本文選題：食管癌　切入點(diǎn)：微小RNA-210　出處：《廣西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的微小RNA(microRNA)[1]是一種長(zhǎng)度約為21nt的單鏈非編碼小分子RNA(約22個(gè)核苷酸組成的非編碼小分子RNA)。miRNA在進(jìn)化過程中高度保守,其通過與靶基因序列的3’UTR區(qū)特異性結(jié)合,誘導(dǎo)靶m RNA降解或阻遏其翻譯,miRNA預(yù)計(jì)調(diào)控超過60%的蛋白編碼基因并且參與近乎所有已知的細(xì)胞進(jìn)程。miRNA在細(xì)胞增殖、分化、代謝、凋亡與發(fā)育過程中發(fā)揮重要的調(diào)節(jié)作用。其參與疾病發(fā)生、發(fā)展和轉(zhuǎn)歸過程[2],并通過調(diào)控癌基因或抑癌基因的表達(dá)來影響腫瘤的發(fā)生發(fā)展。近年來,血清腫瘤標(biāo)志物因其獲得相對(duì)比較簡(jiǎn)單、對(duì)病人創(chuàng)傷少,能夠早期提示診斷惡性腫瘤物越來越受到廣大醫(yī)學(xué)研究者青睞。循環(huán)miRNA在血漿和血清中非常穩(wěn)定,它在血清中與特定的蛋白結(jié)合成復(fù)合體后,不會(huì)被RNA裂解酶裂解[3];miRNA在血漿和血清中的穩(wěn)定性使得其實(shí)際值與在病人身上得到的測(cè)試值非常吻合[3]。正因如此,miRNA作為一種腫瘤標(biāo)志物,有極大的潛能。本研究通過收集、提取食管癌患者及正常人血清中miRNA-210,比較觀察組與對(duì)照組之間miRNA-210表達(dá)水平差異,以及不同組織類型、分期、分級(jí)的食管癌患者血清miRNA-210表達(dá)水平差異,探討miRNA-210作為腫瘤標(biāo)志物指導(dǎo)食管癌的早期診斷,以及對(duì)食管癌組織類型、分期、分級(jí)提供診斷依據(jù)。方法經(jīng)前期研究,選取食管癌患者與正常人血清miRNA-210為研究對(duì)象,實(shí)驗(yàn)步驟如下:第一步收集臨床資料,正常對(duì)照組血清樣本取自健康體檢人群(無腫瘤、嚴(yán)重急慢性感染等危及生命基礎(chǔ)病);食管癌患者血清取自廣西醫(yī)科大學(xué)第一附屬醫(yī)院胸外科食管癌患者(食管癌病例的診斷有胃鏡病理活檢或者術(shù)后有病理檢查結(jié)果)。第二步用TRIzol法提取食管癌患者和正常對(duì)照組血清中的miRNA,用實(shí)時(shí)熒光定量PCR法分別檢測(cè)兩組血清miRNA-210水平。第三部,對(duì)比兩組miR-210水平及觀察組miRNA-210與臨床特征的關(guān)系。結(jié)果觀察組miRNA-210,高于對(duì)照組(P0.05),與性別、年齡、病理類型無關(guān)性(P0.05),與TNM分期、組織學(xué)分級(jí)、淋巴結(jié)轉(zhuǎn)移有關(guān)(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),有淋巴轉(zhuǎn)移的高于無淋巴轉(zhuǎn)移的。結(jié)論(1)本研究食管癌患者血清miR-210表達(dá)高于正常人,暗示miR-210升高與食管癌的發(fā)生發(fā)展有關(guān)�？赡芤种埔职┗�,導(dǎo)致食管癌的發(fā)生。(2)食管癌患者血清miR-210與患者TNM分期、組織學(xué)分級(jí)、淋巴結(jié)轉(zhuǎn)移有關(guān),與病理類型無關(guān)。食管癌患者惡性程度越高,其miRNA-210表達(dá)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，

本文編號(hào)：1713193