SET8基因3’非翻译区的miR-502结合位点单核苷酸多态性与上皮性卵巢癌的相关研究

发布时间：2018-03-17 09:12

本文选题：上皮性卵巢癌　切入点：Micro　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:Micro RNA(mi RNA)是近年来发现的一类高度保守的小分子非编码RNA,长度约为22个核苷酸,人类基因组中有1/3以上基因是受其调控的。mi RNA通过与靶基因m RNA完全或不完全互补进而抑制靶m RNA编码的蛋白质的合成。越来越多的研究结果表明,mi RNA在许多生物过程中具有重要作用,如胚胎发育、细胞分化与增殖、细胞凋亡、肿瘤的发展和激素分泌等。单核苷酸多态性(Single Nucleotide Polymorphism,SNP)是指在基因组水平上由于单个核苷酸的改变引起的DNA序列的多态性,在人类基因组中分布广泛,是不同个体间最主要的遗传变异。mi RNA在靶基因上的结合位点的SNP可以改变靶基因的表达,从而影响患肿瘤的风险。SET8基因又称为SETD8,PR-SET7或KMT5a,编码组蛋白H4赖氨酸20单甲基转移酶,这种酶通过甲基化组蛋白进而调节基因的转录、细胞的生长周期及肿瘤的发生发展,SET8基因3'UTR的mi R-502结合区域内存在rs16917496多态位点,以往研究发现该位点多种肿瘤的发生和/或预后相关。本研究拟对SET8基因3’端非翻译区单核苷酸多态性rs16917496与上皮性卵巢癌患者的发病风险、临床特征及预后的相关性进行研究,为上皮性卵巢癌的早期诊断寻找新的分子靶点,并改善卵巢癌患者的预后。方法:1研究对象及随访选取100例2008年1月-2012年12月期间在河北医科大学第二医院进行妇科手术且术后组织学病理证实为上皮性卵巢癌的石蜡切片包埋组织切片,100例患者术前未接受任何治疗,详细记录患者相关临床资料(包括年龄、临床分期、残留癌灶大小等),通过电话随访患者术后的生存情况,随访截止时间2014年12月31号。另选取在河北医科大学第二医院体检健康的100例女性(除外与卵巢癌相关的疾病),获得知情同意后,详细记录体检资料,采集静脉血备用。2基因组DNA提取、扩增及测序:提取基因组DNA,DNA鉴定合格后,采用PCR技术扩增目的片段,大小为299bp,上游引物为:5’-CCTGGTCAG TGGTCAGCAAAT-3’,下游引物为:5’-CTGGGAAAC ACGCTCAAAA TC-3’。产物经琼脂糖凝胶电泳证实后进行双向重复测序。3统计学分析:采用SPSS21.0统计软件进行统计学分析,采用χ2检验或Fisher确切概率法比较计数资料。单因素分析使用Kaplan-Meier方法和Log-Rank方法,单因素分析中P值小于0.05的因素纳入COX风险回归模型进行多因素分析。P0.05被认为有统计学意义。结果:1在病例组与对照组之间,以基因型C/C为对照,与T/T基因型进行比较,差异无统计学意义(P0.05),与C/T、C/T+T/T基因型进行比较,差异有统计学意义(P㩳0.05),说明此位点与卵巢癌的发病风险相关,C/C基因型卵巢癌的发病风险较低。2病例组中rs16917496位点各基因型与各临床特征均无相关性,差异无统计学意义(P0.05)。3对上皮性卵巢癌患者进行生存分析,SET8基因的rs16917496多态位点C/C、C/T、T/T基因型患者的3年生存率分别为80%、74.1%、58.5%,尚不能认为此位点与上皮性卵巢癌患者的预后相关。4将单因素分析中有显著性差异的纳入COX模型进行多因素分析,结果显示,FIGO手术病理分期、残留癌大小与上皮性卵巢癌患者的死亡风险相关。结论:1 SET8基因3’端非翻译区的mi R-502结合位点单核苷酸多态性与上皮性卵巢癌的发病风险相关。2尚不能认为SET8基因3'端非翻译区mi R-502结合位点单核苷酸多态性与上皮性卵巢癌患者的预后相关。SET8基因的rs16917496位点基因型与上皮性卵巢癌患者的临床特征)均没有相关性。
[Abstract]:Objective: Micro RNA (MI RNA) is a recently discovered a highly conserved small molecule non encoding RNA, a length of about 22 nucleotides in the human genome contains more than 1/3.Mi RNA gene is under the control of the target gene m RNA complete or incomplete complementary protein synthesis and inhibition of target m RNA encoding. More and more studies show that MI RNA plays an important role in many biological processes, such as embryonic development, cell differentiation and proliferation, apoptosis, tumor development and hormone secretion. Single nucleotide polymorphisms (Single, Nucleotide Polymorphism, SNP) is a polymorphic DNA sequence caused by a single nucleotide change in the on genome level, widely distributed in the human genome,.Mi RNA is the main genetic variation among different individuals in the SNP binding sites of target gene can alter the expression of target genes, thus affecting the patient The risk of.SET8 gene is also known as SETD8, PR-SET7 or KMT5a, encoding histone H4 lysine 20 methylation transferase, the enzyme by methylation of histone regulates gene transcription, cell cycle and the occurrence and development of tumor cells, SET8 gene 3'UTR mi R-502 polymorphism in rs16917496 binding region of memory. Previous research found that the sites of tumor and / or prognosis. This research focuses on the risk of SET8 gene 3 'end untranslated region single nucleotide polymorphism and rs16917496 in patients with epithelial ovarian cancer, the correlation of clinical features