miR-210在食管癌患者血清中的表达及临床意义

发布时间：2018-04-05 04:23

  本文选题：食管癌　切入点：微小RNA-210　出处：《广西医科大学》2017年硕士论文

【摘要】：目的微小RNA(microRNA)[1]是一种长度约为21nt的单链非编码小分子RNA(约22个核苷酸组成的非编码小分子RNA)。miRNA在进化过程中高度保守,其通过与靶基因序列的3’UTR区特异性结合,诱导靶m RNA降解或阻遏其翻译,miRNA预计调控超过60%的蛋白编码基因并且参与近乎所有已知的细胞进程。miRNA在细胞增殖、分化、代谢、凋亡与发育过程中发挥重要的调节作用。其参与疾病发生、发展和转归过程[2],并通过调控癌基因或抑癌基因的表达来影响肿瘤的发生发展。近年来,血清肿瘤标志物因其获得相对比较简单、对病人创伤少,能够早期提示诊断恶性肿瘤物越来越受到广大医学研究者青睐。循环miRNA在血浆和血清中非常稳定,它在血清中与特定的蛋白结合成复合体后,不会被RNA裂解酶裂解[3];miRNA在血浆和血清中的稳定性使得其实际值与在病人身上得到的测试值非常吻合[3]。正因如此,miRNA作为一种肿瘤标志物,有极大的潜能。本研究通过收集、提取食管癌患者及正常人血清中miRNA-210,比较观察组与对照组之间miRNA-210表达水平差异,以及不同组织类型、分期、分级的食管癌患者血清miRNA-210表达水平差异,探讨miRNA-210作为肿瘤标志物指导食管癌的早期诊断,以及对食管癌组织类型、分期、分级提供诊断依据。方法经前期研究,选取食管癌患者与正常人血清miRNA-210为研究对象,实验步骤如下:第一步收集临床资料,正常对照组血清样本取自健康体检人群(无肿瘤、严重急慢性感染等危及生命基础病);食管癌患者血清取自广西医科大学第一附属医院胸外科食管癌患者(食管癌病例的诊断有胃镜病理活检或者术后有病理检查结果)。第二步用TRIzol法提取食管癌患者和正常对照组血清中的miRNA,用实时荧光定量PCR法分别检测两组血清miRNA-210水平。第三部,对比两组miR-210水平及观察组miRNA-210与临床特征的关系。结果观察组miRNA-210,高于对照组(P0.05),与性别、年龄、病理类型无关性(P0.05),与TNM分期、组织学分级、淋巴结转移有关(P0.05),Ⅳ期高于Ⅲ期,Ⅲ期高于(Ⅰ-Ⅱ)期(P0.05),低分化高于中分化,中分化高于高分化(P0.05),有淋巴转移的高于无淋巴转移的。结论(1)本研究食管癌患者血清miR-210表达高于正常人,暗示miR-210升高与食管癌的发生发展有关。可能抑制抑癌基因,导致食管癌的发生。(2)食管癌患者血清miR-210与患者TNM分期、组织学分级、淋巴结转移有关,与病理类型无关。食管癌患者恶性程度越高,其miRNA-210表达水平越高。
[Abstract]:Objective small RNAs [1] are single-stranded, non-coding, small RNAs with a length of about 21nt (about 22 nucleotides) that are highly conserved during evolution and specifically bound to the 3'UTR region of the target gene sequence.Target RNA is induced to degrade or inhibit the translation of more than 60% of the protein encoding genes and participate in almost all known cellular processes. MiRNAs play an important role in cell proliferation, differentiation, metabolism, apoptosis and development.It is involved in the process of occurrence, development and prognosis of the disease [2], and affects the occurrence and development of tumor by regulating the expression of oncogene or tumor suppressor gene.In recent years, serum tumor markers are more and more popular among medical researchers because of their relatively simple acquisition and less trauma to patients.Circulating miRNA is very stable in plasma and serum, and it binds to a specific protein in the serum to form a complex.The stability of [3] miRNAs not lysed by RNA lyase in plasma and serum makes the actual values very consistent with the values obtained in patients [3].This is why miRNA has great potential as a tumor marker.In this study, we extracted miRNA-210 from the serum of esophageal cancer patients and normal people. The difference of miRNA-210 expression between the observation group and the control group, and the difference of serum miRNA-210 expression in different tissue types, stages and grades of esophageal cancer patients were compared.To explore the early diagnosis of esophageal carcinoma with miRNA-210 as a tumor marker, and to provide diagnostic basis for histological type, staging and grading of esophageal carcinoma.Methods Serum miRNA-210 of esophageal cancer patients and normal subjects was selected as the study object after previous study. The experimental steps were as follows: the first step was to collect clinical data. The serum samples of normal control group were taken from healthy people (no tumor).The serum of patients with esophageal cancer was obtained from thoracic surgery of the first affiliated hospital of Guangxi Medical University.In the second step, TRIzol method was used to extract miRNAs from the serum of esophageal cancer patients and normal controls, and real-time fluorescence quantitative PCR was used to detect the serum miRNA-210 levels in the two groups.In the third part, the level of miR-210 in the two groups and the relationship between miRNA-210 and clinical features in the observation group were compared.Results the miRNA-210 in the observation group was higher than that in the control group (P 0.05). It was not related to sex, age and pathological type. It was related to TNM stage, histological grade, lymph node metastasis, stage 鈪，

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