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《臨床醫(yī)學(xué)畢業(yè)論文衰老細(xì)胞中調(diào)控胰島素樣生長(zhǎng)因子結(jié)合蛋白3的表達(dá)》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫。
1、衰老細(xì)胞中調(diào)控胰島素樣生長(zhǎng)因子結(jié)合蛋白3的表達(dá)姓名，2014年6月25日作者：任文君，王群義，呂小風(fēng)，毛澤斌，蔣曉剛【摘要】目的：研究在衰老細(xì)胞中調(diào)控胰島索樣生長(zhǎng)因子結(jié)合蛋白3(IGFBP3)表達(dá)增加的影響因素。方法：用Northernblot的方法顯示IGFBP3基因的表達(dá)在年輕和衰老的2BS細(xì)胞巾存在差異;PCR擴(kuò)增出人類IGFBP3上游包括5'UTR區(qū)的2kb的序列并用酶切得到4組不同長(zhǎng)短的IGFBP3啟動(dòng)了?片段；將各片段用EffecteneTransfectionReagent試劑盒轉(zhuǎn)染到年輕和衰老細(xì)
2、胞中，測(cè)出幾組構(gòu)建基兇片段的啟動(dòng)活性，確定町調(diào)控轉(zhuǎn)錄活性的區(qū)域；通過重疊寡核苷酸凝膠阻滯實(shí)驗(yàn)確定在該活性區(qū)域屮的增強(qiáng)子元件。結(jié)果：與年輕的2BS細(xì)胞和比，衰老的2BS細(xì)胞巾IGFBP3基因的表達(dá)升高；在IGFBP3啟動(dòng)子+59到-58序列之間存在著蛋白結(jié)合位點(diǎn)；5'ccagcctgccaagcagcgtgccccggttgc3'是IGFBP3的培強(qiáng)了元件。結(jié)論：在衰老的2BS細(xì)胞中IGFBP3基兇上游-37到-8的30bp序列存在一個(gè)新的増強(qiáng)子元件IEE(IGFBP3enhancerelement),對(duì)IGFB
3、P3的表達(dá)起到調(diào)控作用。【失鍵同】胰島素樣生長(zhǎng)因子結(jié)合蛋白3;IEE(IGFBP3enhancerelement);2BS細(xì)胞[Abstract]AIM:Tostudytheinfluencingfactorsoftheincreasedexpressionofthecontrollinginsulingrowthfactorsbindingprotein3(IGFBP3)inthesenescentcells.METHODS:Northernblotwasusedtoshowofthedifferential
4、expressionoftheIGFBP3geneintheyoungandsenescent2BScells;Thesizeof2kbhumanIGFBP3upstreamsequenceincludingtheseriesofthe5'UTRareawasamplifiedbyPCR,andfourgroupsofIGFBP3promoterfragmentsofdifferentlengthswereobtainedbyenzymedigestion.TheEffcctcncTransfectionRea
5、gent(Qiagen)kitwasusedtotransfectthefragmentsintotheyoungandsenescentcells.Thepromotingactivityofseveralgroupsofconstructinggenefragmentswereevaluated.Theareaofthecontrollingtransfectionactivitywasdetermined.Theenhancerelementintheactivityareawasascertainedb
6、ysuperimposingtheoligonucleotidegelblockingexperiment.RESULTS:Comparedwiththeyoung2BScells,theexpressionoftheIGFBP3geneinthesenescent2BScellswasenhanced.Therewasaproteinbindinginthefragmentsitefromsite+59to58oftheIGFBP3enhancer.5'ccagcctgccaagcagcgtgccccggtt
7、gc3ZwastheenhancerelementofIGFBP3.CONCLUSION:Inthe30bpfragmentfromsite37to8oftheIGFBP3geneupstream,thereisanewIGFBP3enhancerelementIEE,whichplaysacontrollingroleintheexpressionofIGFBP3.［Keywords］Insulingrowthfactorbindinprotein3;IEE(IGFBP3enhancerelement);2B
8、Scells胰島素樣生K:因子(insulingrowthfactors,IGFs)是人體最重要的生長(zhǎng)因子之一，對(duì)人體組織細(xì)胞的生長(zhǎng)發(fā)育有廣泛而重耍的調(diào)節(jié)作用。不少研究結(jié)果發(fā)現(xiàn)胰島素樣生長(zhǎng)因子1(IGFI)對(duì)免疫系統(tǒng)及多種免疫細(xì)胞宥正性調(diào)節(jié)作用，如促進(jìn)T、B淋巴細(xì)胞和粒細(xì)胞增生［1］，調(diào)節(jié)前B細(xì)胞分化，促進(jìn)B細(xì)胞抗體生成［2］，增強(qiáng)NK細(xì)胞殺傷活性等［3］。許多免疫組織細(xì)胞也表達(dá)IGF
