本文選題：長(zhǎng)鏈非編碼RNA + PVT1�。� 參考：《青島大學(xué)》2017年碩士論文

【摘要】：背景及目的食管癌是一種常見(jiàn)的惡性腫瘤,其發(fā)病率和致死率在中國(guó)分別列第六位和第八位,食管癌的發(fā)病往往是由不良的飲食習(xí)慣所造成的。它有兩種病理學(xué)類型:食管鱗狀細(xì)胞癌和食管腺癌。其中食管鱗癌比較常見(jiàn)。食管癌通常由于患者吞咽困難等不適癥狀,行胃鏡檢查時(shí)發(fā)現(xiàn),而此時(shí)部分患者已處于晚期。目前食管癌的主要治療手段還是手術(shù),手術(shù)不僅會(huì)給患者帶來(lái)痛苦,也可能產(chǎn)生術(shù)后吻合口瘺等并發(fā)癥。因此,早期發(fā)現(xiàn)食管癌并可通過(guò)分子機(jī)制利用藥物治療食管癌成為一種迫切的需求。長(zhǎng)鏈非編碼RNA是一類長(zhǎng)約200個(gè)核苷酸長(zhǎng)度的RNA序列,它雖然沒(méi)有編碼蛋白質(zhì)的作用,但其在胚胎發(fā)育,細(xì)胞增殖、分化以及調(diào)控基因表達(dá)方面發(fā)揮重要的調(diào)節(jié)作用。因此推測(cè)其在癌癥的發(fā)生和發(fā)展過(guò)程中也起到重要的調(diào)控作用。PVT1是原癌LncRNA之一,約300bp長(zhǎng),距我們所熟知的MYC基因僅有57kb,由于其特殊的位置關(guān)系,推測(cè)兩者可能發(fā)揮協(xié)調(diào)作用調(diào)控腫瘤的發(fā)生發(fā)展。NEAT1是一種長(zhǎng)鏈非編碼RNA,位于人類第1號(hào)染色體上,與免疫應(yīng)答,基因表達(dá)調(diào)控等生物學(xué)過(guò)程相關(guān),此前已有相關(guān)研究證實(shí)NEAT1在神經(jīng)膠質(zhì)瘤、白血病、乳腺癌、喉癌等多種腫瘤中表達(dá)上調(diào)。目前,對(duì)上述兩個(gè)LncRNA的研究主要集中與其他腫瘤,與食管癌的關(guān)系鮮有報(bào)道,因此本文旨在通過(guò)檢測(cè)長(zhǎng)鏈非編碼RNA PVT1和NEAT1在食管鱗狀細(xì)胞癌患者腫瘤組織和正常組織的差異,并分析其表達(dá)含量和食管癌分期及預(yù)后的關(guān)系,探討其作為預(yù)測(cè)食管癌預(yù)后和治療的新的分子標(biāo)志物。方法1長(zhǎng)鏈非編碼RNA PVT1和NEAT1在食管鱗狀細(xì)胞癌腫瘤組織和配對(duì)正常組織的差異性表達(dá)研究。1.1收集2015年6月至2016年4月青島大學(xué)醫(yī)學(xué)院附屬醫(yī)院胸外科54例食管鱗狀細(xì)胞癌患者的正常組織(距腫瘤5cm以上)和腫瘤組織,所有患者術(shù)前均未接受放化療,術(shù)后病理確診為食管鱗狀細(xì)胞癌,將切除的新鮮癌組織迅速置于含有RNA保護(hù)液的EP管中,并于半小時(shí)內(nèi)放置于-80℃的超低溫冰箱凍存。1.2 Trizol法分別提取食管鱗狀細(xì)胞癌腫瘤組織和正常組織中RNA,以微量分光光度計(jì)檢測(cè)所提取RNA的純度和濃度,取適宜純度(1.80-2.0)之間的RNA用作下一步實(shí)驗(yàn),不符合要求的重新提取直至符合要求。分別對(duì)LncRNA PVT1和LncRNA NEAT1的反轉(zhuǎn)錄的上、下游設(shè)計(jì)引物,利用NCBI網(wǎng)站中的gene數(shù)據(jù)庫(kù)查找出lnc RNA PVT1和lnc RNA NEAT1的序列全長(zhǎng),使用斯坦福大學(xué)的在線引物設(shè)計(jì)軟件對(duì)PVT1基因和NEAT1進(jìn)行設(shè)計(jì),以GAPDH作為對(duì)照;后進(jìn)行反轉(zhuǎn)錄,通過(guò)實(shí)時(shí)定量聚合酶鏈反應(yīng)(Real-time PCR)法檢測(cè)食管鱗狀細(xì)胞癌患者腫瘤組織和正常組織中PVT1和NEAT1的表達(dá)含量,結(jié)合患者臨床資料,以spss19.0分析其表達(dá)含量與患者臨床病理特征之間的關(guān)系。2搜集并提取TCGA數(shù)據(jù)庫(kù)關(guān)于ESCC相關(guān)測(cè)序資料,利用oncolnc分析此兩種LncRNA表達(dá)水平與患者生存之間的關(guān)系。結(jié)果1長(zhǎng)鏈非編碼RNA PVT1和NEAT1在食管鱗狀細(xì)胞癌腫瘤組織中的表達(dá)高于正常組織,其表達(dá)含量的差異具有統(tǒng)計(jì)學(xué)意義(P0.05),2 LncRNA PVT1和NEAT1的表達(dá)水平的高低與患者年齡、性別、腫瘤部位等無(wú)明顯相關(guān)性(P0.05),而與ESCC的淋巴結(jié)是否轉(zhuǎn)移、腫瘤分化程度、TNM分期等明顯相關(guān)(P0.05)3 Kaplan-Meier生存分析顯示:與低表達(dá)組相比,PVT1和NEAT1高表達(dá)組的5年生存率下降(P0.05)。結(jié)論1長(zhǎng)鏈非編碼RNA PVT1和NEAT1在食管鱗狀細(xì)胞癌腫瘤組織中的表達(dá)要明顯高于正常組織。2高表達(dá)LncRNA PVT1和NEAT1的ESCC往往分期較晚,預(yù)后不良。3 LncRNA PVT1和NEAT1與食管鱗狀細(xì)胞癌的發(fā)生發(fā)展密切相關(guān),有可能作為新的分子標(biāo)志物為食管癌的早期診斷和治療服務(wù)。
[Abstract]:Background and objective esophageal cancer is a common malignant tumor. Its incidence and mortality rate are sixth and eighth respectively in China. The incidence of esophageal cancer is often caused by bad eating habits. It has two types of pathological types: esophageal squamous cell carcinoma and esophageal adenocarcinoma. In the case of discomfort symptoms such as dysphagia, the patients are found at the time of gastroscopy, while some of the patients are in the late stage. The main treatment for esophageal cancer is still the operation. The operation not only brings pain to the patients, but also may produce postoperative complications such as anastomotic fistula. Therefore, the early detection of esophageal cancer and the use of molecular mechanisms can be used for drug treatment. Cancer of the esophagus is an urgent need. Long chain noncoding RNA is a class of RNA sequences with a length of about 200 nucleotide lengths. Although it does not encode protein, it plays an important regulatory role in embryonic development, cell proliferation, differentiation and regulation of gene expression. Therefore, it is presumed that it is in the process of cancer development and development. It also plays an important regulatory role.PVT1 is one of the primary cancer LncRNA, about 300bp long and only 57kb from the MYC gene we know. Due to its special location, it is presumed that the two may play a coordinated role in regulating the occurrence and development of the tumor..NEAT1 is a long chain non coded RNA, located on the human chromosome first, and the immune response, gene expression. Regulation and other biological processes are related. Previous studies have confirmed