细胞因子light和ifn-γ联合转染hepg2细胞及对其凋亡影响的实验研究-外科学(肝胆外科)专业论文.docx

细胞因子LIGHT和IFN一丫联合转染HepG2细胞及对其凋亡影响的实验研究中文摘要目的：观察脂质体介导的LIGHT和IFN吖配比转染HepG2细胞后对其凋亡及Fas和FasL表达的影响。方法：用本实验室已成功制备的LIGHT和IFN-y的质粒DNA转化E．coli JM一109感受态大肠杆菌，提取质粒；将HepG2细胞分为LIGHT基因单独转染组、LIGHT和IFN吖基因联合转染组及空白对照组，处理后分别于12h，24h，48h收集HepG2细胞，用流式细胞仪检测HepG2细胞凋亡及Fas和FasL的表达情况。结果：转染后，Fas和FasL在HepG2细胞高表达，但FasL不如Fas升高程度显著。Fas在LIGHT基因单独转染组表达率高于空白对照组，而LIGHT和IFN吖联合转染组又高于LIGHT基因单独转染组(P0．01)；LIGHT和IFN—Y联合转染组在12h和24h FasL的表达明显高于LIGHT基因单独转染组(P0．01)，在48h低于LIGHT基因单独转染组(尸0．01)。转染后HepG2细胞随时间的延长凋亡率增高，且LIGHT基因单独转染组细胞凋亡率高于空白对照组，而LIGHT基因和IFN-y 联合转染又高于LIGHT基因单独转染组(尸O．01)。结论：利用脂质体成功将LIGHT和IFN吖的质粒DNA转染HepG2细胞，转染后HepG2细胞Fas和FasL表达上调，细胞凋亡率增高，且LIGHT基凶和IFN叫基因联合转染效果优于LIGHT基因单独转染。硕士研究生王正华(肝胆外科)指导教师吴力群教授关键词：LIGHT；IFN-q,；Fas／Fas—L；转染；细胞凋亡；肝癌THESTUDYOFAPOPTOSISINHEPG2CELLSAFTERCOMBINEDTRANSFECTIONWITHLIGHTANDIFN一丫Abstractobjective：ToinvestigatetheexpressionofFas、FasLandtheapoptosisoflivercancercelllineHepG2transfectedwithLIGHTandIFN-YgenemediatedbyCationicliposome．Methods：DividingHepG2cellsintothesolotransfectionofLIGHTgene、thecombinedtransfectionofLIGHTandIFN-Y genesandthecontroll，thentransfectHepG2cellsmediatedbyCationicliposomewithpcDNA4C．LIGHTcDNAandpcDNA4／HisMax—hlFN．YcDNAextracted fromE．coliJM．109andcellectingHepG2cellsonl2h，24hand48haftertransfection．toinvestigatetheapoptosisofHepG2cellsandtheexpressionofFasandFasLofHepG2cellswithflowcytometry．Results：Atiertransfection，theexpressionofFasandFasLincreasedinHepG2cells．buttheexpressionofFasLwaslowelthanthatofFas．TheexpressionofFasinsolotransfectionofLIGHTgenegroupiShigherthanthatinthecontrollgroup．andthe combinedtransfectiongroupofLIGHTandIFN．vgenesiShighest(PO．01)．TheexpressionofFasLinthecombinedtransfectionofLIGHT．andIFN．Ywashi2herthanthatinthesolotransfectionofLIGHTonl2hand24h．butwaslowerthanthatinthesolotransfectionofLIGHTon48haftertransfection(尸O．01)．TheapoptosisofHepG2cellsincreasedastime1asted．andtheapoptosisofcombinedtransfectionofLIGHTand IFN．7washigherthanthatofthesolotransfectionofLIGHT(P0．01)．Conclusion：LIGHTandIFN—YgenesSuccessfullytransfectedHepG