相対発現量 (/ β-actin)

ϯ˨5˥ϰ˽̤Rĭɿ^ƒƹ̤

5-1. ǒͳ

1983ǎDJohnstoneŗ΀èή̲ɠΘ‡Ƀ^ƒ 100nmwmqƹ̤R˽

̤Qĭɿ[„ƒYl‡ʾ͕\ϨPan and Johnstone, 1983ϩDbqƹ̤r GExosomeHlţôW„dϨJohnstone et al., 1987ϩE\Q\DǛȹD Exosomer˽̤Qĭɿ[„ƒšÃ͎Ϩgarbage bagsϩl\jD˽̤ĦD

˽̤̲qĂĭn¥Ñµ•΀‡˽̤ƒoȓjƒǞļel̚O„jMdEǟ Ƙr¶©žq΁͈ʩǻʝΠ˝oPMjD̆ʣ΁͈ʩR¼Æ–Ï·ÑŠǻ ȹorǪ͔kLgd¬Ëџ¹ŽÌÑΡ»¥Ô‡˽̤ƒoȕĬ^ƒY l‡ʾ͕\DExosome rÚ͔ongd}q‡ȕĬ^ƒǞļRLƒĺͳ ôWdd|kLƒE\Q\DqfoExosome ršÃ͎krnUDɦ̦‡

ȍhƹ̤kLƒYlRğʸƣ‡ßǩoĭQgjSdϨRaposo et al., 1996, Bobrie et al., 2011ϩE[o Exosome or miRNA RŦy„ƒYl‡

2007 ǎo Valadi RƊŨ\dYl‡ƖɦoϨValadi et al., 2007ϩD ExosomeˎˢRʄɷiMdE

5-2. ˽̤ƒƹ̤qɫŜlūtţ

Johnstone o€ƒ Exosome qʾ͕r 1983 ǎkLƒRϨţôW„j qr1987ǎϩDb„÷ķoRonquistRProstasomelMN40K500nm wmqƔS[ķ΀èή̲ɠΘ‡Ƀ^ƒ˽̤ƒƹ̤‡ʾ͕\jMƒ

ϨRonquist and Hedström, 1977ϩEProstsomerķˤ̱Ø˄˽̤Qĭ ɿ[„DάǮÙoPWƒ˵ƟqČ͸lΟŁ̦ʦǤoƮÛ\jMƒlʨžr

̚O„jMƒEydDYq€Nn˽̤ƒƹ̤orɡFnūtţRôW

„jP‚DmicrovesicleDectosomeDmicroparticleDshedding vesicleD

b\j exosome lɡFnūtţRL‚DȹorŢ]}q‡Ȏ\jMƒY

lRLƒRDʰɍ^ƒ˽̤~˴ƟǠDĭɿ̀ΆnmqȽȸnƧ̗ôWkņ ij[„ƒYl}L‚Dϐnjoŷdz^ƒʨʣkLƒEbq̢ȻorɡFn–

ÍÔ»Rďijoˎˢ‡Λ|jSdYlRLƒE[oExosomelrRNA ĭ͛Ϋ˺͓ŠĀ‡Ȏ^ţ˛k}L‚DĢUijqʡlūtţRŢ]kLƒY l r ϐ nj o ŷ dz ^ ƒ E b Y k D2011 ǎ D Ȱ d o Ż ψ ƣ ù ISEV ϨInternational Society for Extracellular VesiclesϩR͡ˤ[„Dl‚L O_Extracellular vesicles Ϩ˽̤ƒƹ̤ϩlMNūtţk̃ՇŹ…N l\jMƒEðǢDɡFnƧ̗}Yqƣùkɼ|„DĀ˷Ņ[„ƒș́

ǮRLƒRDɉˎˢkrʨžȘƗ[„jMƒExtracellular vesiclesϨEVsϩ lMNūtţ‡Ąʮ\D100nmķǢqƹ̤‡Ȏ^Yll^ƒE

5-3. EVsqĭɿɦɠ

EVs qĭɿ̀Άr÷Ùq€NǹΆRȚŰ[„jMƒϨRaposo and Stoorvogel, 2013ϩEy_DÑ­›Œ¬ÔŸo€gjǜǻ[„dÑ­

£ Ô Ä q Ħ ĕ k Ĭ ̼ ^ ƒ € N o ǜ ǻ [ „ ƒ ̲ ƹ ̤ Dintraluminal membrane vesicle ϨILVϩ‡ƓȫŦ{multivesicular body ϨMVBϩR

