‘”–‘…‘–”‘Ž–Š‡‡š’”‡••‹‘‘ˆ‡–ƒ„‘Ž‹…‰‡‡•Ǥ
 ‘”†‡” –‘ –‡•– ™Š‡–Š‡” ”‡’”‡••‹‘ ‹ Š—ƒ …‡ŽŽ• ‹• †‹”‡…–Ž› ƒ••‘…‹ƒ–‡† ™‹–Š
͵ ƒ…–‹˜ƒ–‹‘ǡ ™‡ –”‡ƒ–‡† Ǧͳ͵͵ …‡ŽŽ• ™‹–Š –Š‡ ’ƒǦ͵ ‹Š‹„‹–‘” ͳʹͲǤ
—”’”‹•‹‰Ž›ǡ͵‹Š‹„‹–‹‘ƒ–ƒ…‘…‡–”ƒ–‹‘–Šƒ–Šƒ•‘‘ˆˆǦ–ƒ”‰‡–‡ˆˆ‡…–•ȋͳρȌ
™ƒ•‘–•—ˆˆ‹…‹‡––‘ƒ„‘Ž‹•ŠͶͺͷ’Š‘•’Š‘”›Žƒ–‹‘ǡ‘”–‘‹†—…‡ͳ͹ʹ’Š‘•’Š‘”›Žƒ–‹‘
ȋ‹‰—”‡͸ȌǤ‘™‡˜‡”ǡŠ‹‰Š‡”ͳʹͲ…‘…‡–”ƒ–‹‘•…‘—Ž†ƒ…Š‹‡˜‡–Š‡•‡‡ˆˆ‡…–•ȋ†ƒ–ƒ
‘– •Š‘™Ȍ •—‰‰‡•–‹‰ –Šƒ– ‘–Š‡” ”‡Žƒ–‡† ‹ƒ•‡• ‹‰Š– …‘–”‹„—–‡ –‘ Ͷͺͷ
’Š‘•’Š‘”›Žƒ–‹‘ƒ†‹Š‹„‹–‹‘Ǥ
Šƒ•„‡‡•Š‘™–‘’Š‘•’Š‘”›Žƒ–‡ͶͺͷǦ‹–Š‡ ‹•—Ž‹Ǧ•‡…”‡–‹‰…‡ŽŽŽ‹‡ǡǦ
ͳ ȋʹͻȌǤ —”–Š‡”‘”‡ǡ ‹ ‘—•‡ ƒ†‹’‘…›–‡•ǡ …ƒ ’Š‘•’Š‘”›Žƒ–‡ ‘ ͳ͹͵ǡ
‹Š‹„‹–‹‰ͳ͹ʹ’Š‘•’Š‘”›Žƒ–‹‘ǡŽ‹‡Ž›–Š”‘—‰Š•–‡”‹…Š‹†”ƒ…‡ȋ͵ͲȌǤŠ—•ǡ™‡–‡•–‡†
™Š‡–Š‡” ‹Š‹„‹–‹‘ …‘—Ž† •›‡”‰‹œ‡ ™‹–Š ͵ ‹Š‹„‹–‹‘ –‘ ”‡•–‘”‡ ƒ…–‹˜‹–›
ƒ† ”‡•’‹”ƒ–‘”› ‰‡‡ ‡š’”‡••‹‘Ǥ –”‹‹‰Ž›ǡ ™Š‹Ž‡ Ǧͳ͵͵ –”‡ƒ–‡– ™‹–Š –Š‡ ‹Š‹„‹–‘”ͺͻƒŽ‘‡†‹†‘–”‡•—Ž–‹ƒŽ–‡”ƒ–‹‘•‘ˆ–Š‡’Š‘•’Š‘”›Žƒ–‹‘•–ƒ–—•‘ˆǡ
•‹—Ž–ƒ‡‘—• ‹Š‹„‹–‹‘ ‘ˆ ͵ ƒ† ƒ„‘Ž‹•Š‡† ’ͶͺͷǦ ƒ† †”ƒ•–‹…ƒŽŽ›
‹…”‡ƒ•‡† ’ͳ͹ʹǦ ȋ‹‰—”‡ ͸ȌǤ ……‘”†‹‰Ž›ǡ ™Š‹Ž‡ •‹‰Ž‡Ǧ‹Š‹„‹–‘” –”‡ƒ–‡– ‘ˆ
Ǧͳ͵͵ ˆƒ‹Ž‡† –‘ ‹…”‡ƒ•‡ –Š‡ ‡š’”‡••‹‘ ‘ˆ …›…Ž‡ ‰‡‡•ǡ ‘” Ǧͳα ƒ† Ǧͳβǡ
…‘„‹‡† –”‡ƒ–‡– •‹‰‹ˆ‹…ƒ–Ž› ‹…”‡ƒ•‡† –Š‡‹”  Ž‡˜‡Ž• ȋ‹‰—”‡ ͸ȌǤ ‘„‹‡†
‹Š‹„‹–‹‘ ‘ˆ ͵ ƒ† ™ƒ• ƒŽ•‘ ‘”‡ ‡ˆˆ‡…–‹˜‡ –Šƒ ‡‹–Š‡” ‹ƒ•‡ ‹Š‹„‹–‹‘ ‹
‹…”‡ƒ•‹‰–Š‡‡š’”‡••‹‘‘ˆƒŽ—…‹ˆ‡”ƒ•‡”‡’‘”–‡”†”‹˜‡„›–Š‡’”‘‘–‡””‡‰‹‘‘ˆPGC-
ͳͻ
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
1α,ˆ—”–Š‡”•—’’‘”–‹‰–Š‡”‘Ž‡‘ˆ‹–Š‡…‘–”‘Ž‘ˆ
Ǧͳαƒ…–‹˜‹–›ƒ†‡š’”‡••‹‘
ȋ‹‰—”‡͸ȌǤ
Š‡•‡†ƒ–ƒ•—‰‰‡•––Šƒ––Š‡͵Ȁƒš‹•…‘–”‘Ž•ƒ…–‹˜‹–›ƒŽ•‘‹PTENǦȀǦŠ—ƒ
–Š›”‘‹† …ƒ…‡” …‡ŽŽ•Ǣ Š‘™‡˜‡”ǡ ‹ –Š‡•‡ …‡ŽŽ•ǡ …‘–”‹„—–‡• –‘ –Š‡ ‡–ƒ„‘Ž‹…
”‡’”‘‰”ƒ‹‰–Š”‘—‰Šƒ‘˜‡”Žƒ’’‹‰’ƒ–Š™ƒ›Ǥ
‘–‡•–™Š‡–Š‡”–Š‹••‹‰ƒŽ‹‰…ƒ•…ƒ†‡‹•ƒŽ•‘ƒ…–‹˜‡‹Š—ƒƒƒ’Žƒ•–‹…–Š›”‘‹†…ƒ…‡”ǡ
™‡ —•‡† –™‘ …‡ŽŽ Ž‹‡• ”‡’”‡•‡–ƒ–‹˜‡ ‘ˆ –Š‡ ‘•– …‘‘ †”‹˜‡” ’ƒ–Š™ƒ›• ‹ –Š‹•
ƒ‰‰”‡••‹˜‡ƒ†‰‡‡–‹…ƒŽŽ›…‘’Ž‡š–—‘”–›’‡ǣͳ͸ǡŠƒ”„‘”‹‰ƒƒ…–‹˜ƒ–‹‰PIK3CA
—–ƒ–‹‘ȋͷͶͷȌǡƒ†ͺͷͲͷ…ǡŠƒ”„‘”‹‰–Š‡BRAF‘…‘‰‡‹…ƒŽŽ‡Ž‡͸ͲͲǤ
‘–Š …‡ŽŽ Ž‹‡• †‹•’Žƒ›‡† Ž‘™ –‘ —†‡–‡…–ƒ„Ž‡ ’ͳ͹ʹǦǡ ƒ• ™‡ŽŽ ƒ• •–”‘‰ ’ͶͺͷǦ
ǡ •—‰‰‡•–‹‰ –Šƒ– ‹Š‹„‹–‹‘ ‹• ƒ …‘‘ –Š‡‡ ‹ –Š›”‘‹† …ƒ…‡” ȋ‹‰—”‡
͸
ȌǤ–”‡ƒ–‡–”‡•–‘”‡†ͳ͹ʹ’Š‘•’Š‘”›Žƒ–‹‘ǡ‘”‡‡ˆˆ‡…–‹˜‡Ž›‹–Š‡…‡ŽŽŽ‹‡
Šƒ”„‘”‹‰–Š‡͵‘…‘‰‡‹…—–ƒ–‹‘ȋ‹‰—”‡͸
ȌǤ
Š‡™‡–”‡ƒ–‡†–Š‡•‡…‡ŽŽ•™‹–Š͵ƒ†‹Š‹„‹–‘”•ǡƒŽ‘‡ƒ†‹…‘„‹ƒ–‹‘ǡ™‡
ˆ‘—† –Šƒ– •‹—Ž–ƒ‡‘—• ͵ ƒ† ‹Š‹„‹–‹‘ ‡ˆˆ‡…–‹˜‡Ž› ”‡Ǧƒ…–‹˜ƒ–‡† ƒ†
‹…”‡ƒ•‡†–Š‡‡š’”‡••‹‘‘ˆ‡–ƒ„‘Ž‹…”‡‰—Žƒ–‘”••—…Šƒ•ERRバ†PGC-1⋐…‡ŽŽ•™‹–Š
…‘•–‹–—–‹˜‡Ž› ƒ…–‹˜‡ ͵Ǣ Š‘™‡˜‡”ǡ –Š‡› †‹† ‘– ƒˆˆ‡…– ƒ…–‹˜ƒ–‹‘ ‘” ERRγ ƒ†
PGC-1β‡š’”‡••‹‘‹–Š‡BRAF͸ͲͲ…‡ŽŽŽ‹‡ȋ‹‰—”‡͸ǡȌǤ‹‹Žƒ”–‘–Š‡in vivo‘†‡Žǡ
”‡ƒ…–‹˜ƒ–‹‘”‡†—…‡†’”‘Ž‹ˆ‡”ƒ–‹‘‘ˆƒ’ƒ‡Ž‘ˆ‘—•‡ƒ†Š—ƒ–Š›”‘‹†…ƒ…‡”
…‡ŽŽŽ‹‡•…ƒ””›‹‰͵ƒ…–‹˜ƒ–‹‰—–ƒ–‹‘•ȋ‹‰—”‡͸ȌǤ
Š—•ǡ ‹ ™‡ŽŽǦ†‹ˆˆ‡”‡–‹ƒ–‡† ƒ† —†‹ˆˆ‡”‡–‹ƒ–‡† Š—ƒ –Š›”‘‹† …ƒ…‡” …‡ŽŽ•ǡ
…‘•–‹–—–‹˜‡ƒ…–‹˜ƒ–‹‘‘ˆ͵ǡ‹…‘‘’‡”ƒ–‹‘™‹–ŠǡƒŽ–‡”•–Š‡‡š’”‡••‹‘‘ˆ‰‡‡•
‹˜‘Ž˜‡†‹”‡•’‹”ƒ–‘”›‡–ƒ„‘Ž‹•„›‹Š‹„‹–‹‰Ǥ
ʹͲ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
TCA cycle gene repression in neoplastic lesions is also observed in other human tissues.
