【摘要】：胃癌(gastric cancer,GC)和結直腸癌(colorectal cancer,CRC)是我國主要的消化道惡性腫瘤,位居男性腫瘤發(fā)病率的第二和第五位,女性腫瘤發(fā)病率的第三和第四位。結直腸癌發(fā)病率在我國呈上升趨勢,其中超過三分之一的患者會出現(xiàn)肝轉移。目前對于結直腸癌及結直腸癌肝轉移的篩查及監(jiān)測方法包括腸鏡、超聲檢測等等,費用較高,不便于普及。同時,血清標志物檢測的靈敏度和特異度也不理想。尿液作為可以采用非侵入性方式收集的體液,其中含有大量來自于機體的蛋白質,是疾病標志物研究的良好來源。本研究采用定量蛋白質組學策略,將樣本按照疾病進程分為健康對照(healthy individuals,HC)、結直腸癌無轉移、結直腸癌淋巴轉移(CRC with lymph node metastasis,CRC-LNM)以及結直腸癌肝轉移(CRC with liver metastasis,CRC-LM)_4組,每組9例樣本,用10標串聯(lián)質譜標簽(Tandem Mass Tags,TMT)標記,結合二維液相色譜串聯(lián)質譜定量分析結直腸癌、結直腸癌淋巴轉移、結直腸癌肝轉移與健康個體之間蛋白含量的差異。結合質譜數(shù)據(jù)及實驗室前期研究基礎,選擇肝細胞生長因子調節(jié)酪氨酸激酶底物(hepatocyte growth factor-regulated tyrosine kinase substrate,HGS)作為潛在標志物,通過敲降 HGS 觀察結直腸癌細胞系表型的變化。研究發(fā)現(xiàn),利用NC膜富集的方法可以從40mL的正常人和結直腸癌患者尿液中富集到300-800μg的蛋白,對分子量范圍在20-130kDa的蛋白有較好的富集效果。定量蛋白質組學研究發(fā)現(xiàn),四組中共有蛋白995個,差異蛋白富集在細胞凋亡及增殖通路;差異蛋白HGS與細胞遷移能力相關,可能是潛在的疾病標志物。胃癌的癌變過程是多因素長時間共同作用的結果�；蚝铜h(huán)境會影響胃癌患病風險。已有的四項胃癌相關全基因組關聯(lián)研究(genome-wide association study,GWAS)鑒定到 7 個單核苷酸多態(tài)性(single nucleotide polymorphism,SNP)位點,提示遺傳因素在胃癌的發(fā)生發(fā)展過程中有一定的作用。同時,幽門螺桿菌(Helicobacterpylori,H.pylori)感染、吸煙和飲酒也是胃癌相關的重要環(huán)境因素。本研究目的是在胃癌中探索基因-環(huán)境相互作用,研究7個多態(tài)性位點與幽門螺桿菌感染、吸煙和飲酒在胃癌及重度腸上皮化生和不典型增生(severe intestinal metaplasia/dysplasia,severeIM/dysplasia)間的潛在交互作用。研究發(fā)現(xiàn),PSCA rs2294008/rs2976392與幽門螺桿菌感染在胃癌風險上存在顯著的相互作用。同時,PRKAAlrs13361707與幽門螺桿菌感染存在加法相互作用,SLC52A3 rs13042395與飲酒存在加法相互作用。此外,3個SNP位點,MUC1 rs4072037、ZBTB20 rs9841504和PRKAA1 rs13361707與癌前病變相關。提示多態(tài)性位點危險基因型可能與環(huán)境因素,特別是幽門螺桿菌感染和飲酒相互作用,增加胃癌風險。
[Abstract]:Gastric cancer (gastric cancer,GC) and colorectal cancer (colorectal cancer,CRC) are the main malignant tumors of digestive tract in China, ranking the second and fifth in the incidence of cancer in men and the third and fourth in the incidence of tumors in women. The incidence of colorectal cancer is on the rise in China, in which more than 1/3 patients will have liver metastasis. The current screening and monitoring methods for colorectal cancer and colorectal cancer liver metastasis, including endoscopy, ultrasound detection and so on, are expensive and not easy to be popularized. At the same time, the sensitivity and specificity of serum marker detection are not ideal. Urine, as a noninvasive body fluid, contains a lot of proteins from the body and is a good source of disease markers. In this study, quantitative proteomics strategy was used to divide the samples into healthy control (healthy individuals,HC) according to disease progression, no metastasis of colorectal cancer, and (CRC with lymph node metastasis, of lymphatic metastasis of colorectal cancer. CRC-LNM) and (CRC with liver metastasis,CRC-LM _ 4 group, 9 samples from each group were labeled with 10 tandem mass spectrometry tag (Tandem Mass Tags,TMT) and quantitatively analyzed by two dimensional liquid chromatography-tandem mass spectrometry (TLC-MS). Differences in protein content between lymphatic metastasis of colorectal cancer, liver metastasis of colorectal cancer and healthy individuals. Based on the mass spectrometry data and the experimental basis, hepatocyte growth factor regulated tyrosine kinase substrate (hepatocyte growth factor-regulated tyrosine kinase substrate,HGS) was selected as a potential marker to observe the phenotypic changes of colorectal cancer cell lines by knockdown HGS. It was found that the method of NC membrane enrichment could be enriched to 300-800 渭 g protein from the urine of 40mL normal persons and patients with colorectal cancer, and had a good enrichment effect on the proteins with molecular weight range in 20-130kDa. Quantitative proteomics study showed that there were 995 proteins in the four groups, the differential protein was concentrated in apoptosis and proliferation pathway, and differential protein HGS was related to the ability of cell migration, which might be a potential disease marker. The carcinogenesis of gastric cancer is the result of long-term interaction of many factors. Genes and environment can affect the risk of gastric cancer. Seven single nucleotide polymorphisms (single nucleotide polymorphism,SNP) loci have been identified by four gastric cancer associated genome-wide association studies (genome-wide association study,GWAS), suggesting that genetic factors play a role in the development of gastric cancer. At the same time, Helicobacter pylori (Helicobacterpylori,H.pylori) infection, smoking and drinking are also important environmental factors associated with gastric cancer. The aim of this study was to explore the gene-environment interaction in gastric cancer, and to study seven polymorphic sites associated with Helicobacter pylori infection, smoking and alcohol consumption in gastric cancer and severe intestinal metaplasia and atypical hyperplasia of (severe intestinal metaplasia/dysplasia,. Potential interaction between severeIM/dysplasia. A significant interaction between PSCA rs2294008/rs2976392 and Helicobacter pylori infection was found in the risk of gastric cancer. At the same time, there was additive interaction between PRKAAlrs13361707 and Helicobacter pylori infection, and there was additive interaction between SLC52A3 rs13042395 and alcohol consumption. In addition, three SNP loci, MUC1 rs4072037,ZBTB20 rs9841504 and PRKAA1 rs13361707, were associated with precancerous lesions. The results suggested that the risk genotypes of polymorphic loci might interact with environmental factors, especially Helicobacter pylori infection and alcohol consumption, and increase the risk of gastric cancer.
【學位授予單位】：北京協(xié)和醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735
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本文編號：2350819