˽̤̲l͇Š\j˽̤ƒuȩĬ[„ƒILVRExosomel[„jMƒE[

oķΒ\dmicrovesicleDshedding vesiclelūs„ƒϜMq}qr˽

̤̲QfT„ƒ€NoȩĬ[„ƒƹ̤l[„jMƒRDye͛ȶor̶

gjP_Dĭɿ̀ΆqȶˑnņĭRύ\UDĭɿɦɠqͪ\M͛ɐrð ǢqˎˢoɅǡ[„ƒE\Q\DMUhQĭɿoŶƟRƊŨ[„j MƒEÕhrDCa2+kL‚D΁ˀ͈ʺ˽̤ɕ K562 l̟ʑ˽̤oPMj

˽̤Ħq Ca2+ʗǕRƎľ^ƒl EVs qĭɿRǗS΃Y[„ƒYlRƊ

Ũ[„jMƒϨSavina et al., 2003, Raposo et al., 1997ϩE[oĭɿo rƓUqĭƟqξÛR˔ů[„jMƒEʢo ILV qǜǻoξÛ^ƒ

endosomal sorting complex required for transport ϨESCRTϩ͓ŠĀq

™ÑÁԱѬͅˀ΀~AlixnmREVsqĭɿoYlRƊŨ[„

jMƒϨGéminard et al., 2004, Baietti et al., 2012, Tamai et al., 2010ϩ

\Q\DYq ESCRT ͓ŠĀo€ƒ̀Άr EVs qĭɿoǪ͔nMlMN ƊŨ}LƒϨTrajkovic et al., 2008ϩEYqƊŨkŗ΀¡ËíʭʬΫ

˺neutral sphingomyelinase 2 ϨnSMase2ϩREVsqĭɿ‡ĵǥ^ƒ

Yl‡ȶQo\jMƒEóo}MUhQqGTPƑțΫ˺q Rab¹‰Ã ÌÔΧúƟRĭɿoξ†gjP‚DRab11DRab27aDRab27bDRab35 nmREVsqĭɿoξÛ\jMƒYlRƊŨ[„jMƒϨSavina et al., 2002, Ostrowski et al.,2010, Hsu et al., 2010ϩEYq€NoƓUqĭƟ~

ŶƟRξÛ^ƒYlk EVs rʭʬDĭɿ[„ƒYlRȶQongjS dRDĢƭ͛ȶor̶gjMnME

5-4. EVslmiRNA

ØΒ\d€NoEVsˎˢ‡[oʄɷiWdƊŨRDEVsqßomiRNA RŦy„ƒDlMN}qkLgdE2007ǎoValadirŸ̟ʑ˽̤

ɕMC/9l¶¬̟ʑ˽̤ɕHMC-1ʰɍq EVsoRNARŦy„YlD[

oŒ•ÏŠÎŒ‡ʮMjMC/9˽̤ʰɍEVso˸1300˞ϜqmRNA l121˞Ϝq miRNARŦy„ƒYl‡ƊŨ\dϨValadi et al., 2007ϩE ǟrEVsoŦy„ƒ RNAR˽̤ν‡˜Ł^ƒYl‡ͤȶ^ƒd|oD MC/9 ˽̤ɕʰɍq EVsDhy‚Ÿʰɍq EVs ‡¶¬q˽̤ɕkL

ƒHMC-1˽̤oʊľ\D¥Ñµ•΀qʾʨ‡èɨěώɷʂŁʀl΀ΰĭ

ɐʀk͛ɐ\dEbq́ɑDHMC-1 ˽̤ĦoŸq¥Ñµ•΀qơž Rͬ|„dEbqNf3hq¥Ñµ•΀ohMjrMC/9˽̤ɕʰɍq EVsorŦy„jMnMRDb„qmRNArŒ•ÏŠÎŒkɛĬ[

„jMd}qkLgdEhy‚DĆÛĕqMC/9˽̤ʰɍqmRNAREVs kΟs„jDŘƭĕq HMC-1 ˽̤Ħk̙͢[„dș́Ǯ‡˔\jMƒE ǟrGmRNAHq˽̤ν˜Łr˔\dRDGmiRNAHq˽̤νq˜Ł~

˜Ł\dmiRNA qɦ̦‡˔\jMnQgdE

EVsoŦy„ƒmiRNARGšÃHkLƒqQDb„l}˽̤ν‡˜Ł\D Řƭĕq˽̤kɦ̦^ƒ}qnqQDY„oξ\jr2010ǎDŢȹɅo 3ɉqͳȭRƊŨ[„dEy_DPegtelrEBV ϨEpstein-Barr Virusϩ oǶɒ\dÌѵ̼ʩɡ˽̤ɕ LCL l EBV oǶɒ\jMnMōʩʰɍ qɥʣ˽̤‡ϐȗ͜ƃĤƈϟ\DEVsR˽̤ν‡˜Ł^ƒYl‡ˑͬ\dE bq́ɑDEBVoǶɒ\jMnMōʩʰɍqɥʣ˽̤oPMjDEBVR