‘ ‡š–‡† ‘—” ˆ‹†‹‰• –‘ Š—ƒ –—‘”• ‘–Š‡” –Šƒ –Š‘•‡ ƒ”‹•‹‰ ‹ –Š‡ –Š›”‘‹†ǡ ™‡
‹–‡””‘‰ƒ–‡† –Š‡ …‘‹‡ †ƒ–ƒ„ƒ•‡ ˆ‘” †ƒ–ƒ•‡–• •Š‘™‹‰ •‹‰‹ˆ‹…ƒ– •‹—Ž–ƒ‡‘—•
†‘™Ǧ”‡‰—Žƒ–‹‘ ‘ˆ „‘–Š PTEN ƒ† …›…Ž‡ ‰‡‡•Ǥ Ž–Š‘—‰Š –Š‹• ƒ’’”‘ƒ…Š †‘‡• ‘–
ƒ……‘—– ˆ‘” ‘–Š‡”ǡ ‘”‡ …‘‘ǡ ͵Ǧƒ…–‹˜ƒ–‹‰ ‡…Šƒ‹••ǡ ‹– ”‡’”‡•‡–• –Š‡ ‘Ž›
†‹”‡…– ™ƒ› –‘ ‹†‡–‹ˆ›ǡ ™‹–Š‹ Žƒ”‰‡ ‡š’”‡••‹‘ †ƒ–ƒ•‡–•ǡ –—‘”• ™‹–Š ƒ…–‹˜ƒ–‡† ͵
•‹‰ƒŽ‹‰ǡ‹–Š‡ƒ„•‡…‡‘ˆ˜ƒŽ‹†ƒ–‡†Dz͵ƒ…–‹˜ƒ–‹‘dz•‹‰ƒ–—”‡•Ǥ
‘•‹†‡”‹‰–Š‡‹Š‡”‡–Ž‘™’‘™‡”‘ˆ‘—”•‡ƒ”…Šǡ‹–‹•”‡ƒ”ƒ„Ž‡–Šƒ–™‡ˆ‘—†ƒ˜‡”›
•–”‘‰ƒ••‘…‹ƒ–‹‘„‡–™‡‡PTENŽ‘••ƒ†…›…Ž‡”‡’”‡••‹‘‹–™‘†ƒ–ƒ•‡–•ǡ†‡”‹˜‡†
ˆ”‘†‡†‹ˆˆ‡”‡–‹ƒ–‡†Ž‹’‘•ƒ”…‘ƒƒ†ˆ”‘‡ƒ”Ž›•–ƒ‰‡…‘Ž‘…ƒ…‡”ȋ—’’ŽǤ‹‰—”‡ͳȌǤ
Š—•ǡ ‘–Š‡” Š—ƒ –—‘” –›’‡• †‹•’Žƒ› ‰Ž‘„ƒŽ †‘™Ǧ”‡‰—Žƒ–‹‘ ‘ˆ …›…Ž‡ ‰‡‡• ‹
ƒ••‘…‹ƒ–‹‘ ™‹–Š PTEN Ž‘•• ƒ† ͵ ƒ…–‹˜ƒ–‹‘ǡ ™ƒ””ƒ–‹‰ ˆ—–—”‡ •–—†‹‡• –‘
‡š’‡”‹‡–ƒŽŽ›˜ƒŽ‹†ƒ–‡–Š‹•ƒ••‘…‹ƒ–‹‘Ǥ
ʹͳ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
DISCUSSION
Š‡‡š‹•–‡…‡‘ˆƒ–‹‰Š–Ž‹„‡–™‡‡͵•‹‰ƒŽ‹‰ƒ†”‡”‘—–‹‰‘ˆ‡‡”‰›‡–ƒ„‘Ž‹•
‹• ™‡ŽŽ ‡•–ƒ„Ž‹•Š‡† ‹ …ƒ…‡” …‡ŽŽ•ǡ ™Š‡”‡ ƒ…–‹˜ƒ–‹‘ ‘ˆ ͵ …‘–”‹„—–‡• –‘ –Š‡ •™‹–…Š
ˆ”‘ ‘š‹†ƒ–‹˜‡ –‘ ‰Ž›…‘Ž›–‹… ’ƒ–Š™ƒ›• –Š”‘—‰Š •‡˜‡”ƒŽ ‘ˆ ‹–• †‘™•–”‡ƒ ‡ˆˆ‡…–‘”•ǡ
‹…Ž—†‹‰ǡǡƒ†ͳαȋ͵ͳȌǤ–—”ǡ–Š‡•‡‡ˆˆ‡…–‘”•‹…”‡ƒ•‡–Š‡‡š’”‡••‹‘
ƒ†Ȁ‘” ƒ…–‹˜‹–› ‘ˆ ƒ ˜ƒ”‹‡–› ‘ˆ ‰Ž›…‘Ž›–‹… ‡œ›‡•ǡ •—…Š ƒ• ‡š‘‹ƒ•‡ ȋ͵ʹȌǡ
Š‘•’Š‘ˆ”—…–‘‹ƒ•‡ ʹ ȋ͵͵Ȍǡ –Š‡ ˆƒ‹Ž› ‘ˆ ‰Ž—…‘•‡ –”ƒ•’‘”–‡”• ȋ͵ͶȌǡ ƒ…–ƒ–‡
†‡Š›†”‘‰‡ƒ•‡• ȋ͵ͷǡ ͵͸Ȍǡ ƒ† –Š‡ ʹ ‹•‘ˆ‘” ‘ˆ ›”—˜ƒ–‡ ‹ƒ•‡ ȋʹ͵ȌǤ Š—•ǡ ‹ ˆ—ŽŽ›
–”ƒ•ˆ‘”‡†…‡ŽŽ•ǡ–Š‡ƒ”„—”‰‡ˆˆ‡…–‹•ƒ…Š‹‡˜‡†–Š”‘—‰Šƒ‹…”‡ƒ•‡‹‰Ž›…‘Ž›–‹…ˆŽ—šǡ
™Š‹Ž‡‹–‘…Š‘†”‹ƒŽ‡–ƒ„‘Ž‹•‹•—ƒˆˆ‡…–‡†Ǥ
‹––Ž‡ ‹• ‘™ ‘ˆ –Š‡ ‡…Šƒ‹•ȋ•Ȍ ”‡‰—Žƒ–‹‰ ‡‡”‰›‡–ƒ„‘Ž‹• ‹ ‘”ƒŽ ƒ† ’”‡Ǧ
‡‘’Žƒ•–‹… ȋ‹Ǥ‡Ǥ …ƒ””›‹‰ •‹‰Ž‡ —–ƒ–‹‘• ƒ† ‘– ›‡– –”ƒ•ˆ‘”‡†Ȍ …‡ŽŽ•Ǥ Š‡”‡ ‹• ƒ
…‘•‡•—•–Šƒ–‹‘”ƒŽ…‡ŽŽ•–Š‡‰Ž›…‘Ž›–‹…•™‹–…Š‹•Ž‹‡†–‘Š‹‰Š’”‘Ž‹ˆ‡”ƒ–‹‘”ƒ–‡•ǡ
ƒ••Š‘™ˆ‘”‡„”›‘‹…–‹••—‡•ƒ†ƒ…–‹˜ƒ–‡†Ž›’Š‘…›–‡•ȋ͵͹ȌǤŠ‹Ž‡ƒ‡Ž‡‰ƒ–•–—†›
Šƒ• ”‡…‡–Ž› ‡•–ƒ„Ž‹•Š‡† –Šƒ– ƒ…–‹˜ƒ–‡† Ž›’Š‘…›–‡• ”‡Ž› ƒ‹Ž› ‘ –‘
–”ƒ•…”‹’–‹‘ƒŽŽ›”‡’”‘‰”ƒ–Š‡‹”‡–ƒ„‘Ž‹•ȋ͵ͺȌǡ—…ŠŽ‡••‹•‘™™Š‡‹–…‘‡•
–‘‡’‹–Š‡Ž‹ƒŽ…‡ŽŽ•Ǥ
—”†ƒ–ƒǡ‰‡‡”ƒ–‡†‹ƒ”‡Ž‡˜ƒ–in vivo‘†‡Žǡ•—’’‘”––Š‡Š›’‘–Š‡•‹•–Šƒ–ǡ‹”‡•’‘•‡
–‘ ͵ ƒ…–‹˜ƒ–‹‘ǡ ‘Ǧ–”ƒ•ˆ‘”‡† –Š›”‘‹† ‡’‹–Š‡Ž‹ƒŽ …‡ŽŽ• ‹…”‡ƒ•‡ ‡–ƒ„‘Ž‹… ˆŽ—š
–Š”‘—‰Š‰Ž›…‘Ž›•‹•ǡƒ•‹†‹…ƒ–‡†„›–Š‡†”ƒƒ–‹…‹…”‡ƒ•‡‹Žƒ…–ƒ–‡’”‘†—…–‹‘‘„•‡”˜‡†
‹—–ƒ–…‡ŽŽ•ǢŠ‘™‡˜‡”–Š‹•‡–ƒ„‘Ž‹…•™‹–…Š‹•‹†‡’‡†‡–‘ˆ–Š›”‘…›–‡’”‘Ž‹ˆ‡”ƒ–‹‘Ǥ
‡–ƒ„‘Ž‹… ”‡’”‘‰”ƒ‹‰ ‘ˆ –Š›”‘‹† ‡’‹–Š‡Ž‹ƒŽ …‡ŽŽ• ‹• ƒ…Š‹‡˜‡† –Š”‘—‰Š ƒ ’”‡˜‹‘—•Ž›
—‘™ ‡…Šƒ‹•ǡ ‹˜‘Ž˜‹‰ –Š‡ …‘‘”†‹ƒ–‡† †‘™Ǧ”‡‰—Žƒ–‹‘ ‘ˆ –Š‡ ‡š’”‡••‹‘ ‘ˆ
ʹʹ
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
…›…Ž‡ ƒ† ‰‡‡•ǡ ƒ† Ž‡ƒ†‹‰ –‘ †›•ˆ—…–‹‘ƒŽ ‹–‘…Š‘†”‹ƒ ƒ† ”‡†—…‡†
ƒ„‹Ž‹–› –‘ ’‡”ˆ‘” ”‡•’‹”ƒ–‘”› ‡–ƒ„‘Ž‹•Ǥ Š‹Ž‡ ™‡ †‹† ‘– ‘„•‡”˜‡ ƒ› •‹‰‹ˆ‹…ƒ–
‹…”‡ƒ•‡‹–Š‡‡š’”‡••‹‘‘ˆ‰Ž›…‘Ž›–‹…‰‡‡•ȋ™‹–Š–Š‡‡š…‡’–‹‘‘ˆGlkȌ—’‘PtenŽ‘••ǡ
™‡ ˆ‘—† ƒ ”‡’”‘†—…‹„Ž‡ —’Ǧ”‡‰—Žƒ–‹‘ ‘ˆ –Š‡ Glut1 –”ƒ•’‘”–‡”Ǥ ……‘”†‹‰Ž›ǡ ƒ •–”‘‰
…‘‡…–‹‘ „‡–™‡‡ Ž‘•• ‘ˆ ‡š’”‡••‹‘ ƒ† ‹…”‡ƒ•‡† ‡š’”‡••‹‘ ‘ˆ ͳ Šƒ•
„‡‡”‡…‡–Ž›”‡’‘”–‡†‹–Š›”‘‹†–—‘”•†‹•…‘˜‡”‡††—”‹‰—”‡Žƒ–‡† ͳͺ
Ǧ•…ƒ•
ȋ͵ͻȌǤ
—” †ƒ–ƒ ƒ”‡ ‹ ’ƒ”–‹ƒŽ ƒ‰”‡‡‡– ™‹–Š –Š‘•‡ ”‡…‡–Ž› ”‡’‘”–‡† „› –Š‡ ƒ†‘Žˆ‹ ‰”‘—’
—•‹‰ ƒ ‘—•‡ ‘†‡Ž ™‹–Š ‡ˆ‘”…‡† ™Š‘Ž‡Ǧ„‘†› ‘˜‡”‡š’”‡••‹‘ ‘ˆ Pten ȋͶͲȌǤ •
ˆ”‘ –Š‡•‡ ‹…‡ †‹•’Žƒ› ƒ ‡–ƒ„‘Ž‹… •Š‹ˆ– –‘™ƒ”†• ǡ †‡…”‡ƒ•‡† Žƒ…–ƒ–‡
’”‘†—…–‹‘ǡ ƒ† ‹…”‡ƒ•‡† ‡š’”‡••‹‘ ‘ˆ ǦͳαǦ–ƒ”‰‡– ‰‡‡•Ǥ ‘™‡˜‡”ǡ ‹ •–”‹‹‰
…‘–”ƒ•–™‹–Š‘—”‘†‡Žǡ…‡ŽŽ•‘˜‡”‡š’”‡••‹‰PtenŠƒ˜‡‹…”‡ƒ•‡†‹–‘…Š‘†”‹ƒ—„‡”
ƒ†—’Ǧ”‡‰—Žƒ–‡PKM2–Š”‘—‰ŠǤŠ‹Ž‡–Š‡•‡†‹ˆˆ‡”‡…‡•‹‰Š–„‡Ž‹‡†–‘–‹••—‡Ǧ
•’‡…‹ˆ‹…‹–›‘ˆ •‘‡ ‘ˆ –Š‡ …‘–”‘ŽŽ‹‰ ’ƒ–Š™ƒ›•ǡ –Š‡› ‹‰Š– ƒŽ•‘ „‡ –Š‡ …‘•‡“—‡…‡ ‘ˆ
