mir-374b-5p对裸鼠人胃癌种植瘤的影响及其靶基因reck的分析word论文.docxVIP

miR-374b-5p对裸鼠人胃癌种植瘤的影响及其靶基因RECK的研究中文摘要研究生：朱蔚远导师：李国庆教授目的:通过裸鼠人胃癌种植瘤模型，探讨miR-374b-5p 及其靶基因RECK 在胃癌生长、侵袭、转移中的作用。方法:1.培养人胃癌细胞株MGC-803，使用BABL/c裸鼠建立人胃癌种植瘤动物模型。2.随机分为3组：空白对照组、阴性对照组(体内转染无义序列，NC-inhibitor)和miR-374b-5p 抑制组(体内转染miR-374b-5p 抑制剂，Hsa-micro-374b-5p inhibitor)，观察肿瘤生长情况，HE 染色观察种植瘤标本的病理学特征。3.采用qRT-PCR检测三组裸鼠体内转染后人胃癌种植瘤组织的miR-374b-5p表达水平；荧光素酶活性检测验证miR-374b-5p 是否通过靶向作用RECK 对其表达进行调控；qRT-PCR、免疫组织化学分别检测三组裸鼠人胃癌种植瘤RECK的mRNA转录水平和蛋白表达情况。结果:1. miR-374b-5p抑制组种植瘤生长较空白对照组和阴性对照组种植瘤生长缓慢。2.miR-374b-5p抑制组的病理学检测提示种植瘤的恶性程度较空白对照组和阴性对照组下降。3.miR-374b-5p抑制组种植瘤的miR-374b-5p表达水平较空白对照组和阴性对照组明显降低。4. 荧光素报告载体系统证实RECK是miR-374b-5p直接调控的靶基因。5. miR-374b-5p抑制组种植瘤的RECK转录水平及蛋白表达水平较空白对照组和2阴性对照组均升高。结论:1. miR-374b-5p 在胃癌中可能扮演着癌基因的角色。2. miR-374b-5p 可能是通过RECK 通路在胃癌的生长、侵袭、转移中发挥了作用。关键词:胃癌；miR-374b-5p；裸鼠；RECK；3EffectofmiR-374b-5pinxenogenichumangastriccancerinnudemiceandtheresearchonitstargetgeneRECKObjective:AbstractBythemodelofxenografthumangastriccancerinnudemicetoexplorethefunction ofmiR-374b-5panditstargetgeneRECKingrowth,invasionandmetastasisofgastric cancer.Methods:CulturedhumangastriccancercelllineMGC-803,usingBABL/cnudemiceto establish an animal model of humangastric cancerxenografts.Randomlydividedintothreegroups:blankcontrolgroup,negativecontrolgroup (internaltransfecionnonsensesequence,NC-inhibitor)andmiR-374b-5psuppression group(internaltransfecionmiR-374b-5pinhibitor,Hsa-micro-374b-5pinhibitor),tumor growth was observed, thepathological featureof tumor was observedbyHE staining.TheexpressionlevelsofmiR-374b-5pinxenogenichumangastriccancerinnude miceafterinternaltransfecionwasdeterminedbyqRT-PCR,whethermiR-374b-5p targetingRECKandregulatingitsexpressionwasdeterminedbyluciferaseactivityassay experiments,themRNAtrascripitionandtheproteinsofRECKwasdeterminedby qRT-PCRand immunohistochemistrystaining.Results:ThegrowthofimplantedtumorinmiR-374b-5psuppressiongroupsignificantly slowlythanthe negativecontrol group andblankcontrol group.Pathologytestindicate thedegreeofmalignancyin miR-374b-5psuppressio