血清MicroRNA作为预测多形性胶质母细胞瘤患者预后的生物标志物

发布时间：2018-03-13 10:48

本文选题：微小RNA　切入点：多形性胶质母细胞瘤　出处：《天津医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：多形性胶质母细胞瘤(GBM)是在成人中最常见的原发性中枢神经系统肿瘤,发病率为3/10万。尽管近几十年来肿瘤学已经取得了极大的进展,通过手术、放疗、化疗等一系列综合治疗手段治疗后多形性胶质母细胞瘤患者的预后仍然不容乐观,中位生存时间为14个月,复发的可能性几乎为100%,且在相同方案治疗下,并不是所有的患者都能获得相同的治疗效果。已经证实的与预后相关的因素非常有限,包括:年龄、KPS、mini-mental status examination score、肿瘤切除率和MGMT启动子是否甲基化。目前,虽然分子和基因表达谱的研究已经鉴别出能够影响GBM患者预后的因素,但是这些因素依然存在着这样那样的缺点。Micro RNA(mi RNA)是一种内源性的非蛋白编码的小分子RNA,它可以在转录后水平上对靶基因的表达进行负性调控。mi RNA的异常表达与很多种疾病都息息相关,尤其是肿瘤。由于单个的mi RNA就能调节数以百计的下游基因,故与蛋白表达基因普相比,应用mi RNA表达谱会获得更多且更精准的关于癌症分类方面的信息。目前已经有一些研究筛选出GBM肿瘤组织中的mi RNA表达谱作为预测GBM的预后因子。近年来,随着血清mi RNAs的发现,mi RNAs的研究领域聚焦在血清mi RNAs与疾病的诊断与预后上。作为新的生物学标志物,血清mi RNAs有着快捷,方便,准确性较高等优点。已经有研究证实,血清mi RNAs可以作为预测非小细胞肺癌预后的生物标志物。然而,目前没有任何研究是应用血清mi RNA表达谱筛选的方法来预测GBM患者的生存期的。在本研究中,我们假设GBM患者血清中存在一种特定的mi RNA表达谱可以作为预测预后的指纹。为了证明这一假设,我们应用Solexa(Solexa Sequencing)技术、q RT-PCR(Quantitative Real Time Polymerase Chain Reaction)等实验技术和IBM SPSS 19.0软件进行统计学分析。最后得出结论。首先,我们从北京天坛医院收集了127例GBM患者的血清标本,同时对这些患者进行长期的随访,将患者血清标本按照生存期的长短分为两组(以中位生存期393天为界)。接下来,在两组标本中各选出20例,每例血清取1ml混为两种血清样本,并提取总RNA,通过Solexa Sequencing技术检测两种血清标本中差异表达的mi RNAs。结果显示,有63个mi RNAs表达量在两种血清标本中存在差异。随后,在差异表达的mi RNAs中,根据表达差异的大小、已有的文献报道和本实验室的研究成果,选出了16个mi RNAs作为下一步验证的目标。然后根据q RT-PCR(quantitive real time polymerase chain reaction)技术对选出的这些mi RNAs的表达水平做逐一验证,根据表达差异在2倍以上且T-test结果P值0.05的具有稳定表达差异的mi RNAs,即:mi R-27b和mi R-148b。最后我们分别对mi R-27b和mi R-148b评估GBM预后的敏感性与特异性做了ROC曲线,曲线下面积分别为:0.872和0.818我们应用SPSS19.0中的Kaplan-Meier方法和Cox regression对这些mi RNAs做了统计学分析。结果显示GBM患者血清中mi RNAs的表达水平是独立于年龄与性别的预后的相关因素,且与预后成负相关关系。
[Abstract]:Glioblastoma multiforme (GBM) is the most common in adults in primary central nervous system tumors, the incidence rate of 3/10 million. Although in recent decades oncology has made great progress, by surgery, radiotherapy, chemotherapy and a series of comprehensive treatment after the prognosis of patients with glioma of shape the tumor cells are still not optimistic, the median survival time was 14 months, the possibility of recurrence is almost 100%, and in the same treatment, not all patients can get the same treatment. Proven and prognosis related factors is very limited, including: age, KPS, Mini-Mental Status Examination score the resection rate of tumor, and MGMT promoter methylation. At present, although the molecular and gene expression studies have identified factors can affect the prognosis of patients with GBM, but these factors still exist such as .Micro RNA (MI RNA) disadvantage is the small molecule RNA non protein encoding an endogenous, it can be carried out on the expression of target genes at the post transcriptional level of abnormal expression of negative regulation of.Mi RNA and many kinds of diseases are closely related, especially tumor. Because a single Mi downstream gene RNA can regulate hundreds of therefore, gene and protein expression of Pu compared to application of MI RNA expression profiles will get more and more information about the cancer classification precision. At present there have been some studies selected from the MI RNA GBM in tumor tissue expression as a prognostic factor of GBM. In recent years, along with the level of MI RNAs, MI RNAs the research focused on the diagnosis and prognosis of serum mi RNAs and disease. As a new biological marker, serum mi RNAs is a fast, convenient, the advantages of high accuracy. Studies have shown that serum mi RNAs can be To predict the prognosis of non - small cell lung cancer biomarker. However, there is no study on expression of serum mi RNA spectrum screening method to predict the survival of patients with GBM. In this study, we assume that there is a mi RNA specific expression profiles can be used to predict the prognosis of the fingerprint in the serum of patients with GBM. In order to prove this hypothesis, we used Solexa (Solexa Sequencing), Q RT-PCR (Quantitative Real Time Polymerase Chain Reaction) and other experimental technology and IBM SPSS 19 software for statistical analysis. Finally draw the conclusion. First, we collected from Beijing Tiantan Hospital, 127 serum samples from GBM patients, and the long-term follow-up of these patients the serum samples of patients, according to the length of survival were divided into two groups (with a median survival of 393 days for the sector). Then, in the two groups were selected from each of 20 cases, 1 cases of each serum Ml mixed into two serum samples, and total RNA was extracted and detected by Solexa Sequencing technique two differentially expressed in serum samples of MI RNAs. showed that the expression of 63 mi RNAs in two serum samples of variance. Then, differences in the expression of MI in RNAs, according to the differential expression of the size of existing the literature reports and the laboratory research results, selected 16 mi RNAs as the next test target. Then according to the Q RT-PCR (quantitive real time polymerase chain reaction) expression levels for the selected Mi these RNAs do one by one verification, according to the difference of expression in more than 2 times and the results of T-test P 0.05 with a stable expression mi RNAs, namely: Mi R-27b and MI R-148b. respectively at the end of the MI R-27b and MI R-148b to evaluate the prognosis of GBM the sensitivity and specificity of the ROC curve, the area under the curve were 0.872 and 0.818 I We used SPSS19.0's Kaplan-Meier method and Cox regression to do statistical analysis of these mi RNAs. The results showed that MI RNAs expression level in GBM patients was independent of age and sex prognosis, and negatively correlated with prognosis.

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.41

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