™Ô­^ƒmiRNARɛĬ[„Dɢ˂ΧúƟqʾʨ‡ȃĵ\Dɦ̦\jM

ƒYl‡ȶQo\dϨPegtel et al., 2010ϩEydDZhangr¶¬ōʩ

˷˽̤ɕ THP-1 R LPS ~ΠάŅɸ˺nmqƒΩqĶʖoŕǬ\j

miR-150‡ʢʷ˂o EVsĦoƵġ\j˽̤ƒoĭɿ\Db„R͈ˮĦ˄

˽̤ɕHMEC-1q˽̤Ħoŗ‚Ώy„ƒYl‡ȶQo\dEbq́ɑD

ŗ‚ΏˆeEVsßqmiR-150R c-Mybqʾʨ‡ȃĵ^ƒYlkDΞ΂̦

‡Øȵ[`ƒYl‡˔\dϨZhang et al., 2010ϩEƹƁrEVs‡ñ\

dmiRNAqɦ̦‡ͤȶ^ƒd|Dķˤ̱Rˆq˽̤Ǝɯ‡ȃĵ^ƒYl

R^koƊŨ[„jMƒmiR-146aϨLin et al., 2008ϩ‡ʮMjˎˢ‡͉

gdϨKosaka et al., 2010ϩEy_DmiR-146a‡¶¬̣Ġ̫ʰɍ˽̤ɕ

HEK293lŠ¹Ì‘í̜Í̫ʰɍ˽̤ɕCOS-7˽̤Ħkǚʾʨ[`

ƒlDǚʾʨ[`dmiR-146ar˽̤Ħqzn_ƈϟØʋßoPMj}

ơžΰRƎľ^ƒYl‡ȶQo\dEɨMkDCOS-7 o miR-146a ‡ ǚʾʨ[`ƈϟØʋ‡ŵŖ\DŵŖ\dƈϟØʋ‡ķˤ̱Rˆ˽̤ɕ PC3Moʊľ^ƒl˽̤qƎɯȃĵŀɑ‡˔\DmiR-146a qɢ˂ΧúƟ qÕhkLƒROCK1 qʾʨʍƺRͬ|„dE÷Øq́ɑQD˽̤Q

ĭɿ[„dmiRNA rD˽̤ν‡˜Ł\DŘƭĕq˽̤Ħk}ɦ̦^ƒ

YlRͤȶ[„dE[oƹƁr˽̤RmiRNA ‡ĭɿ^ƒɦɠohM j}ŢȹoƊŨ\jMƒEYqȹDĭɿ[„d miRNA r EVs oŦy„

ƒYlRIJȶ\dqkDEVs qĭɿ̀ΆlqξΙ‡ͱvdEEVs qĭɿ

̀ΆorÑ­£ÔÄĦΩƹ̤ǜǻ~˽̤΀ĭ͏ȹoΦij΍ΓD˽̤̲Į ȯ‡͉N͓ŠĀESCRTRξÛ^ƒ̀ΆRˌ„jP‚DYq̀ΆoξÛ

\jMƒAlix¥Ñµ•΀qʾʨ‡ϨHurley et al., 2010ϩDsiRNA‡ʮM

jȃĵ\dE\Q\DmiRNAqĭɿΰoƑŅrnUDYq̀ΆrmiRNA qĭɿoξÛ\jMnMș́ǮR˔[„dEÕȱkDØΒ\d˽̤̲oơ ž^ƒ̧΀qÕhkLƒ¡Ëí‡ʭʬ^ƒΫ˺kLƒDnSMase 2 R EVsqĭɿolMNƊŨ}LƒYlQϨTrajkovic et al., 2008ϩD

siRNA ‡ʮMj nSMase 2 qʾʨ‡ȃĵD}\UrΫ˺ʄǮπƬĹ

GW4869‡ʊľ\dlY…Dĭɿ[„ƒmiRNAqΰRʍƺ\dEydD

˽̤ĦoPWƒʾʨoƑŅr͕„nQgdYlQDmiRNA qĭɿo qzξÛ\jMƒYlR˔ů[„dE\Q\DYqΫ˺qʄǮ‡πƬ\j }ƥĢoĭɿRnUnƒ†WkrnUDóq̀Ά~ΧúƟRξÛ\jMƒ

ș́ǮRL‚DðǢqˎˢRǡd„ƒE

5-5. EVÂԑÔͅˀ΀

EVs o miRNA RŦy„ƒYlRĭQgdqrΑǎkL‚DEVs ʾ͕Ǜ

ırEVs oŦy„ƒͅˀ΀oʁˆRϊygjMdEʢoDEVs oĤΖo Ŧy„ƒEVÂԑÔl͝†„ƒͅˀ΀rDEVsqơž‡ͤȶ^ƒÕhq

ͤȋl\jʮM„jSdE\Q\DŦy„ƒͅˀ΀q˞ϜrƓUϨŹ 5-1ϩD1 ˞Ϝq˽̤Rĭɿ^ƒ EVs Q 100 ˞Ϝ÷Ø}qͅˀ΀RɛĬ [„ƒd|DEVsÂԑÔl͝†„ƒ}qq˞ϜrƓMEydDb„Â ԑÔRmq€Nn˽̤QǤ„d EVs o}Ŧy„ƒQ‡ͪ\U͛ɐ