‘˜‡”‡š’”‡••‹‰ –‡ǡ ™Š‹…Š …‘’Ž‡–‡Ž› ƒ„‘Ž‹•Š‡• ͵ •‹‰ƒŽ‹‰Ǥ ‘–ƒ„Ž›ǡ –Š‡
‡š’”‡••‹‘ ‘ˆ ‰‡‡• ‹ Pten,Pdk1–Š›”ǦȀǦ ‹…‡ ȋ‹ ™Š‹…Š ͵ •‹‰ƒŽ‹‰ ‹•
†”ƒ•–‹…ƒŽŽ› ”‡’”‡••‡†ǡ Ž‹‡ ‹ –Š‡ Pten –”ƒ•‰‡‹… •–”ƒ‹Ȍ ™ƒ• Š‹‰Š‡” –Šƒ ‹ ™‹Ž† –›’‡
…‘–”‘Žǡ•—’’‘”–‹‰–Š‡Žƒ––‡”Š›’‘–Š‡•‹•Ǥ
”‘ ƒ ‡…Šƒ‹•–‹… •–ƒ†’‘‹–ǡ ‘—” †ƒ–ƒ •—’’‘”– ƒ ‘†‡Ž ‹ ™Š‹…Š ͵ ƒ…–‹˜ƒ–‹‘
‹‹–‹ƒ–‡•–Š‡”‡‘†‡Ž‹‰‘ˆ‡‡”‰›‡–ƒ„‘Ž‹•–Š”‘—‰Š–Š‡’Š‘•’Š‘”›Žƒ–‹‘‘ˆ‘
Ͷͺͷƒ†‹–•…‘•‡“—‡–‹ƒ…–‹˜ƒ–‹‘Ǥ‹•ƒ‹ƒ•‡‘™–‘’Žƒ›…”‹–‹…ƒŽ”‘Ž‡•‹
‰”‘™–Šǡ ‡–ƒ„‘Ž‹•ǡ ƒ—–‘’Šƒ‰›ǡ ƒ† …‡ŽŽ ’‘Žƒ”‹–› ȋͶͳǡ ͶʹȌǤ –• ƒ…–‹˜ƒ–‹‘ ”‡“—‹”‡•
’Š‘•’Š‘”›Žƒ–‹‘‘ͳ͹ʹ„›ͳȋͶͳȌǤ—…Š’Š‘•’Š‘”›Žƒ–‹‘…ƒ„‡‹Š‹„‹–‡†„›–Š‡
ʹ͵
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Ǧ ƒ† Ǧ‡†‹ƒ–‡† ’Š‘•’Š‘”›Žƒ–‹‘ ‘ˆ Ͷͺͷ ȋʹͻǡ Ͷ͵Ȍ ƒ† „› Ǧ‡†‹ƒ–‡†
’Š‘•’Š‘”›Žƒ–‹‘ ‘ˆ ͳ͹͵ ȋ͵ͲȌǤ ”‡…‡– ”‡’‘”– Šƒ• •Š‘™ –Šƒ– ƒŽ•‘ ͸ͳ …ƒ ‹Š‹„‹–
„› ’Š‘•’Š‘”›Žƒ–‹‰ Ͷͺͷ ȋͶͶȌǤ ‘™‡˜‡”ǡ –Š‡ ˆƒ…– –Šƒ– ͲͲͳ –”‡ƒ–‡– ‘ˆ
—–ƒ– ‹…‡ †‘‡• ‘– ”‡•…—‡ ƒ› ‘ˆ –Š‡ ‡–ƒ„‘Ž‹… ’Š‡‘–›’‡• †‡•’‹–‡ –Š‡ …‘’Ž‡–‡
ƒ„Žƒ–‹‘‘ˆ͸ƒ…–‹˜‹–›ƒ”‰—‡•ƒ‰ƒ‹•–ƒ”‘Ž‡ˆ‘”͸ͳ‹–Š‡’Š‘•’Š‘”›Žƒ–‹‘‘ˆǡ
ƒ–Ž‡ƒ•–‹–Š‡–Š›”‘‹†Ǥ‘”–Š‡•ƒ‡”‡ƒ•‘ǡƒŽ–Š‘—‰Š™ƒ•ˆ‘—†–‘„‡…‡–”ƒŽ–‘
–Š‡‰Ž›…‘Ž›–‹…•™‹–…Š‹Tsc2ǦȀǦ‘—•‡‹†‡›–—‘”•ƒ†‹PtenǦȀǦ•ȋʹ͵Ȍǡ‘—”in vivo
†ƒ–ƒ…Ž‡ƒ”Ž›•Š‘™–Šƒ–ƒ…–‹˜ƒ–‹‘‹•†‹•’‡•ƒ„Ž‡ˆ‘”–Š‡‡–ƒ„‘Ž‹…”‡‘†‡Ž‹‰‘ˆ
–Š‡’”‡Ǧ‡‘’Žƒ•–‹…–Š›”‘‹†Ǥ
Šƒ• „‡‡ ”‡…‡–Ž› •Š‘™ –‘ •—’’”‡•• –Š‡ ƒ”„—”‰ ‡ˆˆ‡…– ‹ ›…Ǧ†”‹˜‡
Ž›’Š‘ƒ•ǡ ƒ† ‹–• ‰‡‡–‹… ‹ƒ…–‹˜ƒ–‹‘ ‹†—…‡• ƒ ‰Ž›…‘Ž›–‹… •Š‹ˆ– ‹ ‘—•‡ ‡„”›‘‹…
ˆ‹„”‘„Žƒ•–• ȋȌ ˜‹ƒ ‘”‘š‹… •–ƒ„‹Ž‹œƒ–‹‘ ‘ˆ Ǧͳα ȋͶͷȌǤ ‘˜‡”•‡Ž›ǡ ™‡ †‘ ‘–
†‡–‡…– …Šƒ‰‡• ‹ Ǧͳα ’”‘–‡‹ Ž‡˜‡Ž• ‹ –Š‡ –Š›”‘‹†• ‘ˆ Pten–Š›”ǦȀǦ ‹…‡ǡ ‘” —’Ǧ
”‡‰—Žƒ–‹‘‘ˆLdha‘”AldoaǤ‘”‡‹’‘”–ƒ–Ž›ǡƒ…—–‡ƒ„Žƒ–‹‘†‹†‘–ƒˆˆ‡…–
‹„‘–ŠŽ›’Š‘ƒ•ƒ†•ǡ•—‰‰‡•–‹‰–Š‡ƒ„•‡…‡‘ˆƒ›†‡–”‹‡–ƒŽ‡ˆˆ‡…–•‘–Š‡
…›…Ž‡Ǥ Š‡•‡ †‹ˆˆ‡”‡…‡• ™‹–Š ‘—” ‘†‡Ž ‹‰Š– „‡ ”‡Žƒ–‡† –‘ –Š‡ …‘’Ž‡–‡ Ž‘•• ‘ˆ
˜•Ǥ ‹–• †‡…”‡ƒ•‡† ’Š‘•’Š‘”›Žƒ–‹‘ǡ ‘” ”‡ˆŽ‡…– –‹••—‡Ǧ•’‡…‹ˆ‹… ™‹”‹‰ ‘ˆ –Š‡
‡–ƒ„‘Ž‹…’ƒ–Š™ƒ›•Ǥ
•ƒ†
Ǧͳƒ”‡ƒŒ‘””‡‰—Žƒ–‘”•‘ˆ–Š‡‡š’”‡••‹‘‘ˆ…›…Ž‡Ȁ‰‡‡•ǡƒ†
–Š‡‹”Ž‘••‘”‘˜‡”‡š’”‡••‹‘Šƒ˜‡„‡‡•Š‘™–‘Ž‡ƒ†–‘”‡’”‡••‹‘‘”‹†—…–‹‘‘ˆ–Š‡•‡
‡–ƒ„‘Ž‹…–ƒ”‰‡–•ǡ”‡•’‡…–‹˜‡Ž›ȋʹͶǡͶ͸ǡͶ͹ȌǤ
ƒ…–‹˜ƒ–‹‘‘ˆ„›”‡•—Ž–•‹‹–•‹ƒ„‹Ž‹–›–‘–”ƒ•ƒ…–‹˜ƒ–‡
ǦͳαȋʹͷȌǡ™Š‹…Š
‹–—”Ž‡ƒ†•–‘Ž‘™‡”Ž‡˜‡Ž•‘ˆαȋʹ͸ȌǤ‡••…Ž‡ƒ”ǡ‹•–‡ƒ†ǡ‹•Š‘™…‘–”‘Ž•–Š‡
ʹͶ
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
‡š’”‡••‹‘‘ˆ
Ǧͳβǡ™Š‹…ŠŽƒ…•–Š‡–™‘”‡•‹†—‡•’Š‘•’Š‘”›Žƒ–‡†„›‹
Ǧͳαǡ
ƒ† –Šƒ– ‘ˆ γǤ Š‹Ž‡ ɀ †‘™Ǧ”‡‰—Žƒ–‹‘ ‹‰Š– „‡ †‹”‡…–Ž› ”‡•’‘•‹„Ž‡ ˆ‘” –Š‡
”‡†—…‡† Ž‡˜‡Ž• ‘ˆ Ǧͳβ ȋʹͶȌǡ ‹–• Ž‹ –‘ ͵ ƒ† ‹• •–‹ŽŽ —…Ž‡ƒ”Ǥ – Šƒ• „‡‡
”‡…‡–Ž› ”‡’‘”–‡† –Šƒ– ʹ —’Ǧ”‡‰—Žƒ–‡• ȋŽ‹‡Ž› –Š”‘—‰Š ͵Ȍ ‹Ǧ͵͹ͺȗ ‡š’”‡••‹‘
‹„”‡ƒ•–…ƒ…‡”…‡ŽŽ•ȋͶͺȌǤ–—”ǡ‹͵͹ͺȗ–ƒ”‰‡–•ERRǡŽ‡ƒ†‹‰–‘†‘™Ǧ”‡‰—Žƒ–‹‘‘ˆ
…›…Ž‡‰‡‡•ǡ”‡†—…‡†”‡•’‹”ƒ–‹‘”ƒ–‡ǡƒ†‹…”‡ƒ•‡†‰Ž›…‘Ž›•‹•Ǥ—–—”‡•–—†‹‡•™‹ŽŽ
‡˜ƒŽ—ƒ–‡ ™Š‡–Š‡” ͵ ƒ…–‹˜ƒ–‹‘ ‹ –Š›”‘‹† …‡ŽŽ• –ƒ”‰‡–• ERR –Š”‘—‰Š ‹Ǧ͵͹ͺȗǡ ƒ†
™Š‡–Š‡”’Žƒ›•ƒ”‘Ž‡‹–Š‹•’ƒ–Š™ƒ›Ǥ
Š‡ˆƒ…––Šƒ––”‡ƒ–‡–”‡•…—‡†‘Ž›’ƒ”–‹ƒŽŽ›–Š‡‡š’”‡••‹‘‘ˆ–Ǧ‡…‘†‡†
‰‡‡• ƒ† –Š‡ ƒŽ–‡”‡† ‹–‘…Š‘†”‹ƒ ‘”’Š‘Ž‘‰› ‹ Pten–Š›”ǦȀǦ ‹…‡ ‹‰Š– „‡ ƒ
…‘•‡“—‡…‡ ‘ˆ –Š‡ ‡š–‡•‹˜‡ †ƒƒ‰‡ •—•–ƒ‹‡† „› –Š‡•‡ ‘”‰ƒ‡ŽŽ‡•Ǥ Ž–‡”ƒ–‹˜‡Ž›ǡ
ƒ††‹–‹‘ƒŽ͵Ǧ†”‹˜‡’ƒ–Š™ƒ›•‹‰Š–…‘–”‹„—–‡–‘–Š‹•ƒ•’‡…–‘ˆ–Š‡’Š‡‘–›’‡Ǥ
ƒ††‹–‹‘ƒŽ •–‹ŽŽ ‘’‡ “—‡•–‹‘ ‹• ™Š› ƒˆˆ‡…–• ƒ…–‹˜‹–› ‹ –Š›”‘‹† …ƒ…‡” …‡ŽŽ
Ž‹‡•ǡ „—– †‘‡• ‘–ƒ’’‡ƒ” –‘†‘ •‘ in vivoǡ ‹ ‘—•‡ –Š›”‘…›–‡•Ǥ —”–Š‡” ™‘” ™‹ŽŽ „‡
‡‡†‡†–‘…Žƒ”‹ˆ›–Š‡•‡‹••—‡•Ǥ
‡ ‹’‘”–ƒ– ƒ•’‡…– ‘ˆ ‘—” ˆ‹†‹‰• ‹• –Šƒ– –Š‡ ‡–ƒ„‘Ž‹… ”‡‘†‡Ž‹‰ ƒ’’‡ƒ”• –‘
…‘–”‹„—–‡ †‹”‡…–Ž› –‘ –Š‡ Š›’‡”’”‘Ž‹ˆ‡”ƒ–‹˜‡ ’Š‡‘–›’‡ǡ •‹…‡ Ǧ‡†‹ƒ–‡†
