miR-182等在前列腺癌样本中表达的验证及rs2016347位点多态性对结合miR-3175的影响

发布时间：2018-02-14 15:38

本文关键词： 前列腺癌 miRNA IGF1R 多态性位点　出处：《广西医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：第一部分miR-182等在前列腺癌样本中表达的验证目的:本研究通过研究micro RNA联合筛查与前列腺癌诊断的关系,为探索前列腺癌发生发展的可能生物学机制提供理论依据,同时为筛选前列腺癌相关可能的潜在生物分子标记物及优化临床诊断的实施提供更有效的指导方法:用RT-qPCR对前期筛选出来的五个miRNA在前列腺癌组织中和癌旁组织中的表达进行测定,然后用SPSS软件对这五个miRNA进行联合诊断分析。结果:通过RT-qPCR对前期数据挖掘分析出来的有五个miRNA的表达进行测定,我们发现miR-182-3p、miR-182-5p、miR-200c在前列腺癌症组织中相对于癌旁组织表达上调,miR-182-5p上调最明显(p=0.0018),其次是miR-200c(p=0.0031)和miR-182-3p(p=0.005);miR-211-3p和miR-221-5p在前列腺癌症组织中相对于癌旁组织表达下调,miR-221-3p下调最明显(p=0.0088),其次是miR-221-5p(p=0.0319)。五个miRNA进行ROC联合诊断得到的AUC曲线下面积为1。结论:我们研究发现在中国人群前列腺癌样本中,miR-182-3p,miR-182-5p,miR-200c的表达相对于癌旁组织上调,并且miR-182-5p上调最显著;miR-221-3p和miR-221-5p的表达下调,下调最显著的是miR-221-3p。通过ROC联合诊断进一步说明这五个miRNA有可能作为前列腺癌联合筛查的标志物。第二部分rs2016347位点多态性对结合miR-3175的影响目的:为了探究位于IGF1R基因3'UTR与前列腺癌(prostate cancer,PCa)相关的多态性位点rs2016347通过改变与miRNA结合影响PCa的风险。方法:本研究用miRbase等生物信息学工具预测靶向miRNA,然后用热力学模型方法计算结合能情况,最后用双荧光素酶报告基因技术对HEK293T在体外检测改变rs2016347位点对靶向miRNA结合的影响。结果:miRbase等工具预测结果显示,miR-3175与IGF1R的结合区位于多态性位点rs2016347。热力学模型方法计算结果表明,相对位点G,hsa-miR3175与IGF1R的3'UTR端在rs2016347位点T上更能有效结合。双荧光素酶报告基因的检测结果显示,miR-3175能与带有rs2016347等位位点(G或T)IGF1R 3'UTR结合,miR-3175与等位位点T的结合更稳定,miR-3175与IGF1R基因的结合起到稳定IGF1R基因表达的作用。结论:多态性位点rs2016347等位位点T在PCa的发展中风险性更大。
[Abstract]:The purpose of this study is to study the relationship between micro RNA combined screening and prostate cancer diagnosis in order to provide theoretical basis for exploring the possible biological mechanism of prostate cancer. It also provides more effective guidance for screening potential biomolecules associated with prostate cancer and for optimizing clinical diagnosis by using RT-qPCR to detect five pre-screened miRNA in prostate cancer tissues and adjacent tissues. To determine the expression of. Then the five miRNA were diagnosed and analyzed by SPSS software. Results: the expression of five miRNA was detected by RT-qPCR. We found that the upregulation of miR-182-182-5pmmiR-200c in prostate cancer tissues was the most obvious relative to the expression of miR-182-5p, followed by miR-200ctrop 0.0031) and the down-regulation of miR-221-3p and miR-221-5p expression in prostate cancer tissues, followed by the down-regulation of miR-221-3p in prostate cancer tissues, followed by the down-regulation of miR-221-3p expression in prostate cancer tissues, followed by the down-regulation of miR-221-3p expression in prostate cancer tissues, followed by the down-regulation of miR-182-3p and miR-221-3p, followed by the down-regulation of miR-211-3p and miR-221-3p expression in prostate cancer tissues. The area under the AUC curve obtained from the combined diagnosis of five miRNA with ROC was 1.Conclusion: we found that the expression of miR-182-3pmmiR-182-5pmmiR-200c was up-regulated in Chinese prostate cancer samples as compared with adjacent tissues. MiR-182-5p up-regulated the expression of miR-221-3p and miR-221-5p, and down-regulated the expression of miR-221-3p and miR-221-5p. The most significant down-regulation was miR-221-3p.Through the combined diagnosis of ROC, it was further demonstrated that these five miRNA might be used as markers for combined screening of prostate cancer. Part two: the effect of rs2016347 locus polymorphism on binding miR-3175: to explore the location of IGF1R gene. Rs2016347, a polymorphic site associated with prostate cancer, affects the risk of PCa by changing its binding to miRNA. Methods: in this study, the targeted miRNAs were predicted by bioinformatics tools such as miRbase, and the binding energy was calculated by thermodynamics model. Finally, double luciferase reporter gene technique was used to detect the effect of changing rs2016347 sites on the binding of HEK293T to targeted miRNA in vitro. Results the results of prediction of the binding region of IGF1R to IGF1R showed that the binding region was located at the polymorphic site rs20163477.The thermodynamic model method showed that the binding region of RMR-3175 was located at the polymorphic site rs2016347. The relative locus Gnhsa-miR3175 can bind more effectively to the 3UTR terminal of IGF1R on the rs2016347 site T. the detection results of the double luciferase reporter gene show that the mmiR-3175 can bind more stably to the allelic T with the rs2016347 allele G or TIGF1R3175. Conclusion: the polymorphic rs2016347 allele T plays a more important role in the development of PCa.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.25

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