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《sirna沉默traf6基因?qū)κ彻馨〆c109細胞增殖及凋亡的影響》由會員上傳分享，免費在線閱讀，更多相關(guān)內(nèi)容在學術(shù)論文-天天文庫。
1、汕頭大學醫(yī)學院碩士學位論文TRAF6siRNAandNegativeControlsiRNA;2.24haftertransfection,cellsweredigestedbyRNAisoReagenttoextracttotalRNA;RT-PCRwascarriedouttoconfirmthetransfectioneffect;3.Aftertransfectionfor24h,adherentcellswerecollected,washedwithPBS,centrifugedandthensolublesubcellularpro
2、teinswereextractedbyproteinextractionkit.WestenBlotwasusedtoexploitTRAF6andP65proteinsexpressionineachgroups.4.TransfectedbysiRNAfor24h,adherentcellswerecollected,washedwithPBS,centrifuged,fixedwith4%polylysine,andthendealwith0.5%tritonX100.Afterantibodyincubationfluorescence
3、intensityofTRAF6wasdetectedbyflowcytometry;5.TransfectedbysiRNAfor24h,thecellproliferationwasmeasuredbyMTTassay,andapoptosiswastestedbyflowcytometry.6.ExperimentaldatadealedwithSPSS13.0statisticalanalysissoftware.Results:1.AsiRNAwhichcouldeffectivelyblockTRAF6geneexpressionwa
4、sidentifyed,andtheoptimumconcentrationwasfound.2.ItshowseffectivelyinterferenceofTRAF6genebyRT-PCR,WesternBlotandflowcytometry.3.AfterTRAF6genewasblocked,itshowsthatP65proteinexpressedlowerinnuclearextractbywesternblot;4.ThecellularmorphologyofEC109cellshasnosignificantchange
5、betweencellstreatedwithTRAF6siRNAandcellswithNegativeControlsiRNA.TRAF6genedoesn’taffecttheproliferationofEC109cells,butcanaffectapoptosisofEC109cells.Conclusions:TRAF6genemayplayanimportantroleinindevelopmentandprogressionofhumanesophagealcancer,andthemechanismmayinduceexces
6、siveactivationofNF-κBinEC109cellstocarryouttheinhibitingapoptosisfunctions.Keywords:Apoptosis,RNAi,SignalTransduction,TRAF6,NF-κBIV汕頭大學醫(yī)學院碩士學位論文縮略詞表縮略詞英文中文AcrAcrylamide丙烯酰胺APAmmoniumpersulfate過硫酸銨BSAbovineserumalbumin牛血清白蛋白bpBasepair堿基對cDNAComplementarydeoxyribonucleicacid互補脫
7、氧核糖核酸DEPCdiethypyrocarbonate焦碳酸二乙酯DMSODimethylSulfoxide二甲基亞砜DMEMDulbecco’smodifiedeagle’smedium達爾伯克改良伊格爾培養(yǎng)基dNTPDeoxynucleotidetriphosphate脫氧核苷三磷酸FBSfatalbovineserun胎牛血清TRAFstumornecrosisfactorreceptorassociatedfactor腫瘤壞死因子受體相關(guān)因子TNFRTumornecrosisfactorreceptor腫瘤壞死因子受體TRANCETNF
8、-relatedActivation-InducedCytokine腫瘤壞死因子相關(guān)激活誘導因子NF-κBnucleartranscri
