不同类型胰腺囊性肿瘤的基因组学及蛋白质糖基化的初步研究

发布时间：2018-07-11 10:56

本文选题：胰腺囊性肿瘤 + 黏液性囊腺瘤　；参考：《中国人民解放军医学院》2017年博士论文

【摘要】：目的:通过采用超声内镜下细针穿刺活检能够为胰腺囊性肿瘤的诊断提供直观的第一手资料,对肿瘤中发生基因突变的改变、蛋白质糖基化改变的进行初步的检测。方法:本实验主要分四部分进行:1.按病理类型将课题入组的胰腺囊性肿瘤患者分为高危组与低危组,选用多因素logistic回归分析低危组与高危组临床数据。2.对胰腺囊性肿瘤囊液进行DNA提纯,分析所选取的热点突变基因KRAS、GNAS、CTNNB1、BRAF在各类型中的突变情况。3.分析两大类型的胰腺囊性肿瘤,浆液性囊腺瘤与黏液性囊腺瘤囊液中蛋白质糖基化的变化情况:凝集素芯片技术比较浆液性/黏液性囊腺瘤囊液糖蛋白糖链谱。4.通过存在差异的凝集素富集,LC-MS/MS分析比较来自(N-linked)蛋白质肽段;完成蛋白质鉴定、通过生物信息学注释,及String蛋白相互作用图,寻找参与黏液性胰腺囊性肿瘤改变的通路和关键蛋白。结果:1.肿瘤的最长径≥2.5cm,主胰管≥5mm考虑为胰腺囊性肿瘤的危险因素。通过非参数秩和检验,囊液的CEA, CA19-9,CA72-4在低危组与高危组存在明显差异,结果显示脂肪酶≥1038.05ng/ml时,CEA≥10.33ng/ml时,诊断高危胰腺囊性肿瘤准确率为78.4%,有72.2%的敏感性和82.1%的特异性。2. 3例IPMN病例中其中1例KRAS基因12、13位点密码子的突变激活,Gly12CysGGTGTT,病理:胰腺导管内乳头状黏液性肿瘤(IPMN),部分上皮轻-中度不典型增生。10例MCN病例中其中1例CTNNB1突变热点为编码区:32/33/34/37密码子,34位点GGACGA,37位点TCTTCC,病理:胰腺(体尾部)黏液性囊腺瘤伴轻度不典型增生。基因突变检测可对胰腺囊性肿瘤的恶变起辅助诊断的意义。3. 6种凝集素(如STL、WGA、BPL、DBA、PTL-I、MAL-Ⅰ)识别糖链分别在胰腺囊性肿瘤中表达上调或下调。4.通过String蛋白相互作用分析,主要涉及代谢通路,PI3K-ATK 通路(YWHAZ,YWHAG,YWHAQ,LAMC1),处于交叉点核心的蛋白 YWHAZ,YWHAG,YWHAQ, ACTB, SPA5, S100A11均属于关键蛋白是参与黏液性胰腺囊性肿瘤改变的关键蛋白。结论:通过多个维度的对胰腺囊性肿瘤进行客观的诊断分析,研究胰腺囊性肿瘤相关糖蛋白糖链的变化,有助于寻找灵敏度和准确度更高的肿瘤标志物,进一步揭示胰腺囊性肿瘤进展的糖生物学机制并为研发新的治疗方法提供有价值信息。
[Abstract]:Objective: through the use of ultrasound-guided fine needle aspiration biopsy for the diagnosis of cystic tumor of the pancreas, the first hand data can be provided for the change of the gene mutation and the change of protein glycosylation in the tumor. Methods: this experiment is divided into four parts: 1. the cystic swelling of the pancreas in the group according to the pathological type The patients were divided into high risk group and low risk group. Multiple factor Logistic regression analysis was used to analyze the DNA purification of cystic fluid of pancreatic cystic tumor by.2. in low risk group and high risk group. Analysis of the selected hot mutation gene KRAS, GNAS, CTNNB1, BRAF in all types of mutations,.3. were divided into two types of pancreatic cystic tumors and serous cystadenoma. Changes in protein glycosylation in the cystic fluid of mucinous cystadenoma: the agglutinin chip technique compared the glycoprotein glycosyl chain.4. of the serous / mucinous cystadenoma with a differential agglutinin enrichment, LC-MS/MS analysis and comparison of the protein peptide from (N-linked), the identification of white matter, the bioinformatics annotation, and the String eggs. Results: the longest diameter of the 1. tumor was more than 2.5cm, and the main pancreatic duct more than 5mm was considered as a risk factor for pancreatic cystic tumors. The CEA, CA19-9, and CA72-4 of the fluid were significantly different in the low risk group and the high risk group by the nonparametric rank sum test. The results showed fat. When the enzyme was more than 1038.05ng/ml, when CEA was more than 10.33ng/ml, the accuracy of diagnosis of high risk pancreatic cystic tumors was 78.4%, with 72.2% sensitivity and 82.1% specific.2., of which 1 cases of KRAS gene 12,13 codon mutations were activated, Gly12CysGGTGTT, pathological: intraductal papillary mucinous tumor (IPMN), partial epithelial light to moderate. Among the cases of atypical hyperplasia, 1 cases of.10 MCN cases, CTNNB1 mutation hot spots are coded regions: 32/33/34/37 codon, 34 site GGACGA, 37 loci TCTTCC, pathology: mucinous cystadenoma of the pancreas (body tail) with mild atypical hyperplasia. Gene mutation detection can assist the diagnosis of pancreatic cystic neoplasia.3. 6 agglutinin (such as STL, WGA, BPL). DBA, PTL-I, MAL- I) recognize that the sugar chains are up-regulated and down regulated in cystic tumors of the pancreas, respectively, and.4. through String protein interaction analysis, mainly involved in the metabolic pathway, the PI3K-ATK pathway (YWHAZ, YWHAG, YWHAQ, LAMC1), and the protein YWHAZ at the core of the intersection. Conclusion: the objective diagnosis and analysis of cystic tumor of the pancreas by multiple dimensions and the study of the changes of glycoprotein glycate chain related to cystic tumor of the pancreas can help to find the tumor markers with higher sensitivity and accuracy, and further reveal the sugar biological mechanism of the progress of pancreatic cystic neoplasms. New methods of treatment provide valuable information.
【学位授予单位】：中国人民解放军医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R735.9

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