本文選題：胃癌　切入點：RNF180　出處：《天津醫(yī)科大學》2016年碩士論文

【摘要】：目的檢測胃癌組織中環(huán)指蛋白180(Ring finger protein 180,RNF180)DNA的核心功能啟動子序列(-192/+126)內的43個CpG位點甲基化狀態(tài),分析胃癌組織RNF180抑癌基因啟動子核心序列區(qū)43個CpG位點甲基化狀態(tài)與胃癌患者預后、臨床病理特征之間的關系,并分析可能存在的導致胃癌患者不良預后的關鍵性CpG位點,以期為胃癌患者的預后提供可靠標準。方法收集2003年4月~2007年12月就診于天津醫(yī)科大學腫瘤醫(yī)院并施行胃癌根治術的400例胃癌患者臨床病例資料,并從天津醫(yī)科大學腫瘤醫(yī)院病理組織庫收集400例對應胃癌患者術后冰凍的胃癌組織,所有胃癌組織病理為胃腺癌,術前未行放療、化療、生物治療等,提取胃癌組織DNA,利用亞硫酸氫鹽基因組測序法(Bisulfite genomic sequencing,BGS)檢測胃癌組織RNF180啟動子核心序列區(qū)43個CpG位點甲基化狀態(tài)。結果1.BGS測序法顯示,400例患者抑癌基因RNF180核心功能啟動子序列區(qū)發(fā)生甲基化的CpG位點數(shù)目呈0~43個不等,59例患者抑癌基因RNF180核心功能啟動子序列區(qū)CpG位點無甲基化,341例患者呈現(xiàn)甲基化。2.運用Kaplan-Meier法分析出甲基化的CpG位點數(shù)的Cut-off值,51.25%(205/400)的患者甲基化CpG位點數(shù)≥8,48.75%(195/400)的胃癌患者甲基化CpG位點數(shù)≤7。經單因素分析,T分期(P=0.012)、N分期(P0.001)、淋巴結轉移范圍(P0.001)、Lauren分型(P=0.014)、發(fā)生甲基化的CpG位點數(shù)(P=0.002)、-116CpG位點的甲基化狀態(tài)(P=0.041)、-80 CpG位點的甲基化狀態(tài)(P=0.045)、+97 CpG位點的甲基化狀態(tài)(P=0.021)、+102 CpG位點的甲基化狀態(tài)(P=0.037)、聯(lián)合(-116、-80、+97、+102)CpG位點的甲基化狀態(tài)(P0.001)與胃癌患者術后的預后相關;多因素分析顯示淋巴結分期(P0.001)、聯(lián)合(-116、-80、+97、+102)CpG位點的甲基化狀態(tài)(P=0.010)是胃癌術后的獨立預后因素。3.RNF180 DNA的核心功能啟動子序列區(qū)發(fā)生甲基化的CpG位點數(shù)與淋巴結轉移位置顯著相關:RNF180 DNA核心功能啟動子區(qū)發(fā)生甲基化的CpG位點數(shù)≥8的胃癌患者的胃外淋巴結轉移率(49.76%,102/205)明顯高于發(fā)生甲基化的CpG位點數(shù)≤7的患者(34.87%,68/195)(P=0.010);同時RNF180 DNA核心功能啟動子區(qū)發(fā)生甲基化的CpG位點數(shù)≥8的患者比發(fā)生甲基化的CpG位點數(shù)≤7的患者有更高的聯(lián)合(-116、-80、+97、+102)CpG位點發(fā)生甲基化的率(91.70%v 13.33%,P0.001)。-80CpG位點發(fā)生甲基化的胃癌患者N3期淋巴結轉移率(46.09%,59/128)比-80CpG位點未發(fā)生甲基化的患者N3期淋巴結轉移率(35.66%,97/272)更高(P=0.024)。結論1.RNF180抑癌基因核心功能啟動子區(qū)CpG位點的甲基化狀態(tài)可以作為胃癌預后判斷的重要分子指標。2.RNF180抑癌基因核心功能啟動子區(qū)-116 CpG、-80 CpG、+97 CpG、+102CpG位點的甲基化在胃癌的形成發(fā)展中可能扮演著重要作用。
[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.

【學位授予單位】：天津醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R735.2
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本文編號：1673720