本文選題：肝癌 + microRNA��； 參考：《吉林大學(xué)》2017年碩士論文

【摘要】：研究背景:肝細(xì)胞癌(Hepatocellular carcinoma,HCC)是成年人最常見(jiàn)的肝臟原發(fā)性惡性腫瘤,占原發(fā)性肝癌的80-90%。在全球范圍內(nèi),其發(fā)病率在人體惡性腫瘤中居第五位,是癌癥相關(guān)死亡的第三大主因。據(jù)統(tǒng)計(jì),HCC發(fā)生率每年增加3-9%,新增病例約78萬(wàn),其中50%發(fā)生在我國(guó),由乙肝導(dǎo)致肝硬化進(jìn)而發(fā)展為HCC是其發(fā)病率逐年遞增的主要原因。HCC的發(fā)生是一個(gè)多因素、多階段的過(guò)程,但由于其發(fā)病隱匿,轉(zhuǎn)移迅速,侵襲性強(qiáng),且缺乏靈敏的早期診斷分子標(biāo)志物,診斷時(shí)多已發(fā)展到晚期。盡管有肝移植、腫瘤切除、栓塞、化療等多種方案適用于HCC的治療,其預(yù)后依然很差,5年生存率不到5%,故尋求高靈敏度、高特異性的腫瘤標(biāo)志物對(duì)提高肝細(xì)胞癌患者的生存質(zhì)量和生存時(shí)間有著極為重要的意義。Micro RNA(miRNA)是一類內(nèi)生的、長(zhǎng)度約為18-25個(gè)核苷酸的非編碼小分子RNA,常通過(guò)結(jié)合靶標(biāo)信使RNA(m RNAs)的3’非翻譯區(qū)(3’UTR)抑制其翻譯或加速其降解這兩種主要機(jī)制調(diào)控m RNA的表達(dá),從而在生物體發(fā)育,疾病的發(fā)生發(fā)展等方面發(fā)揮重要作用,其在腫瘤中的作用也越來(lái)越被人們所重視。研究表明多種miRNA在HCC組織中表達(dá)異常,對(duì)肝癌細(xì)胞的增殖、凋亡、侵襲、轉(zhuǎn)移等發(fā)揮重要的調(diào)控作用,因此分析miRNA在肝癌中的表達(dá)特點(diǎn),闡明miRNA在肝癌中的作用對(duì)HCC診斷和治療具有重要意義。研究材料和方法:本研究通過(guò)對(duì)TCGA數(shù)據(jù)庫(kù)中肝癌組織和癌旁組織miRNA測(cè)序數(shù)據(jù)進(jìn)行分析,篩選出肝癌組織中差異表達(dá)的miRNA并且結(jié)合患者的臨床數(shù)據(jù)確定出與肝癌患者臨床特征相關(guān)的差異表達(dá)miRNA。另外對(duì)與肝癌中明顯異常表達(dá)的miRNA進(jìn)行GO和KEGG Pathway分析。其次,我們挑選出在肝癌組織中明顯高表達(dá)的miR-589-5p對(duì)其在肝癌中的功能進(jìn)行研究,通過(guò)QPCR檢測(cè)其在肝癌細(xì)胞系中的表達(dá)量;在肝癌細(xì)胞中轉(zhuǎn)染miR-589-5p抑制劑后通過(guò)MTT、克隆形成以及流式細(xì)胞周期檢測(cè)研究其對(duì)肝癌細(xì)胞增殖的影響;通過(guò)生物信息學(xué)方法預(yù)測(cè)出miR-589-5p可能的靶基因并通過(guò)QPCR及Western Blot進(jìn)行驗(yàn)證。研究結(jié)果:通過(guò)對(duì)TCGA數(shù)據(jù)庫(kù)中肝癌相關(guān)數(shù)據(jù)的分析,我們總共篩選出49個(gè)在肝癌組織中異常表達(dá)的micro RNA,其中有14個(gè)在肝癌組織中高表達(dá),33個(gè)在肝癌組織中低表達(dá)。結(jié)合患者臨床特征數(shù)據(jù)我們發(fā)現(xiàn)其中有21個(gè)micro RNA與肝癌患者的臨床特征有關(guān)。從這些異常表達(dá)的micro RNA中我們挑選出miR-589-5p研究其在肝癌中的功能,micro RNA測(cè)序數(shù)據(jù)表明,miR-589-5p在肝癌組織中表達(dá)量是在癌旁組織中的1.93倍(P0.001),并且其高表達(dá)與肝癌患者的低生存時(shí)間相關(guān)(P=0.0477);QPCR實(shí)驗(yàn)驗(yàn)證表明miR-589-5p在肝癌細(xì)胞系HepG2和Huh-7中比在肝正常細(xì)胞系L-O2中明顯高表達(dá);抑制HepG2和Huh7中miR-589-5p的表達(dá)后,細(xì)胞的增殖能力降低;通過(guò)生物信息學(xué)方法預(yù)測(cè)出miR-589-5p可能的靶基因MIG-6,用PCR和Western Blot實(shí)驗(yàn)進(jìn)行驗(yàn)證,發(fā)現(xiàn)HepG2細(xì)胞導(dǎo)入miR-589-5p抑制劑后,MIG-6的表達(dá)量明顯升高。研究結(jié)論:1.肝癌組織中存在大量異常表的的micro RNA,其中有些micro RNA的表與患者的臨床特征有關(guān),并且這些micro RNA可能在肝癌中發(fā)揮重要作用。2.MiR-589-5p在肝癌組織和肝癌細(xì)胞中高表達(dá),并且其表達(dá)量與肝癌患者的生存時(shí)間有關(guān),抑制miR-589-5p的表達(dá)后肝癌細(xì)胞的增殖能力下降。3.MiR-589-5p可能是通過(guò)靶向抑制MIG-6的表達(dá)發(fā)揮作用。
[Abstract]:Background: Hepatocellular carcinoma (HCC) is the most common primary liver malignant tumor in adults, which accounts for the 80-90%. of primary liver cancer in the world. Its incidence is the fifth largest in human malignant tumor. It is the third major cause of cancer related death. According to statistics, the incidence of HCC is increased by 3-9%, and the number of new cases is about 78. Ten thousand, 50% of them occur in China. The main cause of the increase of the incidence of liver cirrhosis, which is caused by hepatitis B, is the main cause of its incidence year by year. The occurrence of.HCC is a multi factor, multi stage process. But because of its concealment, rapid metastasis, strong invasiveness, and lack of sensitive early diagnostic markers, the diagnosis of HCC has developed to a late stage. Liver transplantation, tumor resection, embolization, chemotherapy and so on are suitable for the treatment of HCC. The prognosis is still poor and the 5 year survival rate is less than 5%. Therefore, the search for high sensitivity and highly specific tumor markers is of great importance to the improvement of the survival quality and survival time of the patients with hepatocellular carcinoma (.Micro RNA (miRNA) is a kind of endogenous length. RNA, a non coding small molecule of about 18-25 nucleotides, often plays an important