本文關(guān)鍵詞： 前列腺癌 miRNA IGF1R 多態(tài)性位點(diǎn)　出處：《廣西醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：第一部分miR-182等在前列腺癌樣本中表達(dá)的驗(yàn)證目的:本研究通過(guò)研究micro RNA聯(lián)合篩查與前列腺癌診斷的關(guān)系,為探索前列腺癌發(fā)生發(fā)展的可能生物學(xué)機(jī)制提供理論依據(jù),同時(shí)為篩選前列腺癌相關(guān)可能的潛在生物分子標(biāo)記物及優(yōu)化臨床診斷的實(shí)施提供更有效的指導(dǎo)方法:用RT-qPCR對(duì)前期篩選出來(lái)的五個(gè)miRNA在前列腺癌組織中和癌旁組織中的表達(dá)進(jìn)行測(cè)定,然后用SPSS軟件對(duì)這五個(gè)miRNA進(jìn)行聯(lián)合診斷分析。結(jié)果:通過(guò)RT-qPCR對(duì)前期數(shù)據(jù)挖掘分析出來(lái)的有五個(gè)miRNA的表達(dá)進(jìn)行測(cè)定,我們發(fā)現(xiàn)miR-182-3p、miR-182-5p、miR-200c在前列腺癌癥組織中相對(duì)于癌旁組織表達(dá)上調(diào),miR-182-5p上調(diào)最明顯(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在前列腺癌癥組織中相對(duì)于癌旁組織表達(dá)下調(diào),miR-221-3p下調(diào)最明顯(p=0.0088),其次是miR-221-5p(p=0.0319)。五個(gè)miRNA進(jìn)行ROC聯(lián)合診斷得到的AUC曲線下面積為1。結(jié)論:我們研究發(fā)現(xiàn)在中國(guó)人群前列腺癌樣本中,miR-182-3p,miR-182-5p,miR-200c的表達(dá)相對(duì)于癌旁組織上調(diào),并且miR-182-5p上調(diào)最顯著;miR-221-3p和miR-221-5p的表達(dá)下調(diào),下調(diào)最顯著的是miR-221-3p。通過(guò)ROC聯(lián)合診斷進(jìn)一步說(shuō)明這五個(gè)miRNA有可能作為前列腺癌聯(lián)合篩查的標(biāo)志物。第二部分rs2016347位點(diǎn)多態(tài)性對(duì)結(jié)合miR-3175的影響目的:為了探究位于IGF1R基因3'UTR與前列腺癌(prostate cancer,PCa)相關(guān)的多態(tài)性位點(diǎn)rs2016347通過(guò)改變與miRNA結(jié)合影響PCa的風(fēng)險(xiǎn)。方法:本研究用miRbase等生物信息學(xué)工具預(yù)測(cè)靶向miRNA,然后用熱力學(xué)模型方法計(jì)算結(jié)合能情況,最后用雙熒光素酶報(bào)告基因技術(shù)對(duì)HEK293T在體外檢測(cè)改變r(jià)s2016347位點(diǎn)對(duì)靶向miRNA結(jié)合的影響。結(jié)果:miRbase等工具預(yù)測(cè)結(jié)果顯示,miR-3175與IGF1R的結(jié)合區(qū)位于多態(tài)性位點(diǎn)rs2016347。熱力學(xué)模型方法計(jì)算結(jié)果表明,相對(duì)位點(diǎn)G,hsa-miR3175與IGF1R的3'UTR端在rs2016347位點(diǎn)T上更能有效結(jié)合。雙熒光素酶報(bào)告基因的檢測(cè)結(jié)果顯示,miR-3175能與帶有rs2016347等位位點(diǎn)(G或T)IGF1R 3'UTR結(jié)合,miR-3175與等位位點(diǎn)T的結(jié)合更穩(wěn)定,miR-3175與IGF1R基因的結(jié)合起到穩(wěn)定IGF1R基因表達(dá)的作用。結(jié)論:多態(tài)性位點(diǎn)rs2016347等位位點(diǎn)T在PCa的發(fā)展中風(fēng)險(xiǎn)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.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R737.25

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本文編號(hào)：1511033