本文選題：核酸切除修復(fù)基因 + 遺傳變異�。� 參考：《南京中醫(yī)藥大學(xué)》2017年碩士論文

【摘要】：背景:結(jié)直腸癌(包括結(jié)腸癌與直腸癌)是世界上常見的消化系統(tǒng)惡性腫瘤之一,在我國,結(jié)直腸癌的發(fā)病率目前已呈逐年上升趨勢,嚴重威脅著人類的健康。盡管目前篩查早期結(jié)直腸癌的方法為數(shù)不少,但由于結(jié)直腸癌起病隱匿,大部分患者確診時已發(fā)展至中晚期,無法進行手術(shù)治療,因而輔助化療已成為結(jié)直腸癌治療中不可或缺的手段之一。奧沙利鉑(oxaliplatin)是第三代鉑類化合物,是目前針對中晚期結(jié)直腸癌化療聯(lián)合方案的標準藥物之一,其主要原理是通過與脫氧核糖核酸雙鏈共價結(jié)合形成鏈間交聯(lián),阻斷腫瘤細胞DNA的復(fù)制和轉(zhuǎn)錄,從而發(fā)揮抗腫瘤作用。核酸切除修復(fù)系統(tǒng)(nucleotide excision repair,NER)是DNA修復(fù)的基本通路,修復(fù)紫外線所致嘧啶二聚體、脫氧核糖核酸加合物、光化合物等累積性損傷,是清除大劑量鉑類藥所引起的脫氧核糖核酸損傷的重要系統(tǒng)。人類基因組計劃已經(jīng)確認,基因的遺傳與變異會影響基因的結(jié)構(gòu)、功能以及表達,進而改變機體的生物學(xué)功能。單核苷酸多態(tài)性(single nucleotide polymorphisms,SNPs)是最常見的基因遺傳變異。能夠改變基因結(jié)構(gòu)或調(diào)節(jié)基因表達量,從而調(diào)控機體的作用。研究發(fā)現(xiàn),單個SNP作用或多個SNPs間的聯(lián)合作用與腫瘤發(fā)生發(fā)展及預(yù)后顯著相關(guān)。耐藥現(xiàn)象主要產(chǎn)生于修復(fù)系統(tǒng)清除藥物,這是減弱化療藥物療效的主要原因,而DNA修復(fù)基因的SNP密切影響著與修復(fù)系統(tǒng)的修復(fù)能力。迄今為止,已有多篇DNA修復(fù)通路SNPs與腫瘤及結(jié)直腸癌發(fā)病風(fēng)險和化療敏感性研究的報道。大量研究結(jié)果證實,NER通路與結(jié)直腸癌化療敏感性以及生存預(yù)后存在潛在的關(guān)聯(lián)。方法:本課題通過對166例結(jié)直腸癌患者的臨床病例研究,篩選NER通路中可能存在的重要基因遺傳變異,統(tǒng)計其與結(jié)直腸癌化療敏感性及生存預(yù)后的關(guān)聯(lián),尋找到相關(guān)的遺傳變異rs5030740及所在基因RPA1。在此基礎(chǔ)上,通過分子生物學(xué)方法研究關(guān)鍵基因RPA1及遺傳變異rs5030740對腫瘤細胞的表型影響,進而揭示其在奧沙利鉑化療敏感性及生存預(yù)后中的可能機制,為預(yù)測結(jié)直腸癌化療敏感性及進行個體化治療提供理論依據(jù)。結(jié)果:對166個以奧沙利鉑為主要治療方式的結(jié)直腸癌患者進行化療敏感性及生存預(yù)后與RPA1遺傳變異的關(guān)聯(lián)性分析,尋找到相關(guān)的遺傳變異rs5030740及所在基因RPA1。課題發(fā)現(xiàn)rs5030740位點T等位基因突變到C等位基因與結(jié)直腸癌患者奧沙利鉑化療敏感性降低有關(guān),并且結(jié)直腸癌患者用藥后無進展生存期縮短有關(guān)。此外,本課題組通過網(wǎng)站預(yù)測,數(shù)據(jù)庫分析,細胞學(xué)實驗等驗證了 rs5030740位點的突變導(dǎo)致了 miRNA-let-7e-5p與PPA1基因3'-UTR的結(jié)合能力的降低,促使RPA1的表達量上升。最后,課題組通過一系列細胞表型實驗(細胞增殖、周期及凋亡)進一步揭示了RPA1的表達量升高可增加腫瘤細胞的增殖能力,減少細胞凋亡數(shù)目,而這一現(xiàn)象在奧沙利鉑藥物的刺激下,更為明顯。結(jié)論:基因rs5030740的突變可能導(dǎo)致miRNA-let-7e-5p的結(jié)合能力減弱,RPA1的異常高表達,進而通過增加腫瘤細胞的增殖力及減少凋亡率來延長腫瘤的生存能力,最終影響了奧沙利鉑化療藥物的效果及結(jié)直腸癌患者的生存預(yù)后。該研究提示了 RPA1基因rs5030740多態(tài)性可作為中國結(jié)直腸癌人群奧沙利鉑療效及預(yù)后的重要的潛在生物學(xué)指標。
[Abstract]:Background: colorectal cancer (including colon and rectal cancer) is one of the most common digestive malignant tumors in the world. In China, the incidence of colorectal cancer is now increasing year by year, which seriously threatens human health. Although there are many methods of screening colorectal cancer at the early stage, the majority of colorectal cancer are hidden and most of them are hidden. The patient has been developed to the middle and late stages and can not be operated on. Therefore, adjuvant chemotherapy has become one of the indispensable means in the treatment of colorectal cancer. Oxaliplatin (oxaliplatin) is the third generation of platinum compounds, which is one of the standard drugs for the combination of advanced colorectal cancer chemotherapy. The double chain covalent binding of oxyribonucleic acid forms interchain crosslinking to block the replication and transcription of tumor cell DNA and thus plays an antitumor effect. The nucleic acid excision repair system (nucleotide excision repair, NER) is the basic pathway of DNA repair, and repairs the cumulative damage of UV induced pyrimidine two polymer, deoxyribonucleic acid adducts, and light compounds. It is an important system for deoxyribonucleic acid damage caused by large doses of platinum drugs. The human genome project has confirmed that genetic and mutation