and prognosis were studied to find new molecular targets for early diagnosis of epithelial ovarian cancer, and improve the prognosis of patients with ovarian cancer. Methods: 1 subjects and follow-up from 100 cases of January 2008 December -2012 during gynecological surgery and postoperative histology in the second hospital of Hebei Medical University Pathology of epithelial ovarian cancer paraffin embedded tissue sections, 100 patients did not receive any treatment, with clinical data of the patients were recorded (including age, clinical stage, residual tumor size), survival through telephone follow-up after surgery, follow-up deadline in December 31, 2014. Another selection in 100 women in the second hospital of Hebei Medical University medical health (except those associated with ovarian cancer disease), after obtaining informed consent, a detailed record of the physical examination data, extraction of venous blood sampling alternate.2 genomic DNA, amplification and sequencing of genomic DNA extraction, DNA: identification of qualified after amplification by PCR, size 299bp, upstream primer for the 5 '-CCTGGTCAG TGGTCAGCAAAT-3', as the downstream primer: 5 '-CTGGGAAAC ACGCTCAAAA TC-3'. The product was confirmed by agarose gel electrophoresis after double to repeat sequence of.3 statistics Analysis: using the statistical analysis software SPSS21.0, comparison of count data using 2 test or Fisher's exact probability method. Single factor analysis using Kaplan-Meier method and Log-Rank method, single factor analysis of P value is less than 0.05 of the factors in COX regression model for multivariate analysis.P0.05 was considered statistically significant. Results: 1 in between the case group and the control group, the genotype C/C were compared with T/T genotype, the difference was not statistically significant (P0.05), and C/T, compared with C/T+T/T genotype, the difference was statistically significant (P? 0.05), the risk of this locus and ovarian cancer, the risk of C/C genotype of ovary the low rs16917496 of.2 cancer patients locus genotypes and the clinical features were not correlated, no statistically significant difference (P0.05).3 in patients with epithelial ovarian cancer were survival analysis, SET8 gene rs16917 496 polymorphic loci C/C, C/T, the 3 year survival rate of patients with T/T genotype were 80%, 74.1%, 58.5%, still can not believe that the multi factor analysis, were incorporated into the COX model.4 this site and prognosis in patients with epithelial ovarian cancer. The single factor analysis showed that there was significant difference, FIGO staging the residual risk of death, and the size of the cancer patients with epithelial ovarian cancer. Conclusion: 1 SET8 gene 3 'untranslated region of the MI R-502 with the risk associated with single nucleotide polymorphisms of.2 and epithelial ovarian cancer is the SET8 gene 3' untranslated region of MI R-502 combined with clinical features, prognosis of.SET8 gene and single nucleotide polymorphism in patients with epithelial ovarian cancer. The rs16917496 genotype in patients with epithelial ovarian cancer) are not related.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.31

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