”‡•–‘”ƒ–‹‘‘ˆ‘”ƒŽŽ‡˜‡Ž•‘ˆ…›…Ž‡‡œ›‡•ƒ†”‡†—…–‹‘‘ˆ‰Ž›…‘Ž›–‹…”ƒ–‡in vivo
†”ƒ•–‹…ƒŽŽ›”‡†—…‡•–Š›”‘…›–‡’”‘Ž‹ˆ‡”ƒ–‹‘Ǥ‘™‡˜‡”ǡ™‡…ƒ‘–…‘’Ž‡–‡Ž›‡š…Ž—†‡–Šƒ–
–Š‡ ”‡†—…‡† ’”‘Ž‹ˆ‡”ƒ–‹‘ ‹ Ǧ–”‡ƒ–‡† ‹…‡ ‹• ƒŽ•‘ –Š‡ …‘•‡“—‡…‡ ‘ˆ ƒ ‡‡”‰›Ǧ
†‡’”‹˜‡†•–ƒ–—•†—‡–‘’‡”•‹•–‡–‹–‘…Š‘†”‹ƒŽ†ƒƒ‰‡ƒ††›•ˆ—…–‹‘‹…‘†‹–‹‘•
ȋ‹Ǥ‡Ǥ–”‡ƒ–‡–Ȍ –Šƒ–†‡…”‡ƒ•‡–Š‡‡–ƒ„‘Ž‹…ˆŽ—š–Š”‘—‰Š‰Ž›…‘Ž›•‹•Ǥ
ʹͷ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
–‹•–‡’–‹‰–‘’”‘’‘•‡–Šƒ––Š‡‘Ǧ…ƒ‘‹…ƒŽ‰Ž›…‘Ž›–‹…•™‹–…Š–Šƒ––ƒ‡•’Žƒ…‡‹’”‡Ǧ
‡‘’Žƒ•–‹… PTENǦȀǦ …‡ŽŽ• ‹‰Š– …‘–”‹„—–‡ –‘ –Š‡ ‡‘’Žƒ•–‹… –”ƒ•ˆ‘”ƒ–‹‘ ‘ˆ –Š›”‘‹†ǡ
„”‡ƒ•–ǡ ƒ† …‘Ž‘ ‡’‹–Š‡Ž‹ƒŽ …‡ŽŽ• ‹ ‘™†‡ ‹•‡ƒ•‡ ’ƒ–‹‡–•ǡ …ƒ””›‹‰ ‰‡”Ž‹‡ PTEN
—–ƒ–‹‘•Ǥ‹‹Žƒ”–‘ SDHB‘”SDHD—–ƒ–‹‘…ƒ””‹‡”•ǡPTEN —–ƒ– ‘™†‡‹•‡ƒ•‡
’ƒ–‹‡–•Šƒ˜‡”‡…‡–Ž›„‡‡ˆ‘—†–‘Šƒ˜‡‡Ž‡˜ƒ–‡†’Žƒ•ƒ•—……‹ƒ–‡ȋͶͻȌǤŠ‡†‘™Ǧ
”‡‰—Žƒ–‹‘ ‘ˆ —……‹ƒ–‡ †‡Š›†”‘‰‡ƒ•‡• ‡š’”‡••‹‘ ‘„•‡”˜‡† ‹ –Š‡ –Š›”‘‹†• ‘ˆ Pten
—–ƒ– ‹…‡ ƒŽ•‘ ”‡•—Ž–• ‹ ‹…”‡ƒ•‡† •—……‹ƒ–‡ Ž‡˜‡Ž• ȋ–‹…‘ ‡– ƒŽǤǡ ƒ—•…”‹’– ‹
’”‡’ƒ”ƒ–‹‘ȌǤ •‹‹Žƒ” ’”‘Ǧ–”ƒ•ˆ‘”‹‰ …‘–”‹„—–‹‘ ‹‰Š– ƒŽ•‘ –ƒ‡ ’Žƒ…‡ ‹
‡†‘‡–”‹ƒŽ‡’‹–Š‡Ž‹ƒŽ…‡ŽŽ•‹•’‘”ƒ†‹…–›’‡ȋ‡†‘‡–”‹‘‹†Ȍ‡†‘‡–”‹ƒŽ…ƒ…‡”ǡ™Š‡”‡
Ž‘••‘ˆ‹•–Š‡‡ƒ”Ž‹‡•–”‡…‘‰‹œ‡†‰‡‡–‹…ƒŽ–‡”ƒ–‹‘ȋͷͲȌǤ
‹…‡–Š‡ƒƒŽ›•‹•‘ˆ‡ƒ”Ž›Š›’‡”’Žƒ•–‹…Ž‡•‹‘•™‹–Š͵ƒ…–‹˜ƒ–‹‘‹Š—ƒ–‹••—‡•‹•
‘–‡ƒ•‹Ž›ˆ‡ƒ•‹„Ž‡ǡƒ†”‡Ž‡˜ƒ–‡š’”‡••‹‘†ƒ–ƒ•‡–•ƒ”‡‘–’—„Ž‹…Ž›ƒ˜ƒ‹Žƒ„Ž‡ǡ–‘‡š–‡†
‘—” ‘†‡Ž –‘ ƒ††‹–‹‘ƒŽ Š—ƒ –‹••—‡• ™‡ —•‡† –Š‡ …‘‹‡ †ƒ–ƒ„ƒ•‡ǡ ƒ† ˆ‘—†
•‡˜‡”ƒŽ†ƒ–ƒ•‡–•‹™Š‹…ŠPTEN†‘™Ǧ”‡‰—Žƒ–‹‘…‘‡š‹•–•™‹–Š‰Ž‘„ƒŽ”‡’”‡••‹‘‘ˆ
…›…Ž‡ ‰‡‡•Ǥ –‡”‡•–‹‰Ž›ǡ ‘‡ ‘ˆ –Š‡•‡ †ƒ–ƒ•‡–• ‹• †‡”‹˜‡† ˆ”‘ ‡ƒ”Ž› •–ƒ‰‡ …‘Ž‘”‡…–ƒŽ
…ƒ…‡” ȋͷͳȌǡ •—’’‘”–‹‰ ‘—” Š›’‘–Š‡•‹• –Šƒ– –Š‡•‡ ‡–ƒ„‘Ž‹… …Šƒ‰‡• ƒ› –ƒ‡ ’Žƒ…‡
™‡ŽŽ„‡ˆ‘”‡ˆ—ŽŽ‡‘’Žƒ•–‹…–”ƒ•ˆ‘”ƒ–‹‘Ǥ‹ƒŽŽ›ǡƒ”‡…‡–’”‘–‡‘‹…ƒ†‡–ƒ„‘Ž‘‹…
•–—†› ‘ˆ ‰ƒ•–”‹… …ƒ…‡”ǡ ƒ –—‘” –›’‡ ‘ˆ–‡ ƒ••‘…‹ƒ–‡† ™‹–Š ͵ ƒ…–‹˜ƒ–‹‘ǡ ˆ‘—†
‡š–‡•‹˜‡”‡’”‡••‹‘‘ˆ…›…Ž‡‰‡‡•ȋͷʹȌǤ
 …‘…Ž—•‹‘ǡ ‘—” …Šƒ”ƒ…–‡”‹œƒ–‹‘ ‘ˆ ƒ in vivo ‘†‡Ž ‘ˆ ͵ ƒ…–‹˜ƒ–‹‘ ‹ –Š›”‘‹†
‡’‹–Š‡Ž‹ƒŽ…‡ŽŽ•Šƒ•Ž‡†–‘–Š‡†‹•…‘˜‡”›‘ˆƒ‘˜‡Ž’ƒ–Š™ƒ›ǡ‹™Š‹…Š–Š‡‰Ž›…‘Ž›–‹…•™‹–…Š
‹• ‘– ƒ…Š‹‡˜‡† –Š”‘—‰Š †‹”‡…– —’Ǧ”‡‰—Žƒ–‹‘ ‘ˆ ‰Ž›…‘Ž›–‹… ‡œ›‡ ‡š’”‡••‹‘ ƒ†
ƒ…–‹˜‹–›ǡ„—–”ƒ–Š‡”–Š”‘—‰Š–Š‡ƒ…–‹˜‡‹Š‹„‹–‹‘‘ˆƒ†–Š‡…‘•‡“—‡–”‡’”‡••‹‘
ʹ͸
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
‘ˆ –Š‡ ‡š’”‡••‹‘ ‘ˆ …›…Ž‡ ƒ† ‰‡‡•ǡ Ž‡ƒ†‹‰ –‘ ‹’ƒ‹”‡– ‘ˆ
‹–‘…Š‘†”‹ƒŽ‡–ƒ„‘Ž‹•Ǥ
ʹ͹
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ACKNOWLEDGEMENTS
Š‹•™‘”™ƒ••—’’‘”–‡†„›–Š‡Ž„‡”–‹•–‡‹ƒ…‡”‡–‡”‘”‡
”ƒ–ǡƒ†„›
‰”ƒ–• –‘ ȋͳʹͺͻͶ͵ ƒ† ͳ͸͹ͺ͵ͻȌǤ ‹• ƒ ”‡…‹’‹‡– ‘ˆ –Š‡ ”ƒ Ǥ ‹”•…ŠŽ
ƒ”‡‡”…‹‡–‹•–™ƒ”†Ǥǡƒ†Ȁƒ••ƒ›•™‡”‡‡•–ƒ„Ž‹•Š‡†‹–Š‡–ƒ„Ž‡
•‘–‘’‡ ƒ† ‡–ƒ„‘Ž‘‹…• ‘”‡ ‘ˆ –Š‡ ‹ƒ„‡–‡• ‡•‡ƒ”…Š ƒ† ”ƒ‹‹‰ ‡–‡” ȋȌǡ
•—’’‘”–‡†„›‰”ƒ–͸ͲͲʹͲͷͶͳǤ
ʹͺ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
REFERENCES
ͳǤ ”‹–œǡƒŒƒ•Ǥ‡–ƒ„‘Ž‹•ƒ†’”‘Ž‹ˆ‡”ƒ–‹‘•Šƒ”‡…‘‘”‡‰—Žƒ–‘”›’ƒ–Š™ƒ›•
‹…ƒ…‡”…‡ŽŽ•Ǥ…‘‰‡‡ǤʹͲͳͲǢʹͻǣͶ͵͸ͻǦ͹͹Ǥ
ʹǤ ƒ”„—”‰Ǥ–Š‡‘”‹‰‹‘ˆ…ƒ…‡”…‡ŽŽ•Ǥ…‹‡…‡Ǥͳͻͷ͸Ǣͳʹ͵ǣ͵ͲͻǦͳͶǤ
͵Ǥ ”–‡‰ƒ ǡ ƒ…Š‡œǦ”ƒ‰‘ ǡ ‹‡”Ǧƒ…Š‡œ ǡ ƒ…Š‡œǦ‡‹œ‘ ǡ ‹ŽŽ‡”• ǡ
—‡œ˜ƒǤ
Ž—…‘•‡ƒ˜‹†‹–›‘ˆ…ƒ”…‹‘ƒ•Ǥƒ…‡”Ž‡––‡”•ǤʹͲͲͻǢʹ͹͸ǣͳʹͷǦ͵ͷǤ
ͶǤ Ž•–”‘ ǡ ƒ—‡” ǡ —œœƒ‹ ǡ ƒ”ƒ—•ƒ• ǡ ƒ””‹• ǡ Žƒ• ǡ ‡– ƒŽǤ –
•–‹—Žƒ–‡•ƒ‡”‘„‹…‰Ž›…‘Ž›•‹•‹…ƒ…‡”…‡ŽŽ•Ǥƒ…‡””‡•‡ƒ”…ŠǤʹͲͲͶǢ͸Ͷǣ͵ͺͻʹǦͻǤ
ͷǤ ‡˜‹‡ ǡ —œ‹‘Ǧ—–‡” Ǥ Š‡ …‘–”‘Ž ‘ˆ –Š‡ ‡–ƒ„‘Ž‹… •™‹–…Š ‹ …ƒ…‡”• „›
‘…‘‰‡‡•ƒ†–—‘”•—’’”‡••‘”‰‡‡•Ǥ…‹‡…‡ǤʹͲͳͲǢ͵͵Ͳǣͳ͵ͶͲǦͶǤ
͸Ǥ —˜‡Žǡ‡…‹‡•ǡ‡‘ǡƒƒǡ‹’‘˜•›ǡ‘—œƒǡ‡–ƒŽǤ…–‹˜ƒ–‹‘‘ˆƒ
‡–ƒ„‘Ž‹… ‰‡‡ ”‡‰—Žƒ–‘”› ‡–™‘” †‘™•–”‡ƒ ‘ˆ  …‘’Ž‡š ͳǤ ‘Ž‡…—Žƒ” …‡ŽŽǤ
ʹͲͳͲǢ͵ͻǣͳ͹ͳǦͺ͵Ǥ
͹Ǥ ‹‰ ǡ —Ž‹ ǡ ƒ›‡• ǡ ƒ™‡Ž ǡ ƒ”–‹‡œ ǡ ƒ”‹• ǡ ‡– ƒŽǤ ‘„‹ƒ–‘”‹ƒŽ
”‡‰—Žƒ–‹‘‘ˆ‡—”‘„Žƒ•–‘ƒ–—‘”’”‘‰”‡••‹‘„›Ǧ›…ƒ†Š›’‘š‹ƒ‹†—…‹„Ž‡ˆƒ…–‘”
