LIM蛋白質(zhì)Ajuba增強(qiáng)乳腺癌細(xì)胞ERɑ介導(dǎo)的轉(zhuǎn)錄調(diào)控.pdf

1、上海交通大學(xué)醫(yī)學(xué)院碩士學(xué)位論文上海交通大學(xué)醫(yī)學(xué)院碩士學(xué)位論文LIM蛋白質(zhì)蛋白質(zhì)Ajuba增強(qiáng)乳腺癌細(xì)胞增強(qiáng)乳腺癌細(xì)胞ERɑ介導(dǎo)的轉(zhuǎn)錄調(diào)控介導(dǎo)的轉(zhuǎn)錄調(diào)控LIMproteinAjubaactivatesERɑmediatedtranionalregulationofbreastcancercells研究生姓名：徐北惠學(xué)號(hào)：研究生姓名：徐北惠學(xué)號(hào)：1137209155申請(qǐng)學(xué)位級(jí)別：醫(yī)學(xué)碩士指導(dǎo)教師：倪培華副教授培養(yǎng)單位：上海交大醫(yī)學(xué)院附屬瑞金

2、醫(yī)院學(xué)科專業(yè)：臨床檢驗(yàn)診斷學(xué)研究方向：腫瘤發(fā)生發(fā)展的機(jī)制研究基金資助：上海市科委基礎(chǔ)研究領(lǐng)域項(xiàng)目（申請(qǐng)學(xué)位級(jí)別：醫(yī)學(xué)碩士指導(dǎo)教師：倪培華副教授培養(yǎng)單位：上海交大醫(yī)學(xué)院附屬瑞金醫(yī)院學(xué)科專業(yè)：臨床檢驗(yàn)診斷學(xué)研究方向：腫瘤發(fā)生發(fā)展的機(jī)制研究基金資助：上海市科委基礎(chǔ)研究領(lǐng)域項(xiàng)目（81402177）論文提交日期：）論文提交日期：2016年4月答辯日期：月答辯日期：2016年4月LIMproteinAjubaactivatesERɑmediated

3、tranionalregulationofbreastcancercellsABSTRACTObjective：ToexpletheroleofLIMproteinAjubainthetranionregulationofestrogenreceptɑ(ERɑ)signalingpathway.Methods:TheinteractionbetweenAjubaERɑwasdemonstratedbyendogenousaswellas

4、exogenousimmunoprecipitationtechiniquesGSTpulldownassay.RNAhighthroughputsequencinganalysiswasperfmedinT47DcelllineswithstableknockdownoverexpressionofAjubasoastofindoutERɑdownstreamtargetgenesregulatedbybothestrogenAjub

5、a.ThefindingwasvalidatedbyRTqPCR.LuciferaserepterassaywasusedtoexaminetheactivationofAjubaonthepromotofTFF1onedownstreamtargetgeneofERɑfindouttheexactAjubabindingsite.CellcountingKit8assay(CCK8)wasperfmedtoidentifythecom

6、binedeffectofAjubaestrogenoncellgrowthactivityinT47DcellswithstableoverexpressionknockdownAjubaMCF7cellswithstableoverexpressionofAjubarespectively.ThesensitivitytotamoxifenwasalsodeterminedintheaboveT47DMCF7celllines.Re

7、sults:DirectcombinationofAjubawithERɑindependentlyofestrogencanbefoundbutthiscombinationcanbefurtherenhancedbytheinfluenceofestrogen.AseriesofERɑtargetgenesregulatedbyAjubawerescreenedoutbyRNAseqthefindingswerereliablyva

8、lidatedbytheRTqPCRtechnique.ThecomplexAjubaERɑactivatedthetranionalactivityofERɑdownstreamtargetgeneTFF1bybindingtheestrogenresponsibleelement(ERE)onitspromot.AjubaERɑhaveasynergisticpromotingeffectonthegrowthofERpositiv

9、ebreastcancercells.AjubadeclinedthesensitivitytotamoxifeninERpositivebreastcancercelllines.Conclusion:AjubaanauxiliaryactivatingproteincanpositivelyregulatethesignalingpathwaymediatedbyERɑ.Keywds:Ajubaestrogenreceptɑbrea