长链非编码RNA CRNDE在人脑胶质瘤中的表达及其临床意义的研究

发布时间：2018-05-21 14:51

本文选题：脑胶质瘤 + CRNDE　；参考：《河北医科大学》2017年硕士论文

【摘要】：目的:脑胶质瘤是多种因素和多类基因相互作用后逐步发展的结果,但它具体的发生和发展机制仍是一个谜团。长链非编码RNA(Lnc RNA)是构成人类基因组中重要的组成部分,虽然其序列上缺乏开放读码框(OFR),不能直接表达蛋白,但它在转录前后以及表观遗传学等多个环节对靶基因进行精准的调控。他通过间接干预基因的表达从而影响到整个生物学的功能。大量研究分析表明,Lnc RNA的异常表达与人类肿瘤的发生、演变有着紧密的关联,而且与肿瘤恶性分化程度以及预后相关。CRNDE是结肠癌差异表达长链非编码RNA,此基因存在于人染色体的反义链上,全长约10kb,具有5个核心外显子和一个附加外显子。它在宫颈癌和直肠癌组织中的表达均明显高于癌旁组织。这种表达差异在脑胶质瘤中是否成立呢?本实验围绕脑胶质瘤组织与瘤旁组织,有关CRDNE表达水平是否存在差异以及CRNDE与肿瘤临床病理学的关联展开研究,通过对两组病人进行长期随访后,获取他们的生存时间并纳入生存分析。其结果可以为脑胶质瘤病的诊断、治疗及预后评估开辟一条新的道路。方法:1收集自2007年至2012年,河北医科大学第二医院手术室,神经外科手术当中取出的胶质瘤标本。经过我们仔细的核查后确认,全部的患者均未曾在术前接受过放射治疗及化学药物治疗。留取进行实验的组织均由病理科鉴定,存储在冻存管内并及时将管放入液氮罐内保存。所收集的80例脑胶质瘤标本参照2007世界卫生组织对中枢神经系统(WHO-CNS)肿瘤的分类标准,其中低级别组42例,高级别组38例。此外,在脑胶质瘤手术当中,收集20例手术入路中显微镜下距瘤组织边缘1cm以内的正常脑组织标本(使用术中导航确定肿瘤边界)作为对照组,收集方法与收集胶质瘤组织一致。2使用实时荧光定量聚合酶链反应(q RT-PCR)方法,获取瘤旁组织与脑胶质瘤组织的CRNDE表达量,并对两者进行比较。采用卡方检验验证CRNDE表达水平与临床病理特征的关系。经过长期随访,获得脑胶质瘤病人的生存时间,采用Kapaln-Meier生存分析,绘制两组患者的生存曲线并对两条曲线进行比较,生存时间从患者手术之日算起,到其死亡或者是末次随访日所经历的时间,单位按月来计数。探索CRNDE表达水平的高低与患者五年生存率的关系。结果:1对比脑胶质瘤组织与瘤旁组织的CRNDE表达水平,脑胶质瘤组织中CRNDE的最后Ct均值为3.89±0.19,瘤旁组织CRNDE的最后Ct的均值为1.00±0.04,因此可以看出,CRNDE在脑胶质瘤的表达水平明显高于瘤旁组织CRNDE的表达水平。对比高级别脑胶质瘤CRNDE的表达水平与低级别组CRNDE的表达水平,可发现高级别肿瘤CRNDE的最后Ct的均值为4.33±0.40,低级别脑胶质瘤CRNDE的最后Ct均值为1.87±0.18,由此可知,高级别脑胶质瘤组织当中CRNDE的表达水平明显高于低级别组。(见图1、图2、表1、表2)2对CRNDE的表达水平与患者年龄、性别、发病形式、肿瘤大小、是否存在坏死、WHO级别、肿瘤的复发,多种临床因素进行比较,使用卡方检验进行分析后表明,CRNDE的表达与肿瘤的复发(P=0.010)、肿瘤的大小(P=0.026)、WHO分级(P=0.003)有关,与年龄(P=0.358)、性别(P=0.204)、发病形式(P=0.171)、是否存在坏死(P=0.367)无关。(见表3)3本组资料中,我们根据CRNDE表达量选取中位表达量来划分高表达组和低表达组。使用Kaplan-Meier进行生存分析显示,高表达组患者的生存时间较低表达组患者明显缩短。比较两者的一年生存率显示:高表达组患者一年生存率为84.4%,低表达组患者一年生存率为97.1%,两者比较有显著差异(P0.05)。比较两组患者五年生存率,发现CRNDE高表达组的脑胶质瘤患者五年生存率仅为15.6%,低表达组患者五年的生存率为37.1%。两者比较有显著差异(P0.05)。(见图3)结论:1在脑胶质瘤组织与瘤旁组织有关CRNDE表达水平的对比中,以及高级别的脑胶质瘤与低级别的脑胶质瘤关于CRNDE表达水平的比较中,我们可以发现CRNDE在脑胶质瘤组织中的表达明显高于瘤旁组织中的表达,且CRNDE的表达水平与WHO分级有关,高级别的脑胶质瘤CRNDE表达水平明显高于低级组。2脑胶质瘤中CRNDE表达水平与肿瘤的大小、WHO分级以及肿瘤是否复发有关,与年龄、性别、发病形式、是否存在坏死因素无关。3脑胶质瘤中CRNDE表达水平越高,往往相同时间内其患者生存率越低,且CRNDE高表达组的患者五年生存率明显小于低表达组。
[Abstract]:Objective: glioma is the result of gradual development of multiple factors and multiple genes interaction, but its specific occurrence and development mechanism is still a mystery. Long chain non coding RNA (Lnc RNA) is an important component of the human genome. Although the sequence lacks the open reading code frame (OFR), it can not express the protein directly, but it is in turn. A number of studies and analyses show that the abnormal expression of Lnc RNA is closely related to the occurrence and evolution of human tumor, and is closely related to the degree of malignant differentiation of the tumor and preconditioning. Post related.CRNDE is a differential expression of long chain noncoding RNA in colon cancer. This gene exists on the antisense chain of human chromosomes, with a total length of about 10KB, with 5 core exons and an additional exon. It is significantly higher in cervical and rectal cancer than in para cancerous tissues. The analysis of the difference in CRDNE expression and the association of CRNDE with the tumor clinicopathology of glioma tissue and paratumoral tissue, and the survival time and survival analysis after long-term follow-up to two groups of patients. The results can be used for the diagnosis, treatment and prognosis evaluation of brain glioma. A new way was established. Methods: 1 the specimens were collected from the operation room of the second hospital of Hebei Medical University from 2007 to 2012. After careful verification, we confirmed that all the patients had not been treated with radiation therapy and chemical therapy before the operation. Scientific identification, stored in the cryopreservation tube and stored in a liquid nitrogen tank in time. 80 specimens of glioma were collected with reference to the classification standard of the 2007 WHO for central nervous system (WHO-CNS) tumor, including 42 in low grade group and 38 in advanced group. In addition, 20 cases of surgical approach were collected during the operation of glioma. The