ǦͳƒŽ’ŠƒǤƒ…‡””‡•‡ƒ”…ŠǤʹͲͳͲǢ͹ͲǣͳͲ͵ͷͳǦ͸ͳǤ
ͺǤ ‘’’‡‘Ž ǡ ‘—†• ǡ ƒ‰ Ǥ ––‘ ƒ”„—”‰̵• …‘–”‹„—–‹‘• –‘ …—””‡–
…‘…‡’–•‘ˆ…ƒ…‡”‡–ƒ„‘Ž‹•Ǥƒ–—”‡”‡˜‹‡™•ƒ…‡”ǤʹͲͳͳǢͳͳǣ͵ʹͷǦ͵͹Ǥ
ͻǤ •”ƒ‡Žǡ…Š™ƒ”–œǤŠ‡‡–ƒ„‘Ž‹…ƒ†˜ƒ–ƒ‰‡‘ˆ–—‘”…‡ŽŽ•Ǥ‘Ž‡…—Žƒ”…ƒ…‡”Ǥ
ʹͲͳͳǢͳͲǣ͹ͲǤ
ͳͲǤ ƒ‹—†‘ ǡ ƒ›•ƒŽ ǡ Šƒ†‡Ž Ǥ ‡˜‹•‹–‹‰ –Š‡ …›…Ž‡ǣ •‹‰ƒŽ‹‰ –‘ –—‘”
ˆ‘”ƒ–‹‘Ǥ”‡†•‹‘Ž‡…—Žƒ”‡†‹…‹‡ǤʹͲͳͳǢͳ͹ǣ͸ͶͳǦͻǤ
ʹͻ
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ͳͳǤ ‡ƒ‰‡” ǡ Ž‡‹Ǧœƒ–‘ ǡ ‹—”ƒ ǡ ‹ ”‹•–‘ˆƒ‘ Ǥ –‡ Ž‘•• ‹ –Š‡ ‘—•‡
–Š›”‘‹† …ƒ—•‡• ‰‘‹–‡” ƒ† ˆ‘ŽŽ‹…—Žƒ” ƒ†‡‘ƒ•ǣ ‹•‹‰Š–• ‹–‘ –Š›”‘‹† ˆ—…–‹‘ ƒ†
‘™†‡†‹•‡ƒ•‡’ƒ–Š‘‰‡‡•‹•Ǥƒ…‡”‡•ǤʹͲͲ͹Ǣ͸͹ǣͻͷͻǦ͸͸Ǥ
ͳʹǤ –‹…‘Ǧ”…‹—…Š ǡ ‹ƒ ǡ ‹ƒ‘ ǡ ‡ˆ‡–‘ˆˆ ǡ ‹ ”‹•–‘ˆƒ‘ Ǥ ”‘••Ǧ–ƒŽ
„‡–™‡‡ ͵ ƒ† ‡•–”‘‰‡ ‹ –Š‡ ‘—•‡ –Š›”‘‹† ’”‡†‹•’‘•‡• –‘ –Š‡ †‡˜‡Ž‘’‡– ‘ˆ
ˆ‘ŽŽ‹…—Žƒ”…ƒ”…‹‘ƒ•™‹–ŠƒŠ‹‰Š‡”‹…‹†‡…‡‹ˆ‡ƒŽ‡•Ǥ…‘‰‡‡ǤʹͲͳͲǢʹͻǣͷ͸͹ͺǦͺ͸Ǥ
ͳ͵Ǥ ‹ŽŽ‡”ǡ‡ƒ‰‡”ǡƒ‡”ǡ‹ƒ‘ǡ‡ˆ‡–‘ˆˆǡ‹”‹•–‘ˆƒ‘Ǥ…‘‰‡‹…”ƒ•
”‡“—‹”‡• •‹—Ž–ƒ‡‘—• ͵ •‹‰ƒŽ‹‰ –‘ ‹†—…‡ ƒ…–‹˜ƒ–‹‘ ƒ† –”ƒ•ˆ‘” –Š›”‘‹†
‡’‹–Š‡Ž‹ƒŽ…‡ŽŽ•‹˜‹˜‘Ǥƒ…‡”‡•ǤʹͲͲͻǢ͸ͻǣ͵͸ͺͻǦͻͶǤ
ͳͶǤ –‹…‘ ”…‹—…Š ǡ —••‘ ǡ ‹ƒ ǡ ƒ‰ ǡ ƒ•”ƒ–Š ǡ ‹ƒ‘ ǡ ‡– ƒŽǤ
Š›”‘…›–‡Ǧ•’‡…‹ˆ‹…‹ƒ…–‹˜ƒ–‹‘‘ˆ’ͷ͵ƒ†–‡”‡•—Ž–•‹ƒƒ’Žƒ•–‹…–Š›”‘‹†…ƒ”…‹‘ƒ•
ˆƒ‹–Šˆ—ŽŽ›”‡…ƒ’‹–—Žƒ–‹‰Š—ƒ–—‘”•Ǥ…‘–ƒ”‰‡–ǤʹͲͳͳǢʹǤ
ͳͷǤ ƒ™Ž‘” ǡ ‘”ƒ ǡ •Š„› ǡ ‹ŽŽ‹ƒ• ǡ —””ƒ›Ǧƒ‹– ǡ ƒŽ‘‡ ǡ ‡– ƒŽǤ
••‡–‹ƒŽ ”‘Ž‡ ‘ˆ ͳ ‹ ”‡‰—Žƒ–‹‰ …‡ŽŽ •‹œ‡ ƒ† †‡˜‡Ž‘’‡– ‹ ‹…‡Ǥ „‘ Ǥ
ʹͲͲʹǢʹͳǣ͵͹ʹͺǦ͵ͺǤ
ͳ͸Ǥ ƒ–‡Žǡ‹ǡ‹š‘ǡŽƒ‰‘‹ǡ‡‡Š‘Žœ‡”ǡŠ‡ǡ‡–ƒŽǤ‡ƒ”…Šƒ„Ž‡‹‰ŠǦ
‡•‘Ž—–‹‘ʹ
‡Ž”‘–‡‘‡‘ˆ–Š‡—ƒ‘Ž‘”›’–Ǥ”‘–‡‘‡‡•ǤʹͲͲ͹Ǥ
ͳ͹Ǥ ‡ǡƒ–‡Žǡ‹—ǡ‹ǡƒŽ—•œƒǡ‘ˆˆƒǡ‡–ƒŽǤ”‘–‡‘‹…ƒƒŽ›•‡•‘ˆ
’ƒ…”‡ƒ–‹……›•–ˆŽ—‹†•Ǥƒ…”‡ƒ•ǤʹͲͲͻǢ͵ͺǣ‡͵͵ǦͶʹǤ
ͳͺǤ ‘ƒ”† ǡ —”ƒ† ǡ ‡›”‘Ž ǡ Šƒ•‡ƒ—‡ ǡ Šƒ—˜‹ ǡ ‘”‹‘ ǡ ‡– ƒŽǤ
‹–‘…Š‘†”‹ƒŽ †›•ˆ—…–‹‘ ”‡•—Ž–• ˆ”‘ ‘š‹†ƒ–‹˜‡ •–”‡•• ‹ –Š‡ •‡Ž‡–ƒŽ —•…Ž‡ ‘ˆ †‹‡–Ǧ
‹†—…‡†‹•—Ž‹Ǧ”‡•‹•–ƒ–‹…‡ǤŽ‹˜‡•–ǤʹͲͲͺǢͳͳͺǣ͹ͺͻǦͺͲͲǤ
͵Ͳ
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ͳͻǤ ƒ‹”• ǡ ƒ””‹• ǡ ƒ Ǥ ‡‰—Žƒ–‹‘ ‘ˆ …ƒ…‡” …‡ŽŽ ‡–ƒ„‘Ž‹•Ǥ ƒ–—”‡
”‡˜‹‡™•ǤʹͲͳͳǢͳͳǣͺͷǦͻͷǤ
ʹͲǤ ‡ƒ”…‡ǡ‘ƒ†‡”ǡŽ‡••‹ǤŠ‡—–•ƒ†„‘Ž–•‘ˆ
’”‘–‡‹‹ƒ•‡•Ǥƒ–
‡˜‘Ž‡ŽŽ‹‘ŽǤʹͲͳͲǢͳͳǣͻǦʹʹǤ
ʹͳǤ ƒ›ƒ•…ƒ•ǡƒƒ‘–‘ǡ”‹–ǡ”–Š—”ǡŽ‡••‹Ǥ˜ƒŽ—ƒ–‹‘‘ˆƒ’’”‘ƒ…Š‡•
–‘‰‡‡”ƒ–‹‘‘ˆ–‹••—‡Ǧ•’‡…‹ˆ‹…‘…Ǧ‹‹…‡Ǥ‹‘ŽŠ‡ǤʹͲͲ͸Ǣʹͺͳǣʹͺ͹͹ʹǦͺͳǤ
ʹʹǤ ‡ƒ‰‡” ǡ ”‡™‡” ǡ ƒ‹ ǡ — ǡ ‹ ”‹•–‘ˆƒ‘ Ǥ ƒƒŽ‹ƒ –ƒ”‰‡– ‘ˆ
”ƒ’ƒ›…‹‹•–Š‡‡›‡ˆˆ‡…–‘”‘ˆ’Š‘•’Šƒ–‹†›Ž‹‘•‹–‘ŽǦ͵ǦǦ‹‹–‹ƒ–‡†’”‘Ž‹ˆ‡”ƒ–‹˜‡•‹‰ƒŽ•
‹–Š‡–Š›”‘‹†ˆ‘ŽŽ‹…—Žƒ”‡’‹–Š‡Ž‹—Ǥƒ…‡”‡•ǤʹͲͲͺǢ͸ͺǣͶͶͶǦͻǤ
ʹ͵Ǥ —ǡŠ‡ǡƒǡ‡‰ǡƒ‰ǡŠƒǡ‡–ƒŽǤƒƒŽ‹ƒ–ƒ”‰‡–‘ˆ”ƒ’ƒ›…‹
—’Ǧ”‡‰—Žƒ–‹‘‘ˆ’›”—˜ƒ–‡‹ƒ•‡‹•‘‡œ›‡–›’‡ʹ‹•…”‹–‹…ƒŽˆ‘”ƒ‡”‘„‹…‰Ž›…‘Ž›•‹•ƒ†
–—‘”‰”‘™–ŠǤ”‘…ƒ–Ž…ƒ†…‹ǤʹͲͳͳǢͳͲͺǣͶͳʹͻǦ͵ͶǤ
ʹͶǤ ƒ”ƒ”ǡƒǡ‘™‡•ǡ—ǡ‘‡”ǡŽƒ›‹…ǡ‡–ƒŽǤš‡”…‹•‡ƒ†
ǦͳƒŽ’ŠƒǦ‹†‡’‡†‡–•›…Š”‘‹œƒ–‹‘‘ˆ–›’‡—•…Ž‡‡–ƒ„‘Ž‹•ƒ†˜ƒ•…—Žƒ–—”‡
„›‰ƒƒǤ‡ŽŽ‡–ƒ„ǤʹͲͳͳǢͳ͵ǣʹͺ͵Ǧͻ͵Ǥ
ʹͷǤ ƒ‰‡” ǡ ƒ†•…Š‹ ǡ –Ǧ‹‡””‡ ǡ ’‹‡‰‡Žƒ Ǥ Ǧƒ…–‹˜ƒ–‡† ’”‘–‡‹ ‹ƒ•‡
ȋȌ ƒ…–‹‘ ‹ •‡Ž‡–ƒŽ —•…Ž‡ ˜‹ƒ †‹”‡…– ’Š‘•’Š‘”›Žƒ–‹‘ ‘ˆ ǦͳƒŽ’ŠƒǤ ”‘… ƒ–Ž
…ƒ†…‹ǤʹͲͲ͹ǢͳͲͶǣͳʹͲͳ͹ǦʹʹǤ
ʹ͸Ǥ ‹‰—‡”‡Ǥ”ƒ•…”‹’–‹‘ƒŽ…‘–”‘Ž‘ˆ‡‡”‰›Š‘‡‘•–ƒ•‹•„›–Š‡‡•–”‘‰‡Ǧ”‡Žƒ–‡†
”‡…‡’–‘”•Ǥ†‘…”‡˜ǤʹͲͲͺǢʹͻǣ͸͹͹Ǧͻ͸Ǥ
ʹ͹Ǥ ƒ‘—”‹ ǡ ‡†„—”› ǡ ”‡––‘ ǡ —‰Š‡• ǡ ”‹ˆˆ‹ ǡ ƒŽŽƒ• ǡ ‡– ƒŽǤ
Šƒ…‡† Š‡’ƒ–‹–‹• ˜‹”—• ‰‡‘‡ ”‡’Ž‹…ƒ–‹‘ ƒ† Ž‹’‹† ƒ……——Žƒ–‹‘ ‡†‹ƒ–‡† „›
͵ͳ
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
‹Š‹„‹–‹‘‘ˆǦƒ…–‹˜ƒ–‡†’”‘–‡‹‹ƒ•‡Ǥ”‘…ƒ–Ž…ƒ†…‹ ǤʹͲͳͲǢͳͲ͹ǣͳͳͷͶͻǦ
ͷͶǤ
ʹͺǤ ‘”ƒǡ‡”–‘‡ǡ‡ƒ–Šǡ‡—ƒǡ‘—–‘ǡ‘‘†•ǡ‡–ƒŽǤ•—Ž‹
ƒ–ƒ‰‘‹œ‡•‹•…Š‡‹ƒǦ‹†—…‡†Š”ͳ͹ʹ’Š‘•’Š‘”›Žƒ–‹‘‘ˆǦƒ…–‹˜ƒ–‡†’”‘–‡‹‹ƒ•‡
ƒŽ’ŠƒǦ•—„—‹–• ‹ Š‡ƒ”– ˜‹ƒ Š‹‡”ƒ”…Š‹…ƒŽ ’Š‘•’Š‘”›Žƒ–‹‘ ‘ˆ ‡”ͶͺͷȀͶͻͳǤ ‹‘Ž Š‡Ǥ
ʹͲͲ͸Ǣʹͺͳǣͷ͵͵ͷǦͶͲǤ
ʹͻǤ —”Ž‡› ǡ ƒ””‡ ǡ ‘‘† ǡ †‡”•‘ ǡ ‡’ ǡ ‡ƒ• ǡ ‡– ƒŽǤ
‡‰—Žƒ–‹‘‘ˆǦƒ…–‹˜ƒ–‡†’”‘–‡‹‹ƒ•‡„›—Ž–‹•‹–‡’Š‘•’Š‘”›Žƒ–‹‘‹”‡•’‘•‡–‘
