环指蛋白180基因启动子甲基化在胃癌中的意义

发布时间：2018-03-27 23:21

  本文选题：胃癌　切入点：RNF180　出处：《天津医科大学》2016年硕士论文

【摘要】：目的检测胃癌组织中环指蛋白180(Ring finger protein 180,RNF180)DNA的核心功能启动子序列(-192/+126)内的43个CpG位点甲基化状态,分析胃癌组织RNF180抑癌基因启动子核心序列区43个CpG位点甲基化状态与胃癌患者预后、临床病理特征之间的关系,并分析可能存在的导致胃癌患者不良预后的关键性CpG位点,以期为胃癌患者的预后提供可靠标准。方法收集2003年4月~2007年12月就诊于天津医科大学肿瘤医院并施行胃癌根治术的400例胃癌患者临床病例资料,并从天津医科大学肿瘤医院病理组织库收集400例对应胃癌患者术后冰冻的胃癌组织,所有胃癌组织病理为胃腺癌,术前未行放疗、化疗、生物治疗等,提取胃癌组织DNA,利用亚硫酸氢盐基因组测序法(Bisulfite genomic sequencing,BGS)检测胃癌组织RNF180启动子核心序列区43个CpG位点甲基化状态。结果1.BGS测序法显示,400例患者抑癌基因RNF180核心功能启动子序列区发生甲基化的CpG位点数目呈0~43个不等,59例患者抑癌基因RNF180核心功能启动子序列区CpG位点无甲基化,341例患者呈现甲基化。2.运用Kaplan-Meier法分析出甲基化的CpG位点数的Cut-off值,51.25%(205/400)的患者甲基化CpG位点数≥8,48.75%(195/400)的胃癌患者甲基化CpG位点数≤7。经单因素分析,T分期(P=0.012)、N分期(P0.001)、淋巴结转移范围(P0.001)、Lauren分型(P=0.014)、发生甲基化的CpG位点数(P=0.002)、-116CpG位点的甲基化状态(P=0.041)、-80 CpG位点的甲基化状态(P=0.045)、+97 CpG位点的甲基化状态(P=0.021)、+102 CpG位点的甲基化状态(P=0.037)、联合(-116、-80、+97、+102)CpG位点的甲基化状态(P0.001)与胃癌患者术后的预后相关;多因素分析显示淋巴结分期(P0.001)、联合(-116、-80、+97、+102)CpG位点的甲基化状态(P=0.010)是胃癌术后的独立预后因素。3.RNF180 DNA的核心功能启动子序列区发生甲基化的CpG位点数与淋巴结转移位置显著相关:RNF180 DNA核心功能启动子区发生甲基化的CpG位点数≥8的胃癌患者的胃外淋巴结转移率(49.76%,102/205)明显高于发生甲基化的CpG位点数≤7的患者(34.87%,68/195)(P=0.010);同时RNF180 DNA核心功能启动子区发生甲基化的CpG位点数≥8的患者比发生甲基化的CpG位点数≤7的患者有更高的联合(-116、-80、+97、+102)CpG位点发生甲基化的率(91.70%v 13.33%,P0.001)。-80CpG位点发生甲基化的胃癌患者N3期淋巴结转移率(46.09%,59/128)比-80CpG位点未发生甲基化的患者N3期淋巴结转移率(35.66%,97/272)更高(P=0.024)。结论1.RNF180抑癌基因核心功能启动子区CpG位点的甲基化状态可以作为胃癌预后判断的重要分子指标。2.RNF180抑癌基因核心功能启动子区-116 CpG、-80 CpG、+97 CpG、+102CpG位点的甲基化在胃癌的形成发展中可能扮演着重要作用。
[Abstract]:Detection of gastric cancer tissue to ring finger protein 180 (Ring finger 180 protein, RNF180) the core function of DNA promoter sequence (-192/+126) of the 43 CpG sites methylation status, analysis of gastric cancer suppressor gene RNF180 promoter core sequence region 43 CpG sites methylation status and prognosis in patients with gastric cancer, the relationship between clinical pathology the characteristics, and analyze the possible cause critical CpG sites of poor prognosis of patients with gastric cancer, in order to provide a reliable standard for the prognosis of patients with gastric cancer. Methods from April 2003 December ~2007 in Medical University Of Tianjin hospital tumor hospital and 400 cases of patients with gastric cancer clinical data of gastric resection, and tumor pathology from hospital of Medical University Of Tianjin library collect 400 cases of gastric cancer patients with gastric cancer after operation corresponding to the frozen, all gastric cancer pathology of gastric cancer, radiotherapy, chemotherapy, biological therapy So, the extraction of DNA in gastric cancer tissues, using bisulfite genomic sequencing method (Bisulfite genomic sequencing, BGS) detection of gastric cancer tissue core sequence of RNF180 promoter region methylation status of 43 CpG sites. The results showed 1.BGS sequencing, 400 cases of tumor suppressor gene RNF180 in the core function number of CpG sites methylation of promoter sequences the area was 0~43 months, 59 cases of tumor suppressor gene RNF180 core function promoter sequence of CpG locus methylation free, 341 patients showed methylation analysis of.2. methylation of CpG sites in the Cut-off value by Kaplan-Meier method, 51.25% (205/400) patients with methylated CpG sites than 8,48.75% (195/400) the number of CpG methylation in gastric cancer patients less than 7. in the univariate analysis, T stage (P=0.012), N stage (P0.001), the extent of lymph node metastasis (P0.001), Lauren type (P=0.014), methylated CpG loci (P=0.002, -116Cp) The methylation status of G sites (P=0.041), the methylation status of -80 CpG loci (P=0.045), the methylation status of +97 CpG loci (P=0.021), the methylation status of +102 CpG loci (P=0.037), (-116, -80, and +97, +102) the methylation status of CpG loci (P0.001) associated with the prognosis of patients with gastric cancer; multivariate analysis showed that lymph node staging (P0.001), (-116, -80, and +97, +102) the methylation status of CpG locus (P=0.010) is the core function of independent prognostic factors of gastric cancer postoperative.3.RNF180 DNA promoter sequence region occurred methylated CpG sites and lymph node metastasis location: RNF180 DNA core function of stomach lymph CpG sites launched more than 8 gastric cancer patients with methylation sub area node metastasis rate (49.76%, 102/205) was significantly higher than that of methylated CpG sites is less than 7 of patients (34.87%, 68/195) and RNF180 DNA (P=0.010); core function Rev. The dynamic sub region of the CpG methylation sites of more than 8 of patients than the methylated CpG sites is less than 7 of patients with higher (-116, -80, +97, +102) CpG sites methylation rate (91.70%v 13.33%, P0.001) methylation loci of.-80CpG N3 in gastric cancer patients with stage lymph node metastasis rate (46.09%, 59/128) in patients with stage N3 than the -80CpG site without lymph node metastasis rate of methylation (35.66%, 97/272) higher (P=0.024). Conclusion the methylation status of 1.RNF180 tumor suppressor gene core promoter CpG sites can be important prognostic molecular marker of tumor suppressor gene.2.RNF180 the core function of the promoter region of -116 CpG, -80 CpG, +97 CpG, +102CpG locus in the formation of methylation may play an important role in the development of gastric cancer.

【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.2
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本文编号：1673720