消化道肿瘤的尿蛋白标志及遗传—环境交互作用研究
发布时间：2018-11-23 07:45
【摘要】：胃癌(gastric cancer,GC)和结直肠癌(colorectal cancer,CRC)是我国主要的消化道恶性肿瘤,位居男性肿瘤发病率的第二和第五位,女性肿瘤发病率的第三和第四位。结直肠癌发病率在我国呈上升趋势,其中超过三分之一的患者会出现肝转移。目前对于结直肠癌及结直肠癌肝转移的筛查及监测方法包括肠镜、超声检测等等,费用较高,不便于普及。同时,血清标志物检测的灵敏度和特异度也不理想。尿液作为可以采用非侵入性方式收集的体液,其中含有大量来自于机体的蛋白质,是疾病标志物研究的良好来源。本研究采用定量蛋白质组学策略,将样本按照疾病进程分为健康对照(healthy individuals,HC)、结直肠癌无转移、结直肠癌淋巴转移(CRC with lymph node metastasis,CRC-LNM)以及结直肠癌肝转移(CRC with liver metastasis,CRC-LM)_4组,每组9例样本,用10标串联质谱标签(Tandem Mass Tags,TMT)标记,结合二维液相色谱串联质谱定量分析结直肠癌、结直肠癌淋巴转移、结直肠癌肝转移与健康个体之间蛋白含量的差异。结合质谱数据及实验室前期研究基础,选择肝细胞生长因子调节酪氨酸激酶底物(hepatocyte growth factor-regulated tyrosine kinase substrate,HGS)作为潜在标志物,通过敲降 HGS 观察结直肠癌细胞系表型的变化。研究发现,利用NC膜富集的方法可以从40mL的正常人和结直肠癌患者尿液中富集到300-800μg的蛋白,对分子量范围在20-130kDa的蛋白有较好的富集效果。定量蛋白质组学研究发现,四组中共有蛋白995个,差异蛋白富集在细胞凋亡及增殖通路;差异蛋白HGS与细胞迁移能力相关,可能是潜在的疾病标志物。胃癌的癌变过程是多因素长时间共同作用的结果。基因和环境会影响胃癌患病风险。已有的四项胃癌相关全基因组关联研究(genome-wide association study,GWAS)鉴定到 7 个单核苷酸多态性(single nucleotide polymorphism,SNP)位点,提示遗传因素在胃癌的发生发展过程中有一定的作用。同时,幽门螺杆菌(Helicobacterpylori,H.pylori)感染、吸烟和饮酒也是胃癌相关的重要环境因素。本研究目的是在胃癌中探索基因-环境相互作用,研究7个多态性位点与幽门螺杆菌感染、吸烟和饮酒在胃癌及重度肠上皮化生和不典型增生(severe intestinal metaplasia/dysplasia,severeIM/dysplasia)间的潜在交互作用。研究发现,PSCA rs2294008/rs2976392与幽门螺杆菌感染在胃癌风险上存在显著的相互作用。同时,PRKAAlrs13361707与幽门螺杆菌感染存在加法相互作用,SLC52A3 rs13042395与饮酒存在加法相互作用。此外,3个SNP位点,MUC1 rs4072037、ZBTB20 rs9841504和PRKAA1 rs13361707与癌前病变相关。提示多态性位点危险基因型可能与环境因素,特别是幽门螺杆菌感染和饮酒相互作用,增加胃癌风险。
[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.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735
，
本文编号：2350819
[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.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735
，
本文编号：2350819
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