ƒ‰‡–•–Šƒ–‡Ž‡˜ƒ–‡…‡ŽŽ—Žƒ”…Ǥ‘—”ƒŽ‘ˆ‹‘Ž‘‰‹…ƒŽŠ‡‹•–”›ǤʹͲͲ͸Ǣʹͺͳǣ͵͸͸͸ʹǦ͹ʹǤ
͵ͲǤ Œ‘—†‡” ǡ —‡” ǡ —–‡” ǡ ƒŽ˜‹‘‹ ǡ ŠƒŽ‹ ǡ …Š‘Žœ ǡ ‡– ƒŽǤ ’Š‘•’Š‘”›Žƒ–‡• ƒ† ‹ƒ…–‹˜ƒ–‡• ƒŽ’Šƒ –‘ ’”‘‘–‡ ‡ˆˆ‹…‹‡– Ž‹’‘Ž›•‹•Ǥ Ǥ
ʹͲͳͲǢʹͻǣͶ͸ͻǦͺͳǤ
͵ͳǤ ‘„‡› ǡ ƒ› Ǥ • – –Š‡ ̶ƒ”„—”‰ ‹ƒ•‡̶ǫǦ–Ǧ‡‡”‰› ‡–ƒ„‘Ž‹•
‹–‡”ƒ…–‹‘•ƒ†‘…‘‰‡‡•‹•Ǥ‡‹ƒ…‡”‹‘ŽǤʹͲͲͻǢͳͻǣʹͷǦ͵ͳǤ
͵ʹǤ ‹ ǡ ƒ‘ ǡ ‹— ǡ ‡‡œƒ ǡ ƒ‰ Ǥ ›’‘š‹ƒǦ‹†—…‹„Ž‡ ˆƒ…–‘” ͳ ƒ†
†›•”‡‰—Žƒ–‡† …Ǧ›… …‘‘’‡”ƒ–‹˜‡Ž› ‹†—…‡ ˜ƒ•…—Žƒ” ‡†‘–Š‡Ž‹ƒŽ ‰”‘™–Š ˆƒ…–‘” ƒ†
‡–ƒ„‘Ž‹… •™‹–…Š‡• Š‡š‘‹ƒ•‡ ʹ ƒ† ’›”—˜ƒ–‡ †‡Š›†”‘‰‡ƒ•‡ ‹ƒ•‡ ͳǤ ‘Ž ‡ŽŽ ‹‘ŽǤ
ʹͲͲ͹Ǣʹ͹ǣ͹͵ͺͳǦͻ͵Ǥ
͵͵Ǥ ‡’”‡œ ǡ ‡”–‘‡ ǡ Ž‡••‹ ǡ —‡ ǡ ‹†‡” Ǥ Š‘•’Š‘”›Žƒ–‹‘ ƒ†
ƒ…–‹˜ƒ–‹‘ ‘ˆ Š‡ƒ”– ͸Ǧ’Š‘•’Š‘ˆ”—…–‘ǦʹǦ‹ƒ•‡ „› ’”‘–‡‹ ‹ƒ•‡ ƒ† ‘–Š‡” ’”‘–‡‹
‹ƒ•‡• ‘ˆ –Š‡ ‹•—Ž‹ •‹‰ƒŽ‹‰ …ƒ•…ƒ†‡•Ǥ ‘—”ƒŽ ‘ˆ ‹‘Ž‘‰‹…ƒŽ Š‡‹•–”›Ǥ
ͳͻͻ͹Ǣʹ͹ʹǣͳ͹ʹ͸ͻǦ͹ͷǤ
͵ʹ
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
͵ͶǤ ƒ”–Š‡Ž ǡ ‹‘ ǡ ‹ƒ‘ ǡ ƒƒ–ƒ‹ ǡ ‹ ǡ Š‹–Ž‘… ǡ ”Ǥǡ ‡– ƒŽǤ ‡‰—Žƒ–‹‘ ‘ˆ
ͳ ‰‡‡ –”ƒ•…”‹’–‹‘ „› –Š‡ •‡”‹‡Ȁ–Š”‡‘‹‡ ‹ƒ•‡ –ͳǤ ‹‘Ž Š‡Ǥ
ͳͻͻͻǢʹ͹ͶǣʹͲʹͺͳǦ͸Ǥ
͵ͷǤ Šƒ ǡ ƒ‰ ǡ ƒ‰ ǡ ‡ ǡ ‹‰ ǡ — ǡ ‡– ƒŽǤ ƒ…–ƒ–‡ †‡Š›†”‘‰‡ƒ•‡ ‹•
…”‹–‹…ƒŽˆ‘”Š›’‡”ƒ…–‹˜‡Ǧ‡†‹ƒ–‡†–—‘”‹‰‡‡•‹•Ǥƒ…‡”‡•ǤʹͲͳͳǢ͹ͳǣͳ͵ǦͺǤ
͵͸Ǥ ‡‡œƒ ǡ ‹ƒ‰ ǡ ‡—‰ ǡ ƒ••ƒ–‹‘ ǡ ‘…‘”†‡– ǡ ƒ‹”‡ ǡ ‡– ƒŽǤ
›’‘š‹ƒ ”‡•’‘•‡ ‡Ž‡‡–• ‹ –Š‡ ƒŽ†‘Žƒ•‡ ǡ ‡‘Žƒ•‡ ͳǡ ƒ† Žƒ…–ƒ–‡ †‡Š›†”‘‰‡ƒ•‡ ‰‡‡ ’”‘‘–‡”• …‘–ƒ‹ ‡••‡–‹ƒŽ „‹†‹‰ •‹–‡• ˆ‘” Š›’‘š‹ƒǦ‹†—…‹„Ž‡ ˆƒ…–‘” ͳǤ ‹‘Ž
Š‡Ǥͳͻͻ͸Ǣʹ͹ͳǣ͵ʹͷʹͻǦ͵͹Ǥ
͵͹Ǥ —– ǡ ƒ†‡” ‡‹†‡ Ǥ ‡”‘„‹… ‰Ž›…‘Ž›•‹•ǣ ‡‡–‹‰ –Š‡ ‡–ƒ„‘Ž‹…
”‡“—‹”‡‡–• ‘ˆ …‡ŽŽ ’”‘Ž‹ˆ‡”ƒ–‹‘Ǥ —ƒŽ ”‡˜‹‡™ ‘ˆ …‡ŽŽ ƒ† †‡˜‡Ž‘’‡–ƒŽ „‹‘Ž‘‰›Ǥ
ʹͲͳͳǢʹ͹ǣͶͶͳǦ͸ͶǤ
͵ͺǤ ƒ‰ ǡ ‹ŽŽ‘ ǡ Š‹ ǡ ‹Žƒ•–ƒ ǡ ƒ”–‡” ǡ ‹‡Ž•–‡‹ ǡ ‡– ƒŽǤ Š‡
–”ƒ•…”‹’–‹‘ ˆƒ…–‘” ›… …‘–”‘Ž• ‡–ƒ„‘Ž‹… ”‡’”‘‰”ƒ‹‰ —’‘ Ž›’Š‘…›–‡
ƒ…–‹˜ƒ–‹‘Ǥ—‹–›ǤʹͲͳͳǢ͵ͷǣͺ͹ͳǦͺʹǤ
͵ͻǤ ‘”ƒ‹ ǡ ƒ‰ƒ‘ ǡ ”‘†ƒ ǡ ‹ƒ”‡––‹ ǡ •‹†‘”‘ Ǥ ‘•• ‘ˆ ‡š’”‡••‹‘ ‘ˆ –Š‡
‘…‘•—’’”‡••‘”
‹
–Š›”‘‹†
‹…‹†‡–ƒŽ‘ƒ•
ƒ••‘…‹ƒ–‡•
™‹–Š
ͳ
’Žƒ•ƒ‡„”ƒ‡‡š’”‡••‹‘Ǥƒ‹‡”˜ƒ‡†‹…ƒǤʹͲͳʹǢͷͶǣͷͻǦ͸͵Ǥ
ͶͲǤ ƒ”…‹ƒǦƒ‘ǡ‘‰ǡ‘„„•ǡƒ—”‡–
ǡ
‹‘”‰‹ǡ†‡‘‡”ǡ‡–ƒŽǤ›•–‡‹…
‡Ž‡˜ƒ–‹‘‘ˆ‹†—…‡•ƒ–—‘”Ǧ•—’’”‡••‹˜‡‡–ƒ„‘Ž‹…•–ƒ–‡Ǥ‡ŽŽǤʹͲͳʹǢͳͶͻǣͶͻǦ͸ʹǤ
ͶͳǤ ‹Šƒ›Ž‘˜ƒǡŠƒ™ǤŠ‡•‹‰ƒŽŽ‹‰’ƒ–Š™ƒ›…‘‘”†‹ƒ–‡•…‡ŽŽ‰”‘™–Šǡ
ƒ—–‘’Šƒ‰›ƒ†‡–ƒ„‘Ž‹•Ǥƒ–—”‡…‡ŽŽ„‹‘Ž‘‰›ǤʹͲͳͳǢͳ͵ǣͳͲͳ͸Ǧʹ͵Ǥ
͵͵
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ͶʹǤ ƒ”†‹‡ ǡ ‘•• ǡ ƒ™Ž‡› Ǥ ǣ ƒ —–”‹‡– ƒ† ‡‡”‰› •‡•‘” –Šƒ–
ƒ‹–ƒ‹•‡‡”‰›Š‘‡‘•–ƒ•‹•Ǥƒ–—”‡”‡˜‹‡™•‘Ž‡…—Žƒ”…‡ŽŽ„‹‘Ž‘‰›ǤʹͲͳʹǢͳ͵ǣʹͷͳǦ͸ʹǤ
Ͷ͵Ǥ ‹‰ ǡ ‹ ǡ Ž‡‘• Ǥ —’’”‡••‹‘ ‘ˆ ƒ…–‹˜ƒ–‹‘ ˜‹ƒ Ͷͺͷ
’Š‘•’Š‘”›Žƒ–‹‘ „› Ǧ †—”‹‰ Š›’‡”‰Ž›…‡‹ƒ ‹• ‡†‹ƒ–‡† „› ƒ…–‹˜ƒ–‹‘ ‹
˜ƒ•…—Žƒ”•‘‘–Š—•…Ž‡…‡ŽŽ•Ǥ†‘…”‹‘Ž‘‰›ǤʹͲͳͳǢͳͷʹǣ͵ͳͶ͵ǦͷͶǤ
ͶͶǤ ƒ‰‘ ǡ —” ǡ Š‡‰ ǡ ‡ŽŽ‡•–‡‹ ǡ ƒ–Ž‡› ǡ ƒŠ Ǥ ’͹Ͳ͸ ‹ƒ•‡
’Š‘•’Š‘”›Žƒ–‡•‘•‡”‹‡Ͷͻͳ–‘‡†‹ƒ–‡Ž‡’–‹̵•‡ˆˆ‡…–‘ˆ‘‘†‹–ƒ‡Ǥ‡ŽŽ‡–ƒ„Ǥ
ʹͲͳʹǢͳ͸ǣͳͲͶǦͳʹǤ
ͶͷǤ ƒ—„‡”– ǡ ‘‹Ž› ǡ œ”‡‹‰ ǡ ”‹•• ǡ ƒ„‘”•ƒ ǡ ‘‰ ǡ ‡– ƒŽǤ ‹• ƒ
‡‰ƒ–‹˜‡ ”‡‰—Žƒ–‘” ‘ˆ –Š‡ ƒ”„—”‰ ‡ˆˆ‡…– ƒ† •—’’”‡••‡• –—‘” ‰”‘™–Š ‹ ˜‹˜‘Ǥ ‡ŽŽ
‡–ƒ„ǤʹͲͳ͵Ǣͳ͹ǣͳͳ͵ǦʹͶǤ
Ͷ͸Ǥ Žƒ›‹… ǡ ‘†‘ ǡ ‹‡ ǡ ‡ ǡ —ˆ‘—” ǡ ‘™‡• ǡ ‡– ƒŽǤ ‰ƒƒ
†‹”‡…–•ƒ†ƒ‹–ƒ‹•–Š‡–”ƒ•‹–‹‘–‘‘š‹†ƒ–‹˜‡‡–ƒ„‘Ž‹•‹–Š‡’‘•–ƒ–ƒŽŠ‡ƒ”–Ǥ‡ŽŽ
‡–ƒ„ǤʹͲͲ͹Ǣ͸ǣͳ͵ǦʹͶǤ
Ͷ͹Ǥ …ƒ”’—ŽŽƒ Ǥ ‡–ƒ„‘Ž‹… …‘–”‘Ž ‘ˆ ‹–‘…Š‘†”‹ƒŽ „‹‘‰‡‡•‹• –Š”‘—‰Š –Š‡ Ǧͳ
ˆƒ‹Ž›”‡‰—Žƒ–‘”›‡–™‘”Ǥ‹‘…Š‹‹‘’Š›•…–ƒǤʹͲͳͳǢͳͺͳ͵ǣͳʹ͸ͻǦ͹ͺǤ
ͶͺǤ ‹…Š‡”ǡ‡””›ǡ—ˆ‘—”ǡ‡”–‘•ǡƒ”ǡ–Ǧ‹‡””‡ǡ‡–ƒŽǤ‹Ǧ͵͹ͺȋȗȌ
‡†‹ƒ–‡• ‡–ƒ„‘Ž‹… •Š‹ˆ– ‹ „”‡ƒ•– …ƒ…‡” …‡ŽŽ• ˜‹ƒ –Š‡ Ǧͳ„‡–ƒȀ‰ƒƒ
–”ƒ•…”‹’–‹‘ƒŽ’ƒ–Š™ƒ›Ǥ‡ŽŽ‡–ƒ„ǤʹͲͳͲǢͳʹǣ͵ͷʹǦ͸ͳǤ
ͶͻǤ ‘„‡”–ǡ‡•–‡”ǡ‘Ž‹‡ǡ‰ǤŽ‡˜ƒ–‡†’Žƒ•ƒ•—……‹ƒ–‡‹ǡǡ
ƒ† —–ƒ–‹‘Ǧ’‘•‹–‹˜‡ ‹†‹˜‹†—ƒŽ•Ǥ ‡‡–‹…• ‹ ‡†‹…‹‡ ǣ ‘ˆˆ‹…‹ƒŽ Œ‘—”ƒŽ ‘ˆ –Š‡
‡”‹…ƒ‘ŽŽ‡‰‡‘ˆ‡†‹…ƒŽ
‡‡–‹…•ǤʹͲͳʹǤ
͵Ͷ
