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《mir--183靶向抑制pdcd4促進食管鱗癌細(xì)胞增殖及侵襲》由會員上傳分享，免費在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、南京醫(yī)科大學(xué)碩士學(xué)位論文mechanismsinESCCdevelopment．Methods：1．UsingmicroarrayanalysistoscreenaberrantlyexpressedmiRNAsin3pairsofESCCtissuesandnormalcontrols，mil0183wasselectedastheobjectiveofthisstudy．2．TheexpressionofmiR-183in32pairsofESCCtissuesand30pairsofesophagealintraepithelialneop
2、lasiatissueswereexaminedbyTaqrmnqRT-PCRwhilepdcd4(mRNAandprotein)wasdetectedbySYBRGreenqRT-PCR,WesternblotandIHCassay．3．ThetargetgemsofmiR-183weresearchedwithbioinformaticstoolssuchasTargetScan／miRBaseandPictar,potentialtargetgenesexpressionwereassessedbyqRT-PCI乙Westernblotan
3、dLuciferasereporterassay．4．MiR-183mimics，miR-183inhibitors，si-pdcd4andtheircorrespondingnegativecontrolsweretransfectedintoESCCcells，CCK-8assayforcellprolifemtiort,colonyformationassayforcbnogenicgrowth,flowcytometryassayforcellapoptosis，transwellassayforcellmigrationandinvas
4、ion．5．DifferentialconcentrationsofP13眥TinhibitorLY294002(0rtM、10州、200M、40山VI)wereaddedintoESCCcellsandculturedforhours，miR-183levelWasmeasuredbyqRT-PCR,pdcd4，AKT,P-AKTexpressionsweredetectedbyWesternblotassay．Results：1．51up-regulatedand17down-regulatedmiRNAsinourmicmarray,wit
5、hmiR-183levelinESCCwas4．24timesthannormalcontrols．TheexpressionofmiR-183in32pairedofESCCtissuesWas3．98timesthannormalcontrols儼<0．0001)，while2．93timesinintraepithelialmophsiatissuesfP<0．005)．Theexpressionofpdcd4mRNAandproteininESCCtissueswereO．46and0．27timesthannormaltissuesfP
6、<0．05)．．5．南京醫(yī)科大學(xué)碩士學(xué)位論文AccordingtotheIHCassay,thestainingofpdcd4proteininESCCtissueswasf0．178±0．021)，whichwassignificantlylowerthanthatofnormaltissues(0．267±0．031)伊<0．05)．2．Pdcd4wasonetargetgeneofmaR-183accordingtobioinforrmticsanalysis．Theexpressionofpdcd4proteinwasreduced(in
7、creased)whentmnsfectedwithmiR-183mimics(inhibitors)儼<0．05)whilepdcd4mRNAdidn’tchange(JP>0．05)．LuciferaseactivityWasdecreasedwhentransfectedpdcd4wt-3'-UTRvectorinmiR-183mimicscomparedtocontrol伊8、ffectsofmiR-183mimicsonESCCcellsweredescribedusingCCK-8，colonyformat
