ha纳米载体转hgm-csf hepg2疫苗联合多柔比星抗肝癌实验分析word论文.docxVIP

HA纳米载体转hGM-CSFHepG2疫苗联合多柔比星抗肝癌实验研究研究生：赵祺导师：段晓明教授摘要目的：评价HA纳米载体转hGM-CSF基因的HepG2疫苗联合多柔比星治疗肝癌的效果。方法：建立Hu-PBL-SCID HepG2 荷瘤小鼠模型，待皮下种植瘤长至100-150 mm3 时，随机分为5 组，每组5 只：(1)对照组(PBS)；(2)化疗组(多柔比星DOX)；(3) 疫苗组（60Co照射的转染GM-CSF基因的HepG2细胞）；(4)先疫苗后化疗组；(5)先化疗后疫苗组。末次治疗后第5 天处死各组小鼠，用MTT 法测定脾脏CTL 活性及肿瘤组织浸润淋巴细胞(TIL)杀伤活性；取肿瘤组织行HE染色及CD8+T 细胞免疫组化检查。结果：化疗组的CTL活性与对照组相比无显著变化（P0.05）。疫苗组和先疫苗后化疗组及先化疗后疫苗组的CTL活性显著强于上述两组（P0.05），该三组两两之间也存在显著差异（P0.05），而先化疗后疫苗组的CTL活性增强最为明显，明显高于其他两组（P0.05）。该组的TIL活性同样强于其他各组。同其它组相比，先化疗后疫苗组的瘤体内可见大量肿瘤细胞坏死，CD8+T 细胞浸润显著增加。结论:HA纳米载体转染hGM-CSFHepG2疫苗的主动免疫联合多柔比星化疗具有协同抗肝癌作用，为肿瘤免疫联合化疗治疗肿瘤提供了实验依据。关键词：肝癌；肿瘤疫苗；多柔比星；GM-CSF基因；免疫化疗3Anti-hepatomaEffectsofHepG2TumorVaccineTransfectedhGM-CSFGeneMediatedbyHANanoparticlesCombinedwithDoxorubicinABSTRACTObjective:ToinvestigatethetreatmenteffectsofHepG2tumorvaccinetransfectedhGM-CSF genemediatedbyHAnanoparticlescombinedwithdoxorubicintolivercancer.Methods:EstablishHu-PBL-SCIDHepG2tumor-bearingmice.Whenthe xenograftedto100-150mm3,micewererandomlydividedintofivegroups,fiverats ineachgroup:(1)controlgroup(PBS);(2)chemotherapygroup(doxorubicinDOX);thevaccinegroup(HepG2cellstransfectedwithGM-CSFgeneafter60Co irradiated);(4)chemotherapyafterthevaccinegroup;(5)thevaccineafter chemotherapygroup.Themiceweresacrificed5dafterthefinaltherapyand examinedtheirspleensCTLactivityandtumorinfiltratinglymphocyte(TIL)activity measuredbyMTT;tumortissuewasstainedbyHEandCD8Tcellsinthetissuewas examined byimmunohistochemistry.Results:TheCTLactivityofchemotherapygrouphadnosignificantchangewhich comparedwithcontrolgroup(P0.05).theCTLactivityofThevaccinegroupand chemotherapyaftervaccinegroupandthevaccineafterchemotherapygroupwere significantlystrongerthantheabovetwogroups(P0.05).Thereweresignificant differencesbetweenthethreegroup(P0.05),whiletheCTL activityofthevaccine afterchemotherapygroupincreasedmostsignificantly,higherthantheothergroups (P0.05).TILactivityofthisgrouphasbettereffectthanothers.Comparedwith other groups, more tumor necrosis in tumor specimens and more inf