TSA联合基因工程腺病毒H101治疗人食管癌裸鼠皮下移植瘤的研究冯婷1刘霞'董子明彳1#通讯作者：男54岁教授博士研究生导师研究方向：病理与病理生理学Email：dongziming@zzu.cdu.cn(1郑州大学基础医学院病理生理教研室河南郑州450052)摘要目的：研究TSA药物对基因工程腺病毒H101杀伤食筲癌EC9706细胞作用的影响，探讨HDAC抑制剂在腺病毒载体基因治疗中应用的可能性和作用机制。方法：①先成瘤后治疗组，构建裸鼠食管癌移植瘤模型，待肿瘤长出1W后，分组：TSA治疗组(瘤内注射1.Oymol/LTSA/只欣)，H101治疗组(瘤内注射50X108vp/d),TSA联合H101治疗组即先注射TSA,第二天注射H101,注射剂量同上。另设空白对照组，以上治疗持续两周，治疗结束取瘤组织检测。②先处理后成瘤组分别采用浓度为4.0piol/LTSA体外处理EC906细胞48h,浓度200X108vp/dH101处理EC9706细胞48h,及二者联合处理EC9706细胞48h,同样设置空白对照组，将处理后的细胞种植于裸鼠皮下构建裸鼠食管癌移植瘤模型。待瘤体长出2周后，取瘤组织用RealTimePCR.Western-bioting和免疫组化法检测CAR在移植瘤中的表达，测量肿瘤体积。结果：无论是先处理后成瘤组还是先成瘤后治疗组TSA组和TSA联合H101治疗组均可上调裸鼠食管癌移植瘤组织中CAR蛋白的表达，相比与对照组和H101治疗组均有明显差界(/K0.05),但TSA组和TSA联合H101治疗组之间相比差异无显著性(Q0.05)。而对人食管癌EC9706细胞裸鼠皮下移植瘤大小影响TSA联合H101治疗组与TSA组、H101单独治疗组相比均冇明显差异(/K0.05),TSA和H101组相比差异无显著性(00.05)。各治疗组与空白对照组相比，差异均冇统计学意义(ZK0.05)。结论：TSA能増强H101治疗人食管拘裸鼠皮下移植瘤，其作用机制是TSA±调了食管癌CAR蛋白的表达。关键词：柯萨奇-腺病毒受体(Coxsackieandtidonov让usreceptor,CAR):TrichostatinA(TSA)；基因工程腺病毒；EC9706；裸鼠［中图分类号］:R735.1；R730.231［文献标识码］:ATSAcombinedgeneticallyengineeredadenovimsH101fortreatmentoftumortissueofnudemicetransplantedwithhumanesophagealcancercellsfengting1liuxia1dongziming1^DepartmentofPathophysiology,CollegeofBasicMedicalSciences,ZhengzhouUniversity,Zhengzhou450052,China[ABSTRACT]Objective：thestudyistoobserveTSAdrugfortheeffectofgeneticengineeringadenovirusH101killingesophagealcancerEC9706cell9anddiscusstheactionmechanismsandthepossibleapplicationsofHDACinhibitorinadenovirusvectorsforgenetherapy•Methods:①thefirstexaminationistreatmentaftergrowingtumour•theexaminationfollowingisbuildingnudemodeloftransplantedtumorinesophagealcancer,andgroupingwhenthetumorgrowingoneweek.TSAtreatmentgroup(injectionTSAl.Opmol/LTSA/only/time);H101treatmentgroup(injectionH10150x108vp/d);TSAandH101treatmentgroup,thefisttoinjectTSA,andthenextdaytoinjectHlOlJheinjectiondosageasabove,anothergroupismatchedgroup.Obtainingtumortissueaftertreatingtwoweeks.②thesectionexaminationistreatmentbeforegrowingtumour.onegroupisutilizingtheconcentrationof4.0pmol/LTSAtotreatEC906cells48h,tnesecondgroupisutilizingtheconcentrationof200x108vp/dHIOltotreatEC9706cells,48h,tnethirdgroupisthetwodrugsinjoinedtotreatEC9706cells48h,anothergroupismatchedgroup•thetreatedcellsgrownundernskinofnudemicetobuiltxenograftmodelofesophagealcancer•Aftertwoweeks^obtainingthetumortissuesandusingRealTimePCR,WesterbiotingandimmunohistochemistrytodetectexpressionofCARinthetumorandmeasuretumorvolume.Results:.Whetherthefirstexaminationoftumororthesectionexamination9compareingthematchedgroupandonlyH101treatmentgroup，TSAtreatmentgroupandTSAassociatingwithH101treatmentgroupabletorisetheexpressionofCARproteininesophagealcancexenografttissueinnudemice(P<0.05).ButtheTSAgroupandtheTSAcombinedHl01groupwerenosignificantdiffe...