本文選題：長鏈非編碼RNA + PVT1　； 參考：《青島大學》2017年碩士論文

【摘要】：背景及目的食管癌是一種常見的惡性腫瘤,其發(fā)病率和致死率在中國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慣所造成的。它有兩種病理學類型:食管鱗狀細胞癌和食管腺癌。其中食管鱗癌比較常見。食管癌通常由于患者吞咽困難等不適癥狀,行胃鏡檢查時發(fā)現(xiàn),而此時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,手術不僅會給患者帶來痛苦,也可能產(chǎn)生術后吻合口瘺等并發(fā)癥。因此,早期發(fā)現(xiàn)食管癌并可通過分子機制利用藥物治療食管癌成為一種迫切的需求。長鏈非編碼RNA是一類長約200個核苷酸長度的RNA序列,它雖然沒有編碼蛋白質的作用,但其在胚胎發(fā)育,細胞增殖、分化以及調控基因表達方面發(fā)揮重要的調節(jié)作用。因此推測其在癌癥的發(fā)生和發(fā)展過程中也起到重要的調控作用。PVT1是原癌LncRNA之一,約300bp長,距我們所熟知的MYC基因僅有57kb,由于其特殊的位置關系,推測兩者可能發(fā)揮協(xié)調作用調控腫瘤的發(fā)生發(fā)展。NEAT1是一種長鏈非編碼RNA,位于人類第1號染色體上,與免疫應答,基因表達調控等生物學過程相關,此前已有相關研究證實NEAT1在神經(jīng)膠質瘤、白血病、乳腺癌、喉癌等多種腫瘤中表達上調。目前,對上述兩個LncRNA的研究主要集中與其他腫瘤,與食管癌的關系鮮有報道,因此本文旨在通過檢測長鏈非編碼RNA PVT1和NEAT1在食管鱗狀細胞癌患者腫瘤組織和正常組織的差異,并分析其表達含量和食管癌分期及預后的關系,探討其作為預測食管癌預后和治療的新的分子標志物。方法1長鏈非編碼RNA PVT1和NEAT1在食管鱗狀細胞癌腫瘤組織和配對正常組織的差異性表達研究。1.1收集2015年6月至2016年4月青島大學醫(yī)學院附屬醫(yī)院胸外科54例食管鱗狀細胞癌患者的正常組織(距腫瘤5cm以上)和腫瘤組織,所有患者術前均未接受放化療,術后病理確診為食管鱗狀細胞癌,將切除的新鮮癌組織迅速置于含有RNA保護液的EP管中,并于半小時內放置于-80℃的超低溫冰箱凍存。1.2 Trizol法分別提取食管鱗狀細胞癌腫瘤組織和正常組織中RNA,以微量分光光度計檢測所提取RNA的純度和濃度,取適宜純度(1.80-2.0)之間的RNA用作下一步實驗,不符合要求的重新提取直至符合要求。分別對LncRNA PVT1和LncRNA NEAT1的反轉錄的上、下游設計引物,利用NCBI網(wǎng)站中的gene數(shù)據(jù)庫查找出lnc RNA PVT1和lnc RNA NEAT1的序列全長,使用斯坦福大學的在線引物設計軟件對PVT1基因和NEAT1進行設計,以GAPDH作為對照;后進行反轉錄,通過實時定量聚合酶鏈反應(Real-time PCR)法檢測食管鱗狀細胞癌患者腫瘤組織和正常組織中PVT1和NEAT1的表達含量,結合患者臨床資料,以spss19.0分析其表達含量與患者臨床病理特征之間的關系。2搜集并提取TCGA數(shù)據(jù)庫關于ESCC相關測序資料,利用oncolnc分析此兩種LncRNA表達水平與患者生存之間的關系。結果1長鏈非編碼RNA PVT1和NEAT1在食管鱗狀細胞癌腫瘤組織中的表達高于正常組織,其表達含量的差異具有統(tǒng)計學意義(P0.05),2 LncRNA PVT1和NEAT1的表達水平的高低與患者年齡、性別、腫瘤部位等無明顯相關性(P0.05),而與ESCC的淋巴結是否轉移、腫瘤分化程度、TNM分期等明顯相關(P0.05)3 Kaplan-Meier生存分析顯示:與低表達組相比,PVT1和NEAT1高表達組的5年生存率下降(P0.05)。結論1長鏈非編碼RNA PVT1和NEAT1在食管鱗狀細胞癌腫瘤組織中的表達要明顯高于正常組織。2高表達LncRNA PVT1和NEAT1的ESCC往往分期較晚,預后不良。3 LncRNA PVT1和NEAT1與食管鱗狀細胞癌的發(fā)生發(fā)展密切相關,有可能作為新的分子標志物為食管癌的早期診斷和治療服務。
[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.

【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735.1

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相關期刊論文 前2條

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本文編號：1791619