\dƊŨrnME

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Ź 5-1 EVs o Ŧ y „ ƒ ͅ ˀ ΀

EVs orƓUqͅˀ΀RŦy„Dõ͍˂nͅˀ΀orª¬ËŸµ¯Ñ¹‰ÃÌÔ~

ʞÊ¨•ͅˀ΀DȗˊĭƟDȅŐȚ˔ξΙͅˀ΀nmƓ˞Ɠɡnͅˀ΀RŦy„

jMƒE

ϯ˨6˥ϰExtracellular vesiclesoŦy„ƒͅˀ΀qÂԑÔȖ˻

6-1. ǒͳ

5˥kΒvdRD˽̤Rĭɿ^ƒEVsorɖά~ͅˀ΀RƓUŦy„ƒE

\Q\Dmq€Nnɖά~ͅˀ΀REVsoͧ|Ώy„ƒQrĭɿ^ƒ˽

̤o€gjʷnƒEhy‚DEVsoŦy„ƒɖά~ͅˀ΀‡͛ɐ^ƒYl kDmq€Nn˽̤Rĭɿ\d}qQĭQƒș́Ǯ}LƒlMNYlkL

ƒEydDbqΔkDʷnƒ˽̤˞Qĭɿ[„ƒEVso}ĤΖoŦy

„ƒɖά~ͅˀ΀qơžR˔ů[„ƒEąOsDª¬ËŸµ¯Ñ¹‰ÃÌ ÔqCD63~ CD9DʞÊ¨•ͅˀ΀HSP70Db\jTumor

susceptibility gene 101ϨTSG101ϩnmRDŚUQExosome marker lūs„ƒjMƒE\Q\DɡFn˽̤ʰɍqEVs‡ɵΊ\jDEVso PWƒơžΰ‡ɵΊ^ƒ€NnƊŨrnUDşˎˢ̛Rďijoʷnƒ˽̤

Qĭɿ[„dEVs‡ϊ|jDbqơž‡ˑͬ\d}q‡čϘ\jÂÔ

‘Ôl\jĄʮ\jMƒo^TnMEbYkDɉˎˢkrDRˆ˽̤lϐ Rˆ˽̤‡Ŧ{͓ȫqķˤ̱˽̤ɕlå̱˽̤ɕʰɍqEVs‡ɵΊ\D M†ƒ”EVs Marker”RɉǛomq˽̤ɕʰɍqEVso}Ŧy„ƒqQ ɛ͟\dEbq€NnĭƟ‡Ȗ˻^ƒYlrEVsqơžˑͬ‡Ŧ|DP P€bqŖΰ‡ȂȜ^ƒqoǞˤhl̚O„ƒEydDEVsqǮ΀‡ʢ ǨôWƒ̺MɋȮonƒqkrnMQlɅǡ\Dˎˢoŗ‚˿ˆeE

6-2. ɋȮP€tƪϡȱʀ

6-2-1. Ąʮ\d˽̤ɕlƈϟȱʀ

ķˤ̱˽̤rķˤ̱Rˆ˽̤ɕDPC3DPC-3M-luc ϨPC3MϩD22Rv1b

\jķˤ̱Ø˄˽̤ɕPNT2‡ʮMdEå̱˽̤rDåRˆ˽̤ɕ MDA-MB-231-luc-D3H1 ϨMM231ϩDMDA-MB-231-luc-D3H2LN ϨMM231LNϩDMCF7DMCF7-ADRb\jå̱Ø˄˽̤ɕMCF10A‡ ʮMdEPC3˽̤rķˤ̱RˆŸªÔžIVqǰ̛Qɥˤ[„d˽̤ɕ kϢuqΈ˜Rͬ|„ƒEPC3Mr PC3‡͙ɕl\jD[oΈ˜̦

RϤygd˽̤ɕkLƒE22Rv1˽̤r Prostate Specific Antigen‡ʾ ʨ\DŠÑ­Ï˜ÑΡ»¥ÔυǮkLƒEPNT2˽̤rɩnjķˤ̱Ø˄

