基质金属蛋白酶MMPs及亚甲基四氢叶酸还原酶MTHFR基因多态性与宫颈癌的相关性研究

发布时间：2018-03-11 21:41

本文选题：基质金属蛋白酶　切入点：亚甲基四氢叶酸还原酶　出处：《山东大学》2016年博士论文　论文类型：学位论文

【摘要】：背景宫颈癌是女性生殖道最常见的恶性肿瘤,近年来发病有年轻化的趋势。目前认为高危型人乳头瘤病毒(HPV)的持续性感染,是引起宫颈癌的基本原因。然而在众多HPV感染者中,仅有极少数人最终发展成宫颈癌。人群中HPV的高感染率与宫颈癌实际发病率之问存在明显的差异,提示在宫颈癌的发生过程中,遗传易感因素起到不容忽视的作用。宫颈癌实际上是外源性感染因素和内源性易感因素共同作用的结果,目前宫颈癌发病的遗传易感因素包括基因单核苷酸多态性(SNPs)已经成为研究的一个方向。基质金属蛋白酶(MMPs)是一类活性依赖于锌离子或钙离子的蛋白水解酶,可降解细胞外基质(ECM),在肿瘤的浸润、迁移及血管生成等过程中均发挥着重要作用；目前关于MMPs家族基因多态性与宫颈癌的相关性研究报道不多。亚甲基四氢叶酸还原酶(MTHFR)是叶酸代谢过程中的关键酶,其基因多态性与多种恶性肿瘤的发生密切相关。有报道,亚甲基四氢叶酸还原酶MTHFR的基因多态性与子宫颈癌的发生相关,然而国内外研究结论并不一致。本研究通过对宫颈癌患者与对照人群中MMPs及MTHFR基因多态性的对比,探讨基质金属蛋白酶(MMPs)及亚甲基四氢叶酸还原酶(MTHFR)基因多态性与宫颈癌遗传易感性及临床病癌特征的关系。目的采用病例对照研究方法,探讨基质金属蛋白酶MMP2-1306 C/T、 MMP3-11715A/6A、MMP7-181A/G、MMP9-1562C/T以及亚甲基四氢叶酸还原酶MTHFR 677C/T、MTHFR1298A/C基因多态性与宫颈癌易感性及临床病理特征的关系,寻找宫颈癌易感因素,为宫颈癌早期诊断、针对性预防提供理论依据。方法研究对象为2012年1月-2016年1月在我院就诊的宫颈癌患者,选取同期因其他良性疾病就诊者为对照组。病例组及对照组所有入选研究对象均于清晨空腹状态取肘静脉血3ml,置于一次性抗凝管中混匀,入-40℃冰箱内保存,以提取全血基因组DNA。采用聚合酶链反应(PCR)和限制性片段长度多态(RFLP)技术检测MMP2-1306 C/T、MMP3-11715A/6A、MMP7-181A/G、MMP9-1562C/T、 MTHFR 677C/T、MTHFR1298A/C多态性位点的基因型。留取宫颈癌(Ⅰ-Ⅱa期)手术治疗患者的病理标本,取材后立即放入液氮罐中,转移至-80℃保存备用,以提取DNA,通过PCR-RFLP技术对多态性位点的基因型进行检测。因良性病变切除子宫患者的宫颈标本作为对照。采用SPSS19.0统计软件对所有数据进行整理分析,计数资料采用卡方检验；病例组及对照组基因型频率分布进行Hardy-weinberg平衡检验；Logistic回归法进行相关性分析。P0.05差异具有统计意义。结果第一部分MMPs及MTHFR基因多态性与宫颈癌的相关性1、MMP2-1306 C/T、MMP3-11715A/6A、MMP7-181A/G、MMP9-1562C/T、 MTHFR 677C/T和MTHFR1298A/C的基因型频率分布符合Hardy-Weinberg平衡,研究对象具有代表性。2、MMP2-1306 C/T、MMP7-181A/G及MTHFR 677C/T的基因多态性与宫颈癌有显著相关性(p0.05)。3、MMP3-11715A/6A、MMP9-1562C/T及MTHFR1298A/C的基因多态性与宫颈癌无显著相关性(p0.05)。第二部分MMPs及MTHFR基因多态性与宫颈癌临床病理特征的相关性1、MMP2-1306 C/T、MMP7-181A/G及MTHFR 677C/T的基因多态性与宫颈癌分期有显著相关性(p0.05)。2、MMP2-1306 C/T、MMP7-181A/G及MTHFR 677C/T的基因多态性与宫颈癌患者的淋巴结转移没有显著相关性(p0.05)。3、MMP2-1306 C/T、MMP7-181A/G及MTHFR 677C/T的基因多态性与宫颈癌的病理组织类型无显著相关性(p0.05)。4、宫颈癌患者中MTHFR 677C/T的基因多态分布与MMP2的基因多态分布具有显著相关性(p0.05),而与MMP7的基因多态分布无显著相关性(p0.05)。结论MTHFR 677C/T、MMP2-1306 C/T和MMP7-181A/G的基因多态性与宫颈癌的发生和分期有显著相关性,临床可根据患者不同基因型分群进行特异性调整干预方案。
[Abstract]:Background: cervical cancer is the most common female reproductive tract cancer incidence in recent years, the trend of younger. The high-risk human papillomavirus (HPV) persistent infection, is the basic causes of cervical cancer. However, in many HPV infection, only a handful of people eventually develop into cervical cancer. There are obvious differences among the high HPV infection rate and the actual incidence of cervical cancer asked, prompt in the process of cervical carcinogenesis in genetic susceptibility plays a role can not be ignored. Cervical cancer is actually exogenous and endogenous infection factors predisposing factors common result, the heredity of cervical cancer the incidence of predisposing factors including single nucleotide polymorphism (SNPs) has become a research direction. The matrix metalloproteinase (MMPs) is a kind of proteolytic activity is dependent on zinc or calcium ions, degradation of extracellular matrix (ECM), in tumor invasion, migration and angiogenesis plays an important role; there are few reports about the correlation between MMPs gene polymorphism and familial cervical cancer. The methylenetetrahydrofolate reductase (MTHFR) is a key enzyme in folate metabolism, its gene polymorphism and a variety of malignant tumors closely related. There are reports, genetic polymorphisms of methylenetetrahydrofolate reductase MTHFR and the occurrence of cervical cancer, but the findings of research at home and abroad are not consistent. This study of cervical cancer patients with contrast MMPs and MTHFR gene polymorphisms in the crowd, to investigate the expression of matrix