重组人干扰素λ1基因治疗肺腺癌的实验研究

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1、江苏大学硕士学位论文重组人干扰素λ1基因治疗肺腺癌的实验研究姓名：朱涛峰申请学位级别：硕士专业：内科学指导教师：严玉兰20120601江苏大学硕士学位论文摘要目的：研究重组人IFN．入1对肺腺癌A549细胞皮下移植瘤的影响及其作用机制。方法：建立A549荷瘤裸鼠模型后，随机分为3组，即Ad．IFN．入1治疗组、Ad—LacZ治疗组和PBS对照组。各组每周瘤内注射干预1次，共干预3次。每7天测量肿瘤体积，末次处理1周后小鼠全部处死，绘制肿瘤生长曲线并计算肿瘤干预的抑瘤率，通过采用RT-PCR检测肿瘤内的IFN．入1基因表达，West

2、ernblot检测肿瘤内的IFN．入l的蛋白表达，TUNEL法检测移植瘤的凋亡，流式细胞仪检测脾NK细胞的改变。结果：小鼠感染Ad．IFN．入l后IFN．入1基因在mRNA水平及蛋白水平表达量都明显高于后两组；Ad．IFN．入1组抑瘤率明显高于后两组(p

3、进肿瘤细胞凋亡及刺激体内NK细胞增加。Ad．IFN．入1具有一定的临床应用前景，并有可能成为一种新的治疗肺癌的手段。关键词：IFN．入1；基因治疗；肺腺癌；凋亡；NK细胞江苏大学硕‘士学位论文ABSTRACTObjective：ToinvestigatetheeffectoftherecombinantadenovirushumanIFN．九1onthegrowthofsubcutaneouslytransplantedhumanlungcancercell(A549cellline)innudemiceanditsmechani

4、smofaction．Methods：NudemicemodelscartingA549cellswascreatedanddividedrandomlyintothreegroups(Ad—IFN一九1，Ad—LacZandPBScontrolgroups)．ThenudemiceineachgroupreceivedcorrespondingO．1mLintratumoralinjectioneveryweekandlastedfor3weeks．Thevolumeofthetumorwasmeasuredeveryweeka

5、fterthefirstinjectionandthemicewerekilledatlast．Thegrowthcurvesofthetumorweredrawnandthetumorgrowthinhibitionrateswerecalculated．TheExpressionsoflFN-九1intumortissueofmiceweredetectedbymethodofwestemblotandRT．PCR．Theapoptosisindexofsubcutaneouslytransplantedtumorcellsa

6、ndthechangeofthespleenNKcellswereobservedbytheTUNELassayandtheflowcytometry．Results：TransfectedwiththerecombinantIFN．九1adenovirusvectorinmice，theexpressionoftheIFN-X1geneinthemRNAandproteinlevelsarebothhigherthantheblankandcontrolgroupones．Thetumorgrowthinhibitionrate

7、sofAd—IFN一九lgroupishigherthanthatoftheothertwogroups(p<0．01)，theapoptosisindexandthetransformationofthespleenIII重组人干扰素九1基因治疗肺腺癌的实验研究NKcellsofAd--IFN-·L1grouparealsohigherthantheothertwogroupsQ

8、sfullyinnudemice．TheAd—IFN一九1cansignificantlyinhibitthetumorgrowthinnudemicebyintratumoralinjection，probablybystimulatingthe