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《臨床醫(yī)學(xué)畢業(yè)論文衰老細胞中調(diào)控胰島素樣生長因子結(jié)合蛋白3的表達》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、衰老細胞中調(diào)控胰島素樣生長因子結(jié)合蛋白3的表達姓名,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:研究在衰老細胞中調(diào)控胰島索樣生長因子結(jié)合蛋白3(IGFBP3)表達增加的影響因素。方法:用Northernblot的方法顯示IGFBP3基因的表達在年輕和衰老的2BS細胞巾存在差異;PCR擴增出人類IGFBP3上游包括5'UTR區(qū)的2kb的序列并用酶切得到4組不同長短的IGFBP3啟動了?片段;將各片段用EffecteneTransfectionReagent試劑盒轉(zhuǎn)染到年輕和衰老細
2、胞中,測出幾組構(gòu)建基兇片段的啟動活性,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驗確定在該活性區(qū)域屮的增強子元件。結(jié)果:與年輕的2BS細胞和比,衰老的2BS細胞巾IGFBP3基因的表達升高;在IGFBP3啟動子+59到-58序列之間存在著蛋白結(jié)合位點;5'ccagcctgccaagcagcgtgccccggttgc3'是IGFBP3的培強了元件。結(jié)論:在衰老的2BS細胞中IGFBP3基兇上游-37到-8的30bp序列存在一個新的増強子元件IEE(IGFBP3enhancerelement),對IGFB
3、P3的表達起到調(diào)控作用?!臼фI同】胰島素樣生長因子結(jié)合蛋白3;IEE(IGFBP3enhancerelement);2BS細胞[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因子之一,對人體組織細胞的生長發(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)胰島素樣生長因子1(IGFI)對免疫系統(tǒng)及多種免疫細胞宥正性調(diào)節(jié)作用,如促進T、B淋巴細胞和粒細胞增生[1],調(diào)節(jié)前B細胞分化,促進B細胞抗體生成[2],增強NK細胞殺傷活性等[3]。許多免疫組織細胞也表達IGF