˽̤‡SV40o€ƒÚɬŅ[`Dɥˤ[„d˽̤kLƒEMM231˽̤r åRˆǰ̛qΈ˜ĝD̥ɸQɥˤ[„dåRˆ˽̤ɕkL‚D[oÂ

Ÿo˜ɚ\jΈ˜̦‡Ϥ|d˽̤ɕkL‚DŸuq˜ɚk̠~̩u qΈ˜Rͬ|„ƒE[oMM231LNrMM231 ‡͙ɕl\jDΈ˜

̦RϤ|„d˽̤ɕkL‚DŸuq˜ɚoPMjˍɅνkwx

100%qˑˤkΈ˜Rͬ|„ƒEMCF7˽̤}MM231˽̤ŢɡoåR

ˆǰ̛qΈ˜ĝD̥ɸQɥˤ[„dåRˆ˽̤ɕkLƒRŸuq˜

ɚoPMjΘ̮ʼ̦rÿUDŸ¬Ï˜ÑΡ»¥ÔυǮkLƒE

MCF7-ADR˽̤ɕrMCF7 ˽̤‡͙ɕl\jD¥“›Ñ˷ȅRˆĹ­¡

¥“¡ÍoȈȅǮ‡˔^˽̤ɕkLƒEMCF10A ˽̤ɕrå̱Ø˄˽̤

‡κɅν͈ʋʚʊľDϤ‘͝ÄʗǕqƈϟʈkƈϟ\Dɥˤ[„dE MCF10A÷ƒrRPMI 1640 mediumD10%FBSDantibiotic-antimycotic

‡Ŧ{ƈϟʈkƈϟ\DMCF10A ˽̤rMEBMD1% GA-1000D50 μg/ml

hydrocortisoneD1 μg/ml hEGFD500 μg/ml insulinDl 4% BPE ϨLonzaϩ

‡ʊľ\Db„c„q˽̤‡37Ǖ 5%CO2qŒÑ“ȽԥÔĦkƈϟ

\dE

6-2-2. EVsqŵŖȱʀ

EVsrş˞˽̤qƈϟØʋQŵŖ\dEb„c„q˽̤‡15cm«‹¨

Èo80%™Ñ¹ÎѬ˝Ǖkȡ˞^ƒE̘ȳD˽̤Rȗˊ\dYl‡

ˑͬ\jQDFBSÚŦq Advanced RPMI1640 ϨInvitrogenϩo antibiotic-antimycoticlL-–Í¥ÃÑά‡˾ʗǕ2mMonƒ€Noʊ ľ\dƈϟʈoíț^ƒEbqǢD48ȹνƈϟ\DƈϟØʋ‡ŵŖ\D 2,000XgD4Ǖk10ĭνΤǩ\j˽̤ˏʟnm‡ŗ‚σSD[o0.22um q¹‹Í¥Ô‡ʮMj¹‹Í¬ÎԝÊч͉gdE¹‹Í¬ÎԝÊÑ ʌzqƈϟØʋ‡΄ΤǩɦϨOptima XE-90: BECKMAN COULTERϩk 110,000xg÷Øk70ĭΤǩǢDPBS ‡ľOjŢɡqΤǩȤă‡͉MDǤ

„dɽɱʡrEVs‡Ŧ{ʳĭl\jŵŖ\dEŵŖ\dEVsr

Quant-iT™ Protein Assay with Qubit®2.0 Fluorometer ϨInvitrogenϩ

ŸªÄ‡ʮMjͅˀ΀ʗǕ‡ʐƧ\Db„ͅˀ΀ʗǕ‡}lo\j

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6-2-3. ȅĀ

ŽŸ¥ÑºÏ¨¬oʮMdȅĀrÙ͠q͍oyl|dEèɨȅĀr¾Í

“¦Ô¢ɢ͵ȅŸIgG¶©žȅĀϨGE Healthcareϩ}\Ur¾

͐“¦Ô¢ɢ͵ȅ›”IgGÏ´ȅĀϨGE Healthcareϩ‡ʮMdE nPDCD63DCD9DCD81DIntegrin beta 1rϐΨěʣǷkʂŁ\Dɛ Ĭ\dE

6-2-4. Nano particle tracking analysis ϨNTAϩ‡ʮMd˴ƟǠqʐƧ NTAr Nanosight LM10HSlºÍÔÎԜԇʮMjʐƧ\dENTA qʐƧŐʪrʜƷ\dÎԜÔЇ®²˴ƟoǛjjDŕƷЇĴʮ\j ś͗Ņ\j60˚ν·«Ȣǝ\d}q‡˴ƟqºËÑΟŁ‡}lo˴

