医学毕业论文细胞内钙在人胃癌细胞缺氧诱导因子1表达及转录激活中的作用

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1、细胞内钙在人胃癌细胞缺氧诱导因子1表达及转录激活中的作用姓名：2014年6月25日细胞内钙在人胃癌细胞缺氧诱导因子1表达及转录激活中的作用作者：兰梅，时永全，韩者艺，宁晓喧，樊代明【摘要】目的观察细胞内钙变化对氧调节性亚单位HIFla表达及其HIF1转录激活的影响。方法常氧吋，采用通透性细胞内钙的螯合剂BAPTAAM或钟的离子载体Ionomycin降低或升高细胞内钓，用Westernblot检测其对SGC7901细胞巾HIFla蛋白表达的影响，然后采用间接免疫荧光法、双荧光素酶报告系统及ELISA法检测改变细胞内钙对HIFla转位、HIF1转录活性及其革巴基因血管

2、A)皮也长

3、因子(Vascularendothelialgrowthfactor，VEGF)分泌的影响。结果用BAPTAAM降低细胞内钙，能诱导HIFla的表达，促进HIFla的转位，增强HIF1的转录活性，增加HIF1调节基因VEGF的分泌。用Ionomycin升高细胞内轉，虽然也有微弱的促HIFla稳定及转位作用，但对HIF1转录活性及VEGF的分泌并无明显影响。结论螯合细胞内钙能诱导胃癌细胞屮HIFla的表达及HIF1的转录激活，提示细胞内钙变化可能在H癌细胞的缺氧信号转导过程中发挥了重要作用。【关键词】胃癌细胞；细胞内钙；缺氧诱导因子1；蛋白表达；转录活性Departmentof

4、InternalMedicine,theAffiliatedHospitalofMedicalCollegeofShaoxingUniversity,Shaoxing312000,China;InstituteofDigestiveDiseaseStateKeyLaboratoryofCancerBiology，XijingHospital,FourthMilitaryMedicalUniversityAbstract：ObjectiveTostudythepossiblerolesofchangedintracellularcalciumlevelinSGC7901cell

5、sonHIFlaexpressionandHIF1transcriptionalactivity.MethodsUndernormoxicconditions，theproteinexpressionofHIFlainSGC7901cellsaftertreatmentwithBAPTAAM(acellpermeantCa2+chelator),orionomycin(ancalciumionophore)wasobservedbyWesternblot.TheeffectofBAPTAorionomycinonnuclearlocationofHIFlot，HIF1luci

6、feraseactivityaswellassecretionofvascularendothelialgrowthfactor(VEGF)，atypicalHIF1targetgene，wasdeterminedbyindirectimmunofluorescenceassay,dualluciferasereportersystemandenzymelinkedimmunosorbentassay(ELISA)respectively.ResultsDecreasingtheintracellularcalciumlevelbyBAPTAinducedHIFlotnucl

7、earaccumulation,stimulatedHIF1dependenttranscriptionandenhancedexpressionofVEGFinSGC7901cellsundernormoxicconditions.However，theinductionofHIF1activityandthesecretionofVEGFwerenotcausedbyionomycinapplication，thoughitcanmediatedaweakandtransientHIFlotnuclearaccumulation.ConclusionChelationof

8、cellularcalciummodulatesHIFlaexpressionandHIF1transcriptionalactivityinhumangastriccancercells.Changesinintracellularfreecalciumconcentrationappearedtohaveacriticalroleinhypoxicsignaltransductioningastriccancercells.Keywords：Gastriccancercells;Intracellu