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ͷͲǤ —––‡” ǡ ‹ ǡ ‹–œ‰‡”ƒŽ† ǡ — ǡ ƒƒ ǡ ‡‡• ǡ ‡– ƒŽǤ Ž–‡”‡† ‡š’”‡••‹‘ƒ•ƒ†‹ƒ‰‘•–‹…ƒ”‡”ˆ‘”–Š‡‡ƒ”Ž‹‡•–‡†‘‡–”‹ƒŽ’”‡…ƒ…‡”•Ǥƒ–Žƒ…‡”
•–ǤʹͲͲͲǢͻʹǣͻʹͶǦ͵ͲǤ
ͷͳǤ ‘‰ǡ‘™‡›ǡ—ǡ‘ŠǡŠ‡ƒŠǤ̵‡–ƒ•–ƒ•‹•Ǧ’”‘‡̵•‹‰ƒ–—”‡ˆ‘”
‡ƒ”Ž›Ǧ•–ƒ‰‡ ‹•ƒ–…ŠǦ”‡’ƒ‹” ’”‘ˆ‹…‹‡– •’‘”ƒ†‹… …‘Ž‘”‡…–ƒŽ …ƒ…‡” ’ƒ–‹‡–• ƒ† ‹–•
‹’Ž‹…ƒ–‹‘• ˆ‘” ’‘••‹„Ž‡ –Š‡”ƒ’‡—–‹…•Ǥ Ž‹‹…ƒŽ Ƭ ‡š’‡”‹‡–ƒŽ ‡–ƒ•–ƒ•‹•Ǥ ʹͲͳͲǢʹ͹ǣͺ͵Ǧ
ͻͲǤ
ͷʹǤ ƒ‹ǡŠƒ‘ǡ‹ǡ‡‰ǡƒ‰ǡŠ‡ǡ‡–ƒŽǤ…‘„‹‡†’”‘–‡‘‹…•ƒ†
‡–ƒ„‘Ž‘‹…•’”‘ˆ‹Ž‹‰‘ˆ‰ƒ•–”‹……ƒ”†‹ƒ…ƒ…‡””‡˜‡ƒŽ•…Šƒ”ƒ…–‡”‹•–‹…†›•”‡‰—Žƒ–‹‘•‹
‰Ž—…‘•‡‡–ƒ„‘Ž‹•Ǥ‘Ž‡…—Žƒ”Ƭ…‡ŽŽ—Žƒ”’”‘–‡‘‹…•ǣǤʹͲͳͲǢͻǣʹ͸ͳ͹ǦʹͺǤ
͵ͷ
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
Table 1Ǥ”‘–‡‘‹…‹†‡–‹ˆ‹…ƒ–‹‘‘ˆ†‹ˆˆ‡”‡–‹ƒŽŽ›”‡‰—Žƒ–‡†…›…Ž‡‡œ›‡•Ǥ
Swiss-Prot
Protein Name
Expression
Name
ID Method
Score
Coverage
(WT vs. -/-)
Queries
matched
—……‹ƒ–‡†‡Š›†”‘‰‡ƒ•‡
ʹǤ͹ͳ
Ǧ
͹Ͷ
ͳ͸
Ȁ
͵
•‘…‹–”ƒ–‡†‡Š›†”‘‰‡ƒ•‡
͵ͶǤͳ͵
Ǧ
͹ͳ
ʹͷ
Ȁ
ƒŽƒ–‡†‡Š›†”‘‰‡ƒ•‡
ʹǤʹ͹
Ǧ
ͳͲ͸
͵Ͳ
Ȁ
…‘‹–ƒ–‡Š›†”ƒ–ƒ•‡
‘Ž›
ȀȀ
͵ͺʹ
Ȁ
ͳͷ
‹Š›†”‘Ž‹’‘›Ž†‡Š›†”‘‰‡ƒ•‡
͵Ǥʹʹ
ȀȀ
ʹͺ͵
Ȁ
ͳͶ
›”—˜ƒ–‡†‡Š›†”‘‰‡ƒ•‡
͸ǤͲͲ
ȀȀ
ͳͷͲ
Ȁ
͸
‹–”ƒ–‡•›–Šƒ•‡
ͶǤͲͳ
ȀȀ
ͳ͵ʹ
Ȁ
ͻ
͵͸
Downloaded from cancerres.aacrjournals.org on July 31, 2017. © 2013 American Association for Cancer Research.
Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
FIGURE LEGENDS
Figure 1. Metabolic reprogramming in Ptenthyr-/- mice. ȋȌ‡Žƒ–‹˜‡‡š’”‡••‹‘‘ˆ–Š‡
‹†‹…ƒ–‡†‰‡‡•‹™‹Ž†–›’‡ƒ†—–ƒ––Š›”‘‹†•Ǥƒ”•”‡’”‡•‡–‡ƒά‘ˆ–”‹’Ž‹…ƒ–‡
‡ƒ•—”‡‡–•Ǥ •–‡”‹•• ‹†‹…ƒ–‡ •‹‰‹ˆ‹…ƒ– ȋδͲǤͲͷȌ †‹ˆˆ‡”‡…‡•Ǥ ȋȌ ‡•–‡” „Ž‘–
•Š‘™‹‰ †‘™Ǧ”‡‰—Žƒ–‹‘ ‘ˆ ”‡’”‡•‡–ƒ–‹˜‡ …›…Ž‡ ‡œ›‡• ‹ —–ƒ– ‰Žƒ†•Ǥ ȋȌ
…›…Ž‡†‹ƒ‰”ƒ•Š‘™‹‰–Š‡‰‡‡•†‘™Ǧ”‡‰—Žƒ–‡†‹Pten–Š›”ǦȀǦ–Š›”‘‹†•ǤŠ‡…‘Ž‘”
•…ƒŽ‡ ”‡ˆŽ‡…–• …Šƒ‰‡• …ƒŽ…—Žƒ–‡† ˆ”‘ –Š‡ ˆˆ›‡–”‹š †ƒ–ƒǤ ‡‡• …‹”…Ž‡† ‹ ”‡† Šƒ˜‡
„‡‡ ˜ƒŽ‹†ƒ–‡† „› “Ǥ ȋȌ ‡ƒ– ƒ’ •Š‘™‹‰ –Š‡ ”‡’”‡••‹‘ ‘ˆ …›…Ž‡ ‰‡‡• ‹
Pten–Š›”ǦȀǦ –Š›”‘‹†• ƒ† ‹ ˆ‘ŽŽ‹…—Žƒ” …ƒ”…‹‘ƒ• ȋȌ ƒ”‹•‹‰ ‹ ‘Ž†‡” ‹…‡Ǥ ȋȌ …‘–‡–Ǧ„ƒ•‡† ƒ••‡••‡– ‘ˆ ‹–‘…Š‘†”‹ƒ —„‡” ‹ ͵ ‘–ŠǦ‘Ž† ™‹Ž† –›’‡ ƒ†
—–ƒ– ‹…‡ ȋ–‘’ ’ƒ‡ŽȌǤ š’”‡••‹‘ Ž‡˜‡Ž• ‘ˆ •‡Ž‡…–‡† ‹–‘…Š‘†”‹ƒŽ ‰‡‡• ‹ –Š‡
–Š›”‘‹†• ‘ˆ ™‹Ž† –›’‡ ƒ† —–ƒ– ‹…‡ ȋ„‘––‘ ’ƒ‡ŽȌǤ ƒ”• ”‡’”‡•‡– ‡ƒ ά ‘ˆ
–”‹’Ž‹…ƒ–‡ ‡ƒ•—”‡‡–•Ǥ •–‡”‹•• ‹†‹…ƒ–‡ •‹‰‹ˆ‹…ƒ– ȋδͲǤͲͷȌ †‹ˆˆ‡”‡…‡•Ǥ ȋȌ
‹–‘…Š‘†”‹ƒŽ †ƒƒ‰‡ ȋ•™‡ŽŽ‹‰ǡ …Žƒ”‹ˆ‹…ƒ–‹‘ǡ …”‹•–ƒ‡ †‹•”—’–‹‘Ȍ ‹ PtenǦȀǦ ‰Žƒ†•
†‡–‡…–‡†„›–”ƒ•‹••‹‘‡Ž‡…–”‘‹…”‘•…‘’›Ǥȋ
Ȍš›‰‡…‘•—’–‹‘”ƒ–‡•ȋȌ‹
…‘–”‘Ž ƒ† —–ƒ– ’”‹ƒ”› –Š›”‘…›–‡• …‡ŽŽ• ‹ ”‡•’‘•‡ –‘ ͳρ‰ȀŽ Ž‹‰‘›…‹ǡ ͳ ρ
Ž—‘”‘Ǧ…ƒ”„‘›Ž…›ƒ‹†‡’Š‡›ŽŠ›†”ƒœ‘‡ȋȌ‘”ʹρ–‹›…‹Ϊʹρ‘–‡‘‡Ǥ
ȋȌ š–”ƒ…‡ŽŽ—Žƒ” ƒ…‹†‹ˆ‹…ƒ–‹‘ ”ƒ–‡ ȋȌ ‹ …‘–”‘Ž ƒ† —–ƒ– ’”‹ƒ”› –Š›”‘…›–‡•
…‡ŽŽ•Ǥ α ͲǤͲͲͶȌǤ ȋȌ ‹–‘…Š‘†”‹ƒŽ ‡„”ƒ‡ ’‘Žƒ”‹œƒ–‹‘ ™ƒ• ‡ƒ•—”‡† ‹ ’”‹ƒ”›
–Š›”‘…›–‡•„›ˆŽ‘™…›–‘‡–”›—•‹‰Ǥ’”‡–”‡ƒ–‡–‘ˆ™–…‡ŽŽ•™ƒ•—•‡†–‘
†‡–‡”‹‡–Š‡„ƒ•‡Ž‹‡Ǥ
Figure 2. Enhanced glycolysis in Pten-/- thyroids. ȋȌ ƒ…–ƒ–‡Ž‡˜‡Ž•‹–Š‡–Š›”‘‹†•‘ˆ
͵‘–ŠǦ‘Ž†™‹Ž†–›’‡ƒ†—–ƒ– ‹…‡Ǥƒ”•”‡’”‡•‡–‡ƒάȋαͶ’‡”’‘‘ŽȌǤȋȌ
͵͹
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
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ƒ…–ƒ–‡ ’”‘†—…–‹‘ ”ƒ–‡ ‹ ’”‹ƒ”› …—Ž–—”‡• ‘ˆ ™‹Ž† –›’‡ ƒ† —–ƒ– –Š›”‘…›–‡•Ǥ ȋȌ
ͳͺ
‹…”‘ƒƒŽ›•‹•
•Š‘™‹‰‹…”‡ƒ•‡†‰Ž—…‘•‡—’–ƒ‡‹͵‘–ŠǦ‘Ž†—–ƒ– ‹…‡
…‘’ƒ”‡† –‘ ™‹Ž† –›’‡ …‘–”‘Ž•Ǥ Š‡ ‹•‡– •Š‘™• ƒ –”ƒ•˜‡”•‡ •‡…–‹‘ …‡–‡”‡† ‘ –Š‡
–Š›”‘‹†ǤȋȌš’”‡••‹‘Ž‡˜‡Ž•‘ˆ–Š‡‹†‹…ƒ–‡†‰‡‡•‹™‹Ž†–›’‡ƒ†—–ƒ––Š›”‘‹†•Ǥ
ƒ”• ”‡’”‡•‡– ‡ƒ ά ‘ˆ –”‹’Ž‹…ƒ–‡ ‡š’‡”‹‡–•Ǥ ȋȌ ‡•–‡” „Ž‘– •Š‘™‹‰ ‘
†‡”‡‰—Žƒ–‹‘ ‘ˆ ‹ˆͳα ƒ† ʹ ‹ —–ƒ– ‰Žƒ†•Ǥ •–‡”‹•• ‹†‹…ƒ–‡ •‹‰‹ˆ‹…ƒ–
ȋδͲǤͲͷȌ†‹ˆˆ‡”‡…‡•Ǥ
Figure 3. TCA cycle/OXPHOS gene repression depends on Pdk1 but not mTOR.