of genes affect the structure, function, and expression of genes, and then change the biological function of the body. Single nucleotide polymorphisms (SNPs) is the most common gene. Genetic variation. It can change the structure of the gene or regulate the amount of gene expression, which regulates the role of the body. Studies have found that a single SNP action or a combination of multiple SNPs interactions is significantly related to the development and prognosis of the tumor. The SNP of the complex gene is closely related to the repair ability of the repair system. So far, there have been several reports of the DNA repair pathway SNPs and the risk of cancer and colorectal cancer and chemosensitivity. A large number of results have shown that there is a potential association between the NER pathway and the chemotherapy sensitivity and survival of colorectal cancer. After studying the clinical case study of 166 patients with colorectal cancer, we screened the important genetic variation that may exist in the NER pathway, statistics its association with the chemosensitivity of colorectal cancer and the survival prognosis. On the basis of finding the related genetic variation rs5030740 and the gene RPA1., the key gene RPA1 is studied by molecular biology method. And the effect of genetic variation rs5030740 on the phenotype of tumor cells, and then reveal its possible mechanism in the chemotherapeutic sensitivity and survival of oxaliplatin, providing a theoretical basis for predicting chemotherapy sensitivity and individualized treatment of colorectal cancer. Results: chemotherapy for 166 colorectal cancer patients with oxaliplatin as the main treatment method The correlation analysis of sensitivity, survival prognosis and RPA1 genetic variation, finding the related genetic variation rs5030740 and the gene RPA1., found that the mutation of the rs5030740 loci T allele to the C allele was associated with the reduction of oxaliplatin chemosensitivity in colorectal cancer patients, and the non progression survival period of colorectal cancer patients was reduced. In addition, the research group confirmed that the mutation of rs5030740 loci led to the decrease of the binding capacity of miRNA-let-7e-5p and PPA1 gene 3'-UTR by site prediction, database analysis, and cytological experiments. Finally, the group through a series of cell phenotypic experiments (cell proliferation, cycle and apoptosis) into one. The increase in the expression of RPA1 can increase the proliferation of tumor cells and reduce the number of apoptotic cells. This phenomenon is more obvious under the stimulation of oxaliplatin. Conclusion: the mutation of the gene rs5030740 may lead to the weakening of the binding capacity of miRNA-let-7e-5p, the abnormal high expression of RPA1, and the increase of the tumor cells. The effects of oxaliplatin on the survival of the tumor and the survival of the patients with colorectal cancer are ultimately affected by colonization and reducing the rate of apoptosis. This study suggests that the RPA1 gene rs5030740 polymorphism can be used as a major potential biological indicator of the efficacy and prognosis of oxaliplatin in Chinese colorectal cancer population.
【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R735.34

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本文編號：2057833