normal brain tissue specimens (using intraoperative navigation to determine the tumor boundary) under the 1cm margin of the tumor tissue (using the intraoperative navigation to determine the tumor boundary) were used as the control group. The collection method was consistent with the collection of glioma tissues and used the real-time fluorescence quantitative polymerase chain reaction (Q RT-PCR) method to obtain the CRNDE expression of the paratumoral tissue and the brain gelatinous tumor tissue, and the two were compared. The relationship between the CRNDE expression level and the clinicopathological features was verified by chi square test. After long-term follow-up, the survival time of the patients with glioma was obtained. The survival curves of the two groups were plotted and compared with the two curves. The survival time was calculated from the day of the hand operation, to the death or the last follow-up day. The relationship between the level of CRNDE expression and the five year survival rate of patients was explored. Results: 1 compared with the expression level of CRNDE in brain glioma tissue and paratumoral tissue, the final Ct mean of CRNDE in glioma tissues was 3.89 + 0.19, and the mean value of final Ct of CRNDE in paratumoral tissue was 1 + 0.04, so it could be seen The expression level of CRNDE in glioma was significantly higher than that of CRNDE in the para tumor tissue. Compared with the expression level of CRNDE and the level of CRNDE in the low level group, the mean value of the final Ct of the high grade tumor CRNDE was 4.33 + 0.40, and the final Ct of the low grade glioma CRNDE was 1.87 + 0.18. The level of CRNDE expression in the advanced glioma tissues was significantly higher than that in the low level group. (see Figure 1, figure 2, table 1, table 2) 2 CRNDE expression levels were compared with patient's age, sex, form, size, necrosis, WHO level, tumor recurrence, multiple clinical factors, and analysis of CRNDE by chi square test. CRNDE The expression is related to tumor recurrence (P=0.010), tumor size (P=0.026), WHO classification (P=0.003), not related to age (P=0.358), sex (P=0.204), pathogenesis (P=0.171) and necrosis (P=0.367). (see Table 3) 3 in the data of this group, we divide the high expression and low expression groups according to the median expression of CRNDE expression. Use Kapla. N-Meier survival analysis showed that the survival time of the high expression group was significantly shortened. The one year survival rate of the two groups showed that the one year survival rate in the high expression group was 84.4%, the one year survival rate in the low expression group was 97.1%, and the difference was significant (P0.05). The five year survival rate of the two groups was compared with that of the two groups. The five year survival rate of the E high expression group was only 15.6%, and the five year survival rate of the low expression group was 37.1%. (P0.05). (see Figure 3) conclusion: 1 in the comparison of the level of CRNDE expression in the brain glioma tissue and the paratumoral tissue, and the high grade glioma and the low grade glioma on CRNDE In the comparison of the expression level, we can find that the expression of CRNDE in the glioma tissue is significantly higher than that in the para tumor tissue, and the expression level of CRNDE is related to the WHO classification. The level of CRNDE expression in the high grade glioma is significantly higher than the level of CRNDE in the.2 glioma of the low-grade group and the size of the tumor, WHO classification and the tumor are the same. The higher the level of CRNDE expression in.3 glioma is not related to age, sex, form of disease, and whether there is a necrosis factor, the lower the survival rate of the patients in the same time, and the five year survival rate in the CRNDE high expression group is significantly smaller than that in the low expression group.
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.41

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