that NEAT1 is up regulated in many kinds of tumors, such as glioma, leukemia, breast cancer, and larynx cancer. At present, the two LncRNA studies are mainly focused on other tumors, and the relationship with the cancer of the esophagus is rarely reported. Therefore, the aim of this paper is to detect long chain non coded RNA PVT1 and NE. The difference between the tumor tissue and normal tissue of the patients with esophageal squamous cell carcinoma (AT1), and the relationship between the expression content and the stage of esophageal cancer and the prognosis of the esophagus, are discussed as a new molecular marker for predicting the prognosis and treatment of esophageal cancer. Methods 1 long chain non coding RNA PVT1 and NEAT1 were used in the tumor tissue of the squamous cell carcinoma of the esophagus and the matched normal tissue. Differential expression study.1.1 collected 54 cases of esophageal squamous cell carcinoma in the Department of thoracic surgery of the Affiliated Hospital of Qiingdao University from June 2015 to April 2016. The normal tissues of the patients with esophageal squamous cell carcinoma (more than 5cm) and tumor tissues were collected. All patients were not treated with chemotherapy before operation. The pathological diagnosis of squamous cell carcinoma of the esophagus after operation, and the rapid resection of fresh cancer tissue quickly. In the EP tube containing RNA protection solution, and the cryopreservation.1.2 Trizol method, which was placed at -80 C in half an hour, to extract RNA in the tumor tissues and normal tissues of the squamous cell carcinoma of the esophagus respectively, the purity and concentration of the extracted RNA was detected by a micro spectrophotometer, and RNA between the suitable purity (1.80-2.0) was used as the next experiment. To meet the requirements of RE Extraction until it meets the requirements. The reverse transcription of LncRNA PVT1 and LncRNA NEAT1, downstream design primers, and the gene database in NCBI website are used to find the sequence of LNC RNA PVT1 and LNC RNA NEAT1, and the design software of the online primer design software of the Stanford University is used. As a control, the expression of PVT1 and NEAT1 in tumor tissues and normal tissues of patients with esophageal squamous cell carcinoma was detected by real-time quantitative polymerase chain reaction (Real-time PCR), and the relationship between the expression content and the clinicopathological features of the patients was analyzed by spss19.0 to collect and extract TCGA by spss19.0. The database about ESCC related sequencing data and oncolnc analysis of the relationship between the two LncRNA expressions and the survival of the patients. Results the expression of 1 long chain non coded RNA PVT1 and NEAT1 in the tumor tissues of the squamous cell carcinoma of the esophagus was higher than that of the normal tissue, and the difference of its expression content was statistically significant (P0.05), 2 LncRNA PVT1 and NEAT1. There was no significant correlation between the level of expression and the age, sex, and tumor location of the patients (P0.05), but a significant correlation with the metastasis of ESCC lymph nodes, the degree of tumor differentiation, and the TNM staging (P0.05) 3 Kaplan-Meier survival analysis showed that the 5 year survival rate of the high expression group of PVT1 and NEAT1 decreased (P0.05). Conclusion the 1 long chain non coding group was compared with the low expression group. The expression of RNA PVT1 and NEAT1 in the tumor tissues of squamous cell carcinoma of the esophagus is obviously higher than that of normal tissue.2 high expression LncRNA PVT1 and NEAT1 ESCC often staging later. The poor prognosis.3 LncRNA PVT1 and NEAT1 are closely related to the development and development of squamous cell carcinoma of the esophagus. It may be a new molecular marker for the early diagnosis of esophageal cancer. And treatment services.

【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 Yuwei Zhang;;Epidemiology of esophageal cancer[J];World Journal of Gastroenterology;2013年34期

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本文編號(hào)：1791619