role in the development of organisms, the development of the disease and the development of the disease by inhibiting the translation or accelerating its degradation by combining the 3 'non translation zone (3' UTR) of the target messenger RNA (m RNAs) and accelerating its degradation, and thus plays an important role in the development of the organism and the development of the disease, and its role in the tumor is becoming more and more important. The study shows that the expression of a variety of miRNA in HCC tissues plays an important role in regulating the proliferation, apoptosis, invasion and metastasis of hepatoma cells. Therefore, it is important to analyze the expression of miRNA in the liver cancer and to clarify the role of miRNA in the diagnosis and treatment of HCC. After analyzing the miRNA sequencing data of liver cancer tissue and para cancer tissue in the TCGA database, the differential expression of miRNA in the liver cancer tissues was screened and the difference expression related to the clinical features of the patients with liver cancer was determined by combining the clinical data of the patients with miRNA., and the GO and KEGG Pathway analysis of miRNA, which was obviously abnormal in the liver cancer, was analyzed. At the same time, we selected miR-589-5p, which was highly expressed in the liver cancer tissue, to study its function in liver cancer, and to detect its expression in the liver cancer cell line by QPCR. After transfection of miR-589-5p inhibitor in the hepatoma cells, the effects of the cloning and flow cytometry on the proliferation of hepatoma cells were studied. Bioinformatics methods have predicted the possible target genes of miR-589-5p and verified by QPCR and Western Blot. Results: through the analysis of the liver cancer related data in the TCGA database, we screened out 49 abnormal expressions of micro RNA in the liver cancer tissues, of which 14 were highly expressed in the liver cancer tissues and 33 were in the liver cancer tissue. We found that 21 of micro RNA were associated with the clinical characteristics of the patients with liver cancer. We selected miR-589-5p to study the function of miR-589-5p in liver cancer from these abnormal micro RNA. The micro RNA sequencing data showed that the expression of miR-589-5p in the liver cancer tissues was 1.9 in the para cancer tissues. 3 times (P0.001), and its high expression was associated with low survival time of liver cancer patients (P=0.0477); QPCR test showed that miR-589-5p was significantly higher in HepG2 and Huh-7 of liver cancer cell lines than in the normal liver cell line L-O2; the proliferation ability of the cells decreased after inhibiting the expression of miR-589-5p in HepG2 and Huh7; predicted by bioinformatics methods. MIG-6, a possible target gene for miR-589-5p, was verified by PCR and Western Blot experiments. It was found that the expression of MIG-6 was significantly increased after HepG2 cells were introduced into miR-589-5p inhibitors. Conclusions: there are a large number of abnormal tables in micro RNA in 1. liver cancer tissues, and some micro RNA tables are related to the clinical characteristics of the patients. It may play an important role in liver cancer,.2.MiR-589-5p is highly expressed in liver cancer tissues and hepatoma cells, and its expression is related to the survival time of the liver cancer patients. The decrease of the proliferation ability of hepatoma cells after the inhibition of miR-589-5p expression may play a role in the expression of target inhibition of the expression of MIG-6.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.7

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本文編號(hào)：1970045