ƟǠ~ȫ‡£¹¬ŽŠØk͛ɐ^ƒEʐƧorEVs‡PBSkΥƨNjέ

\jʐƧ\dE

6-3. ́ɑ

6-3-1. ŵŖ\dEVsqþˈLjώƟϛǧιo€ƒǜǷ͚ư

ķˤ̱˽̤ɕ4˞Ϝlå̱˽̤ɕ5˞ϜqƈϟØʋQEVs‡΄Τǩʀ ojŵŖ\DbqßQDPC3D22Rv1DPNT2DMM231LNDMCF7 MCF10

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˜²Ä DNA Q mRNA uΈĩDb\jͅˀ΀uq̙͢lMN¡Ñ¬Ë Í­–Âq}lkL‚D[obYQ˽̤D˿̓DŴƦDďĀlnƒo rDɡFn͔ŶRǪ͔kL‚Db„r˜²ÄDzƊeWkrĭQnMEƒ ΩlqˈéăʮDĶʖo€ƒʫƍ˂͔Ŷnmo€gjmq€Nn˜²ÄDz Ɗ‡ĄNQDydbqͱ˰Rή͔kL‚D˽̤Rmq€NnΧúƟ‡ʮM jƒΩĶʖoƳǬ^ƒQrϐnjo̸ŪʉMEɉˎˢkrD˽̤RƒΩqĶ ʖ~ʫƍƑŅoǬˬ\DmiRNA Rmq€NoΧúƟʾʨ‡ǧͱ˰^ƒq QD[orΑǎʁˆ[„jMƒ˽̤νq™Ãȯ—ԝÊÑ©ÔÍkL

ƒ˽̤ƒƹ̤oξ^ƒˎˢ‡͉gdE

˨3˥€‚DmiR-210rÿά˺ʫƍÙDβɧãȹoHIF-1αo€gjʾʨ

ͭƸ[„DŸq¢²–˹¬Æ«Íq͛ɐo€‚Dƪψq̮ʼ˿̓q ÿά˺ʫƍÙkʾʨRØȵ\jMƒYl‡˔\dE\Q\DΖnjά˺Ùk r E2F ¹‰ÃÌÔΧúƟo€gjʾʨͭƸ[„jMƒd|DƥĢoʾʨ

\jMnM†WkrnME€gj miR-210 qʾʨͭƸrʫƍo€gjʷ n‚DũŸqʫƍo€‚ʷnƒΧúƟ‡ɢ˂l\jMƒYl}̚O„ƒE hy‚DŢ] miRNA o}_D˽̤R̔Q„jMƒʫƍo€gjD ʾʨ‡ǧͱ˰^ƒΧúƟ‡ƑOƒYlkD€‚͓ϋnʾʨĵǥ±¨¬ÐÔ

•‡ɠ˲\jMƒqe…NEɉˎˢkrũŸqʫƍo€ƒɢ˂ΧúƟqΦ ȇǮnmrȶQoĬɍnQgdRDðǢqmiRNAˎˢkȶQonƒ

Yl‡Ʌǡ\dME

ɉˎˢkrmiRNARʫƍƑŅo€gjʾʨRƑŁ^ƒÕą‡˔\D˨4

˥kr miR-210rƺnUl} 2 hqβõʹξΙΧúƟqʾʨ‡ˇȗĵǥ

^ƒYl‡ȶQo\dEƺnUl}Dl͠΋\dʪʰr miR-210 qɢ

˂ΧúƟ‡Ģj͛ɐkSd†WkrnMQkLƒEmiRNAqɢ˂Χú Ɵr͓ȫơž\D͓ȫΧúƟqʾʨ‡ĵǥ^ƒYlkÕhqµŸŽŒ‡

ͱ˰^ƒYlrʪoQngjP‚Db„YbʬĀoPWƒmiRNAqĴʙ krnMe…NQEʬʡqΛŅoPMjDͅˀ΀‡™Ô­^ƒΧúƟȫR ϛ̿oƎľ\Dʬʡq͓ϋ[RƎ^DlMN€NnΠ˝rΐnQgdE bqõ†‚oD^koơž^ƒΧúƟqʾʨ‡͓ϋoͱ˰^ƒYlkD͓

ϋ[‡ǜǻ\jSdRDbqÕΩ‡ȊNqRnon-coding RNADl‚†W

miRNAkrnMe…NQEb„r1˥k}Βvd€No˜²Äßqͅˀ

΀‡™Ô­^ƒϗƆqļŠ‡¶¬l̋̈́kɵΊ^„sD¶¬qȱRrƒQ

oļŠrÿUDb„rnon-coding RNARƓUơž^ƒYl‡˔\jMƒE ƪψoƊŨ[„jMƒmiRNAqȫr̋̈́oɵvj¶¬krbqȫR5Đ

÷Øo}nƒEɉˎˢkr˽̤Ħβõʹĵǥoξ\jˈŕ^ƒΧúƟqʾ ʨͱ˰‡ÕhqmiRNAR͉Ną‡˔\dRDˈŕ^ƒăʮ‡ȍhΧúƟ

‡ĵǥ\jMƒd|DmiR-210̵·qʾʨƑŁRʬĀĦk΃Y‚OnMΰ lnƒƋŠDhy‚Dpre-miR-210‡˽̤oƸġ\dψorʁǵRǪ͔k LgdEpre-miR-210 ‡˽̤oƸġ\dψrˇȗɢ˂ΧúƟl\jMƒ