ȋǡȌ‘••‘ˆPdk1 ”‡•–‘”‡•‘”ƒŽŽ‡˜‡Ž•‘ˆ…›…Ž‡ȋȌƒ†‹–‘…Š‘†”‹ƒŽȋȌ‰‡‡
‡š’”‡••‹‘‹Pten–Š›”ǦȀǦ‹…‡. ƒ”•”‡’”‡•‡–‡ƒά‘ˆ–”‹’Ž‹…ƒ–‡ƒ••ƒ›•Ǥ ȋȌ‘••‘ˆ
Pdk1 ”‡•–‘”‡•‘”ƒŽŽ‡˜‡Ž•‘ˆŽƒ…–ƒ–‡‹Pten–Š›”ǦȀǦ–Š›”‘‹†‰Žƒ†•Ǥƒ”•”‡’”‡•‡–‡ƒά
ȋαͶ’‡”‰‡‘–›’‡ȌǤ ȋȌ…Š‡‡‘ˆͲͲͳƒ†‹‹•–”ƒ–‹‘–‘͵Ǧ‘–Š‘Ž†‹…‡ƒ†
™‡•–‡” „Ž‘– ˜‡”‹ˆ‹…ƒ–‹‘ ‘ˆ ‡ˆˆ‡…–‹˜‡ ‹Š‹„‹–‹‘ ‘ˆ  ƒ…–‹˜‹–›Ǥ ȋȌ  ‹Š‹„‹–‹‘
•—’’”‡••‡•–Š›”‘…›–‡’”‘Ž‹ˆ‡”ƒ–‹‘Ǥƒ”•”‡’”‡•‡–‡ƒάȋαͷ’‡”‰‡‘–›’‡ȌǤȋǡ
Ȍ
 ‹Š‹„‹–‹‘ ˆƒ‹Ž• –‘ ”‡•–‘”‡ ‘”ƒŽ …›…Ž‡ ȋȌ ƒ† ‹–‘…Š‘†”‹ƒŽ ȋ
Ȍ ‰‡‡
‡š’”‡••‹‘Ǥƒ”•”‡’”‡•‡–‡ƒά‘ˆ–”‹’Ž‹…ƒ–‡ƒ••ƒ›•ǤȋȌ‹Š‹„‹–‹‘ˆƒ‹Ž•–‘
”‡•…—‡Žƒ…–ƒ–‡‹…”‡ƒ•‡‹—–ƒ–‹…‡Ǥƒ”•”‡’”‡•‡–‡ƒάȋαͶ’‡”‰‡‘–›’‡ȌǤ
•–‡”‹••‹†‹…ƒ–‡•‹‰‹ˆ‹…ƒ–ȋδͲǤͲͷȌ†‹ˆˆ‡”‡…‡•Ǥ
Figure 4. AMPK is repressed in Ptenthyr-/- thyroids. ȋȌ “ ’”‘ˆ‹Ž‹‰ ‘ˆ ‡–ƒ„‘Ž‹…
–”ƒ•…”‹’–‹‘ƒŽ ”‡‰—Žƒ–‘”• ‹ –Š‡ –Š›”‘‹†• ‘ˆ …‘–”‘Ž ƒ† —–ƒ– ‹…‡Ǥ ƒ”• ”‡’”‡•‡–
‡ƒά‘ˆ–”‹’Ž‹…ƒ–‡ƒ••ƒ›•Ǥ•–‡”‹••‹†‹…ƒ–‡•‹‰‹ˆ‹…ƒ–ȋδͲǤͲͷȌ†‹ˆˆ‡”‡…‡•ǤȋȌ
‡•–‡”„Ž‘––‹‰ƒƒŽ›•‹•‘ˆƒ…–‹˜ƒ–‹‘‹™‹Ž†–›’‡ƒ†—–ƒ–‰Žƒ†•ǤȋȌǡ
͵ͺ
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
ǡƒ†Ž‡˜‡Ž•‹–Š›”‘‹†•ˆ”‘™‹Ž†–›’‡ƒ†Pten–Š›”ǦȀǦ‹…‡Ǥƒ”•”‡’”‡•‡–‡ƒ
άȋα͵’‘‘Ž•‘ˆͳͲ–Š›”‘‹†•’‡”‰‡‘–›’‡ȌǤ
Figure 5. AICAR-mediated AMPK activation reverts the metabolic switch in
Ptenthyr-/- thyroids. ȋȌ‘’’ƒ‡Žǣ…Š‡‡‘ˆƒ†‹‹•–”ƒ–‹‘ǣ͵‘–ŠǦ‘Ž†‹…‡
™‡”‡ ‹Œ‡…–‡† ™‹–Š ȋͶͲͲ ‰Ȁ‰Ȁ†ƒ›Ȍ ˆ‘” Ͷ ™‡‡•Ǥ ‘––‘ ’ƒ‡Žǣ ‡•–‡”
„Ž‘––‹‰ ƒƒŽ›•‹• ‘ˆ –Š‡ ‡ˆˆ‡…– ‘ˆ in vivo –”‡ƒ–‡– ‘ –Š‡ ’Š‘•’Š‘”›Žƒ–‹‘ ‘ˆ
Ǥ ȋǡȌ š’”‡••‹‘ Ž‡˜‡Ž• ‘ˆ •‡Ž‡…–‡† ‡–ƒ„‘Ž‹… –”ƒ•…”‹’–‹‘ƒŽ ”‡‰—Žƒ–‘”• ȋȌ ƒ†
…›…Ž‡ ‰‡‡• ȋȌ —’‘ in vivo –”‡ƒ–‡–Ǥ ƒ”• ”‡’”‡•‡– ‡ƒ ά ‘ˆ
–”‹’Ž‹…ƒ–‡ ƒ••ƒ›•Ǥ ȋȌ ‡•–‡” „Ž‘––‹‰ •Š‘™‹‰ –Šƒ– in vivo –”‡ƒ–‡– ”‡•…—‡•
…›…Ž‡ ‡œ›‡• ‡š’”‡••‹‘Ǥ ȋȌ š’”‡••‹‘ Ž‡˜‡Ž• ‘ˆ ‹–‘…Š‘†”‹ƒŽ ‰‡‡• ‹ –Š‡
–Š›”‘‹†• ‘ˆ …‘–”‘Ž ƒ† Ǧ–”‡ƒ–‡† ‹…‡Ǥ ƒ”• ”‡’”‡•‡– ‡ƒ ά ‘ˆ –”‹’Ž‹…ƒ–‡
ƒ••ƒ›•Ǥ ȋȌ ”ƒ•‹••‹‘ ‡Ž‡…–”‘ ‹…”‘•…‘’› •Š‘™‹‰ ‹–‘…Š‘†”‹ƒ •–”—…–—”‡ ‹ –Š‡
–Š›”‘‹†• ‘ˆ ™‹Ž† –›’‡ǡ PtenǦȀǦ ƒ† Ǧ–”‡ƒ–‡† PtenǦȀǦ ‹…‡Ǥ ȋ
Ȍ ˆˆ‡…– ‘ˆ –Š‡ in vivo
–”‡ƒ–‡–‘Žƒ…–ƒ–‡’”‘†—…–‹‘Ǥƒ”•”‡’”‡•‡–‡ƒάȋαͶ’‡”‰‡‘–›’‡ȌǤ
ȋȌ Š›”‘‹† ™‡‹‰Š– ‹ ™‹Ž† –›’‡ǡ —–ƒ–ǡ ƒ† –”‡ƒ–‡† —–ƒ– ‹…‡Ǥ ȋȌ
”‘Ž‹ˆ‡”ƒ–‹‘‹†‡š‘ˆ–Š›”‘‹†•ˆ”‘™‹Ž†–›’‡ǡ—–ƒ–ǡƒ†–”‡ƒ–‡†—–ƒ–‹…‡
ƒ•‡ƒ•—”‡†„›”†‹…‘”’‘”ƒ–‹‘Ǥ•–‡”‹••‹†‹…ƒ–‡•‹‰‹ˆ‹…ƒ–ȋδͲǤͲͷȌ†‹ˆˆ‡”‡…‡•Ǥ
Figure 6. The AKT/AMPK/PGC1α/TCA cycle axis is conserved in human thyroid
cancer cells. ȋȌ ‡•–‡”„Ž‘––‹‰ƒƒŽ›•‹•‘ˆ–Š‡‡ˆˆ‡…–‘ˆ–”‡ƒ–‡–‘
’Š‘•’Š‘”›Žƒ–‹‘•–ƒ–—•‹Ǧͳ͵͵…‡ŽŽ•ǤȋȌš’”‡••‹‘ƒƒŽ›•‹•‘ˆ”‡’”‡•‡–ƒ–‹˜‡
…›…Ž‡ ‰‡‡• ‹ Ǧͳ͵͵ …‡ŽŽ• —’‘ –”‡ƒ–‡–Ǥ ƒ”• ”‡’”‡•‡– ‡ƒ ά ‘ˆ
–”‹’Ž‹…ƒ–‡ƒ••ƒ›•ǤȋȌǦ‹†—…‡†‹…”‡ƒ•‡‹–Š‡‡š’”‡••‹‘Ž‡˜‡Ž•‘ˆ”‡’”‡•‡–ƒ–‹˜‡
…›…Ž‡‰‡‡•‹Ǧͳ͵͵…‡ŽŽ•–”ƒ•ˆ‡…–‡†™‹–Š™‹Ž†–›’‡‘”—–ƒ–ȋȌ
ͳαǤ
͵ͻ
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Author Manuscript Published OnlineFirst on June 24, 2013; DOI: 10.1158/0008-5472.CAN-13-1429
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–Š‡–‘’ǡ™‡•–‡”„Ž‘––‹‰ƒƒŽ›•‹••Š‘™‹‰…‘’ƒ”ƒ„Ž‡‡š’”‡••‹‘Ž‡˜‡Ž•‘ˆ–”ƒ•ˆ‡…–‡†
™‹Ž†–›’‡ƒ†—–ƒ–ȋȌ
ͳαȋ•’‡…‹ˆ‹…„ƒ†ƒ”‡†™‹–Šƒƒ•–‡”‹•ȌǤȋȌ‡•–‡”
„Ž‘––‹‰ ƒƒŽ›•‹• ‘ˆ –Š‡ ‡ˆˆ‡…– ‘ˆ ͺͻǡ ͳʹͲǡ ƒ† –Š‡‹” …‘„‹ƒ–‹‘ ‘ ’Š‘•’Š‘”›Žƒ–‹‘ ‹ Ǧͳ͵͵ …‡ŽŽ•Ǥ ȋȌ š’”‡••‹‘ Ž‡˜‡Ž• ‘ˆ ”‡’”‡•‡–ƒ–‹˜‡ ‡–ƒ„‘Ž‹…
–”ƒ•…”‹’–‹‘ƒŽ ”‡‰—Žƒ–‘”• ƒ† …›…Ž‡ ‰‡‡• —’‘ ͺͻǡ ͳʹͲǡ ƒ† –Š‡‹”
…‘„‹ƒ–‹‘ –”‡ƒ–‡–‹Ǧͳ͵͵…‡ŽŽ•Ǥƒ”•”‡’”‡•‡–‡ƒά‘ˆ–”‹’Ž‹…ƒ–‡ƒ••ƒ›•Ǥ
ȋȌ—…‹ˆ‡”ƒ•‡ƒ••ƒ›•Š‘™‹‰
ͳα’”‘‘–‡”ƒ…–‹˜‹–›‹Ǧͳ͵͵…‡ŽŽ•ƒˆ–‡”–”‡ƒ–‡–
™‹–Š –Š‡ ‹†‹…ƒ–‡† ‹Š‹„‹–‘”•Ǥ ƒ”• ”‡’”‡•‡– ‡ƒ ά ȋα͵ ’‡” –”‡ƒ–‡–ȌǤ ȋ
Ȍ
‡•–‡”„Ž‘––‹‰ƒƒŽ›•‹•‘ˆ–Š‡‡ˆˆ‡…–‘ˆ–”‡ƒ–‡–‘ˆͳ͸ƒ†ͺͷͲͷ……‡ŽŽ•‘
’Š‘•’Š‘”›Žƒ–‹‘ •–ƒ–—•Ǥ ȋȌ ‡•–‡” „Ž‘––‹‰ ƒƒŽ›•‹• ‘ˆ –Š‡ ‡ˆˆ‡…– ‘ˆ ͺͻǡ
ͳʹͲƒ†–Š‡‹”…‘„‹ƒ–‹‘‘’Š‘•’Š‘”›Žƒ–‹‘•–ƒ–—•‹ͳ͸ƒ†ͺͷͲͷ…
…‡ŽŽ•Ǥ ȋȌ š’”‡••‹‘ Ž‡˜‡Ž• ‘ˆ –Š‡ ESRRG ƒ† PPARGC1B ‡–ƒ„‘Ž‹… –”ƒ•…”‹’–‹‘ƒŽ
”‡‰—Žƒ–‘”•—’‘ ͺͻǡͳʹͲǡ ƒ† –Š‡‹” …‘„‹ƒ–‹‘ –”‡ƒ–‡– ‹ ͳ͸ ƒ† ͺͷͲͷ…
…‡ŽŽ•Ǥ ƒ”• ”‡’”‡•‡– ‡ƒ ά ‘ˆ –”‹’Ž‹…ƒ–‡ ƒ••ƒ›•Ǥ •–‡”‹•• ‹†‹…ƒ–‡ •‹‰‹ˆ‹…ƒ–
ȋδͲǤͲͷȌ†‹ˆˆ‡”‡…‡•ǤȋȌ”‘Ž‹ˆ‡”ƒ–‹‘‘ˆ‘—•‡ƒ†Š—ƒ–Š›”‘‹†…ƒ…‡”…‡ŽŽ•–”‡ƒ–‡†
™‹–Šͳƒ†…‘—–‡†ƒˆ–‡”͹ʹŠǤƒ”•”‡’”‡•‡–‡ƒά‘ˆ–”‹’Ž‹…ƒ–‡ƒ••ƒ›•Ǥ
ͶͲ
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Inhibition of AMPK and Krebs cycle gene expression drives
metabolic remodeling of Pten-deficient preneoplastic thyroid
cells
Valeria G Antico Arciuch, Marika A Russo, Kristy S Kang, et al.
Cancer Res Published OnlineFirst June 24, 2013.
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