靶向端粒酶催化亚单位基因hEST2反义寡核苷酸对人肝癌细胞生长的抑制作用.doc

《靶向端粒酶催化亚单位基因hEST2反义寡核苷酸对人肝癌细胞生长的抑制作用.doc》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、靶向端粒酶催化亚单位基因hEST2反义寡核苷酸对人肝癌细胞生长的抑制作用作者：杨波脱朝伟张宁刘秋珍王明耀【摘要】目的观察靶向端粒酶催化亚单位基因hEST2反义寡核苷酸对人肝癌细胞生长的抑制作用。方法从Genebank中钓取人端粒酶催化亚单位基因hEST2的mRNA序列，通过计算机模建二级结构辅助设计并合成硫化修饰反义寡核苷酸(癌泰得)。采用人肝癌裸小鼠原位移植模型(HCMY89)。实验分为生理盐水组，5FU10mg/kg组，癌泰得50mg/kg组、75mg/kg组，癌泰得75mg/kg联合5FU10mg/kg组、50mg/kg联合5FU10m

2、g/kg组。尾静脉给药，每天1次，连续20d。观察移植瘤质量和抑瘤率、移植瘤体积、AFP浓度和移植瘤组织学。结果(1)5FU10mg/kg组的抑瘤率为27.85%，其他各治疗组的抑瘤率均在42.14%～74.29%之间；(2)各治疗组瘤质量、瘤体积和AFP浓度均低于生理盐水组且差异有统计学意义；(3)瘤体积和AFP浓度之间呈直线关系，为显著正相关(r=0.9977)；(4)癌泰得和5FU治疗人肝细胞癌后，肝癌细胞出现不同程度的凋亡、坏死，同时伴有纤维组织增生。结论癌泰得对肝癌细胞生长具有抑制作用，疗效优于5FU；与5FU合用具有协同抗肿瘤效应

3、。【关键词】端粒酶催化亚单位；反义寡核苷酸；原发性肝细胞癌；原位移植；裸小鼠；疾病模型　　InhibitoryeffectofantisenseoligodeoxynucleotidetargetedagainsttelomerasecatalyticsubunitgenehEST2onthegrowthofhumanhepatocellularcarcinomacells【Abstract】ObjectiveToinvestigatetheinhibitoryeffectofantisenseoligodeoxynucleotidetargeted

4、againsttelomerasecatalyticsubunitgenehEST2onthegrowthofhepatocellularcarcinoma(HCC)cells.MethodsmRNAsequenceofhEST2wasobtainedfromGenebankdatabase.ComputerwasappliedinhelpingimitatethesecondarystructureofmRNAanddesignantisenseoligodeoxynucleotide.Chemosynthesismethodwasusedtosy

5、nthesizethesulfurizedantisenseoligodeoxynucleotidetargetedagainsthEST2(Cantide).Orthotopicimplantationmodelofhepatocellularcarcinomainnudemice(HCMY89)wasestablished.Allmiceweredividedintonormalsalinegroup(negativecontrol),5FU10mg/kggroup(positivecontrol),Cantide50mg/kggroup,C

6、antide75mg/kggroup,Cantide75mg/kgplus5FU10mg/kggroup,Cantide50mg/kgplus5FU10mg/kggroup.Venacaudalisinjectionwasdoneonallmiceonceperdayfor20dayscontinuouslytoobserveweight,volume,histologyandinhibitionrateofimplantedtumors,AFPconcentration.Results(1)5Inhibitionrateoftumorwas27

7、.85%in5FU10mg/kggroupbut42.14%74.29%inothergroups.(2)Inthetreatmentgroup,thetumorhadlighterweight,smallervolumeandlowerAFPconcentrationthanthoseinnormalsalinegroup.(3)AFPconcentrationhadapositivecorrelationwithtumorvolume(r=0.9977).(4)Apoptosisandnecrosisofcancercellsandproli

8、ferationoffibroustissuewereobservedaftertreatingHCCby5