TfR1 qʾʨʍƺR͕„ƒRϨŹ 4-9ϩDY„r ISCU-IRP1̀Άqνȗ

˂nʾʨØȵ€‚}ˇȗ̀ΆqȃĵRǚUęUd|el̚O„ƒEÕȱ kDÿά˺ʫƍÙkmiR-210qʾʨ‡ȃĵ\dƋŠorDTfR1 qʾʨo ƑŅrͬ|„_DʬĀĦk΃Y‚NƒØȵkrTfR1 qʾʨ‡ƑOnM

€Nnò˿zongjMƒqe…NϨŹ4-15ϩEydDpre-miR-210‡˽

̤oƸġ\dψo΃ƒTfR1 qʾʨȃĵq́ɑD˽̤Ǝɯ‡ȃĵ^ƒYl }͕Ĭ\jMƒϨŹ 7-1ϩEYq́ɑR˔^YlrDʫƍƑŅo€ƒ˽̤

qΥǬoɃĴoęUl̚O„ƒmiRNAk^DʬĀĦqΰ‡ʚ͗\d ʾʨØȵrD˽̤oƬ‡Ŕx\Qp_DũŸqʫƍoŠ†`dΥĮnʾʨ ΰqͱ˰Rɺ|„ƒlMNYlkLƒEydDRˆ˽̤rɩnjn˽̤o ɵ v j Ǝ ɯ ^ ƒ d | o TfR1 q ʾ ʨ R Ϥ M Ǫ ͔ R L ‚ Ϩ Ź 7-2ϩD pre-miR-210 o€ƒ TfR1 qʾʨȃĵrɾʽoĴʮkSƒș́ǮRLƒE

\Q\DɖάϨpre-miR-210ϩq«Ì´ÌԝŸªÄq[nƒμʾ~D ɩnj˽̤uqǝϒR˃ʚl}͝OnMd|D̮ʼ˿̓uqʢʷ˂n«Ì´

ÌÔRɺ|„ƒE

˽̤Rƹ̤ϙ˴‡ĭɿ^ƒYlRȶQongjD30 ǎ÷ØR̀Π^ƒ

RDɇ͛ȶnΩĭRƓMEb„rD˽̤RȕĬ^ƒšÃ͎el̚O„d

‚\jDʁˆ[„ƒqRΝQgdYl~DbqƔS[OoŵŖȱʀ~͛

ɐȱʀRρ„jMdYlnmRLƒqkrnMe…NQE\Q\DΑǎ

kr EVs o miRNA RŦy„jMƒʾ͕~ͣȯÂԑÔl\jqɃʮǮ

Rɛͤ[„ƒjMƒnmDʁˆ‡ϊ|ƒˌ͕~çͺRƊŨ[„ƛ|dE[

oȁ͊μʾ}ΛzDNanosightq€NnɦŴ~΄Τǩɦ‡Ąʮ`_EVs

‡ŵŖ^ƒͦ͂nmRʿƋ\DˎˢqǍ‡ȍd`dYlkDEVsR€‚·

Ź 7-1 TfR1 q ʾ ʨ ȃ ĵ o € ƒ

˽ ̤ Ǝ ɯ ̦ u q ǝ ϒ

MCF7˽̤opre-miR-210}\U rTfR1oƳ^ƒsiRNAϨsiTfRϩ

‡Ƹġ\D72ȹνǢo˽̤Ǝɯ̦

‡DMTS‡ʮMdɵ̻Ƨΰʀoj ʐƧ\dE*Pre-miR-NC ydr siNC ‡Ƹġ\d˽̤ƈϟ̖lɵ Ί\jp<0.05E

TfR1

actin

Ź 7-2 R ˆ ˽ ̤ l ɩ nj ˽ ̤ o P W ƒ TfR1q ʾ ʨ ͛ ɐ

ŽŸ¥ÑºÏ¨¬ʀ‡ʮMj MCF7 ˽̤Ϩdžϩl MCF10A

˽̤ϨŝϩoPWƒ TfR1 qʾ ʨ‡ˑͬ\dEÏÔ«‹Ñ–™

ѬÏÔÍractin‡ʮMdE

Αnơžon‚DΑǎq EVs ˎˢqłMrˆ‡͕Ǚƒ}qRLƒEɉˎ ˢkrDbqßk}EVsoŦy„ƒͅˀ΀oˊˆ\DðǢqEVsˎˢo Ǟˤh€NnÂԑÔȖ˻‡͉gdEbq́ɑDķˤ̱˽̤lå̱˽̤ʰ ɍqEVsqɛ́͟ɑQDCD9lCD81rÂԑÔl\jɃʮkLƒś