metalloproteinase (MMPs) and methylenetetrahydrofolate reductase (MTHFR) the relationship between gene polymorphism and susceptibility to cervical cancer and the clinical features of the disease cancer. Objective to case-control study were to investigate the expression of matrix metalloproteinase MMP2-1306 C/T, MM P3-11715A/6A, MMP7-181A/G, MMP9-1562C/T and methylenetetrahydrofolate reductase MTHFR 677C/T, the relationship between MTHFR1298A/C gene polymorphism and the susceptibility of cervical cancer and the clinical pathological features of cervical cancer, find the predisposing factors for early diagnosis of cervical cancer, targeted prevention and provide a theoretical basis. The research object for the January 2012 -2016 year in January in our hospital in patients with cervical cancer and the same time for other benign disease patients as the control group. The case group and control group all subjects were in the early morning fasting state of umbilical venous blood 3ml, a one-time anti coagulation tube in mixing, into the refrigerator to save -40 C, genomic DNA. extraction by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) detection of MMP2-1306 C/T MMP3-11715A/6A, MMP7-181A/G, MMP9-1562C/T, MTHFR, 677C/T, MTHFR1298A/C genotype polymorphism. Stay The cervical cancer (stage I-II a) pathological specimens of patients with surgical treatment, were immediately placed in liquid nitrogen tank, transferred to the -80 stored at standby, to extract the DNA gene were detected by PCR-RFLP technology on polymorphism. Due to benign lesion of uterine cervical specimens were chosen as control. SPSS19.0 the statistical software analysis of all data, count data using chi square test; case group and control Hardy-weinberg group balance test genotype frequency distribution; correlation analysis.P0.05 the difference has statistical significance of Logistic regression method. The correlation between the first part of the MMPs and MTHFR gene polymorphism and cervical cancer 1, MMP2-1306 C/T, MMP3-11715A/6A. MMP7-181A/G, MMP9-1562C/T, 677C/T and MTHFR genotype frequencies of MTHFR1298A/C with the Hardy-Weinberg equilibrium, the research object is representative of.2, MMP2-1306 C/T, MMP7 There was a significant correlation between -181A/G and MTHFR 677C/T gene polymorphism and cervical cancer (P0.05).3, MMP3-11715A/6A, MMP9-1562C/T and there is no significant correlation between MTHFR1298A/C gene polymorphism and cervical cancer (P0.05). The correlation between the second parts of MMPs and MTHFR gene polymorphism and clinicopathological features of cervical cancer was 1 MMP2-1306, C/T, MMP7-181A/G and MTHFR 677C/T gene polymorphism and cervical cancer staging had significant correlation (P0.05).2, MMP2-1306 C/T, MMP7-181A/G and MTHFR with 677C/T gene polymorphism and cervical cancer lymph node metastasis has no significant correlation (P0.05).3, MMP2-1306 C/T, there is no significant correlation between pathological types of MMP7-181A/G and MTHFR gene polymorphism and cervical cancer (677C/T P0.05).4 gene polymorphism in patients with cervical cancer in MTHFR 677C/T and MMP2 distribution has significant correlation (P0.05), and the distribution and MMP7 gene polymorphism There was no significant correlation (P0.05). Conclusion MTHFR 677C/T, MMP2-1306 C/T and MMP7-181A/G gene polymorphisms are significantly correlated with the occurrence and staging of cervical cancer. According to the different genotypes of patients, clinical intervention can be adjusted.

【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R737.33

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