̦ǮR˔[„dEÕȱkDǣɍÂԑÔl͝†„jMdͅˀ΀k}Dbq ʾʨrʰɍ^ƒ˽̤ɕo€gjˑͬkSnM}q}L‚DEVsqƓɡǮ‡

Ƅν͕ƒYlRkSdEðǢrDƓɡn EVs RʫƍƑŅo€gjĭɿ[

„ƒDbqɠǻǻĭoƑŅRLƒqQnmoˊˆ\dˎˢoŗ‚˿zD˽

̤RʫƍƑŅoǬˬ^ƒψo EVs ‡mq€NoĴʮ^ƒqQȶQo\

dME

÷؇yl|ƒlDΈĩŶƟ~›Œ¬‘ŒÑnmrͱ˰˷‡ÕȱŤoŁQ

^qoƳ\DďFqmiRNA rŢȹo͓ȫqΧúƟʾʨ‡ƓȱŤoĵǥ\

NƒRDɉˎˢkrDmiR-210RϬhqĭƟ‡ɢ˂o\jβõʹ˷kˈŕ

^ ƒ ă ʮ ‡ ʾ ȝ ^ ƒ Y l o € ‚ D ̎ Ư n ĵ ǥ D ^ n † f ǧ ͱ ˰ Ϩmicromanagingϩ‡ȊNYl‡˔\dE\Q} miRNAr•££ÔÄ oĦń[„jD˽̤Ó˿̓ν‡˜Ł^ƒEɉˎˢrDYq€NnȰdn•

˟qͱ˰ŶƟo€ƒȰdnͱ˰qɡǖ‡ͪQo\DʬʡRɠ˲^ƒ͓

ϋnͱ˰˷qò˿z‡ʪ͛^ƒd|qȰˌ͕‡ÛOdE

ϯʹΎϰ

ɉˎˢ‡Μ͉^ƒoLd‚DÖƱnǥȎƸǥϑȣ‡ϓMdȴ˟ʯƔƣƔ ƣςĝΛʪDžƣˎˢ˙qľ̓ƻǫȪȔntoŻˤRˆˎˢ¡Ñ¥Ôˎ ˢǽĭƟ˽̤ɾʽˎˢĭίq̾͹ƢǑĭίκoǩ€‚Ƕʹʱ\ØXy^E ÝĝʬȱorDƓUqYl‡ƣs`jϓSD[oʾ͍~Ƈ˩oξ^ƒƓ Uqɦù‡ϓSy\dE

ƣþͳȭƲɓoPMjD;ήnǥȎƸDǥĿ͝‡Ϳ‚y\dȴ˟ʯƔƣƔ ƣςĝΛʪDžƣˎˢ˙qÞɆ˗ʲȪȔDżɻģɲȪȔoʉUǶʹ̷\y^E

ƹƁƾǺŎƐorDˎˢƫoΪƿ[„jQDŝ}dž}ĭQnM˘‡Ȏ Ƹ\jϓSD[oŻˤRˆˎˢ¡Ñ¥Ôˎˢǽoǽƿ\jQ}ȎƸ‡

̄WjϓSʉUǶʹ̷\y^Eƪϡqǿȁqzn_DˎˢoƳ^ƒ̚O ȱnmƓUqYl‡ƣs`jϓSy\dE

ydDȴ˟ʯƔƣľ̓ˎˢƫqɊʆſÕɶor ǎνkrL‚y\dRD ˎˢƫoΪƿ[„jQƣΩŋɝͳȭykDɡFnƉ˓˂ƪϡǿȁ‡ȪO jϓSy\dEǁɉωñŎƐorͷͳ~Š­´ŒŸ‡Ζ]jȎƸ\jϓS y\dEɉˎˢrƓUqľ̓ˎˢƫqĝΌDǢΌȱqZŌĽRL‚Dʢo ŢɅqzn[yorˎˢqzn_Dˎˢƫʬʄ‡͉gjMUØkƔƑP

Üͨon‚y\dEŢɅqɹʕŭΡɶorƪϡǿʀoξ^ƒɡFnŠ­´

ŒŸ‡ϓSDǶʹ\y^E

ŻˤRˆˎˢ¡Ñ¥ÔˎˢǽĭƟ˽̤ɾʽˎˢĭίq˧ÙȭφŎƐDǃ œʪɥŎƐDêřȼâŎƐ‡r]|l^ƒŢˎˢĭίqzn[yo}ƓU qǥȎƸ‡ϓSDydDˎˢ̛qLƒvSƜ‡͕`jϓSDƔƑĶʖ‡Ř WdYl‡Ƕʹʱ\ØXy^E

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ドキュメント内 Studies on the cellular metabolism controlled by microenvironmental stress (ページ 80-127)