MicroRNA单核苷酸多态性与肝细胞癌易感性及MiR-196a2rs11614913与肝癌预后相关性研究

发布时间：2019-04-08 20:32

【摘要】：第一部分Micro RNA单核苷酸多态性与肝细胞癌易感性目的:本文拟采用自然人群为基础病例对照研究,旨在揭示位于mi RNA编码区和作用靶点的基因多态性与肝癌发病风险可能的关系。综合分析并揭示环境因素、遗传因素与mi RNA的交互作用对肝癌发病风险的影响。并根据肝癌遗传易感性的分析结果,探讨mi RNA不同基因型在易感患者血肿的的表达特性,为临床对于肝癌预防性诊断提供参考。方法:随机选取内蒙古医科大学附属医院2013年9月-2016年2月266例肝细胞癌患者作为实验组,266例来我院体检的正常无癌病人为对照组。统计实验组的性别、年龄、高血压、糖尿病、吸烟、饮酒、病毒感染、肿瘤大小和肿瘤阶段。收集两组患者静脉血5m L样品,通过聚合酶链式反应限制性片段多态性试剂盒,进行测量;通过聚合酶链反应对DNA扩增。mi R-146a2 rs2910164,mi R-196a2rs11614913,mi R-149 rs2292832和mi R-499 rs3746444的基因型,通过RT-PCR法进行扩增,扩增后全部测序。通过病例对照研究,综合评估各个基因多态性与肝癌遗传易感性的关系,并计算相应的比值比和95%可信区间。结果:(1)本研究发现高血压、糖尿病、吸烟与患肝癌风险差异无统计学意义。饮酒和病毒感染与肝癌发病风险显著相关。(2)对位于mi RNA编码区的基因多态性与肝癌发病风险关系的分析结果表明,mi R-196(TT基因型和T等位基因基因)编码区的基因多态性位点与肝癌的发病风险显著增加,分别增加了2.29倍和1.60倍(95%CI=1.11-2.32),而mi R-146、mi R-149和mi R-499的多态性与肝癌危险性不相关。(3)HBV感染者mi R-196a2(CT+TT基因型)可能是增加肝细胞癌的危险性的原因,性别、年龄和酒精中毒mi R-196的多态性和肝细胞癌危险性之间可能没有相关性。结论:(1)mi R-196a2 rs11614913的TT基因型和T等位基因与肝细胞癌危险性增加有显著相关性,该两种基因型可能成为肝细胞癌的诊断的标记。(2)HBV感染显著影响mi R-196a2 rs11614913的多态性与肝细胞癌危险性之间的关系。(3)mi R-196a2 rs11614913的TT基因型和T等位基因可能是肝细胞癌患病危险性增加的标记。(4)mi R-196a2 rs11614913的多态性可能是影响肝细胞癌的发展的因素之一,特别是感染HBV的患者。第二部分Mi R-196a2rs11614913多态性与肝癌预后关系的研究目的:由实验一可以发现Mi R-196a的多态性与肝癌的发病可能相关,本部分的研究就Mi R-196a基因多态性与原发性肝癌预后相关指标之间的相关性进行分析,旨在为指导肝细胞癌化疗药物的治疗和评估患者的预后及生存时间提供可能的参考。方法:随机选取2013年9月-2016年2月123例肝细胞癌内蒙古医科大学附属医院肝细胞癌患者作为实验组。研究对象采用RT-PCR方法检测其Mi R-196a2rs11614913基因多态性;采用基因计数方法检验对象等位基因频率和基因型频率。药物治疗组:肝癌术后手术后1个月选用FOLFOX(奥沙利铂+亚叶酸钙+氟尿嘧啶)联合治疗,全部患者均愿意在手术后接受化疗,术前后基线资料均从临床病例中获得,病人出院后每隔四周电话随访或者门诊随访,直到患者死亡或本次研究结束。对比基线资料,观察Mi R-196a2rs11614913基因多态性类型,对比Mi R-196a2rs11614913基因多态性检测情况。结果:选取123例肝癌患者中,90例(73.2%)肿瘤直径小于5cm,33例(26.8%)肿瘤直径大于5cm,47例(38.2%)显示为肿瘤TNM分期中的II期,76例(61.8%)显示为III-IV期。Mi R-196a基因型在化疗药物敏感组和不敏感组间差异有统计学意义(χ2=11.91,P=0.001)。与Mi R-196a功能基因型相比较,Mi R-196a缺陷基因型患者生存时间较长,差异具有统计学意义。Mi R-196a基因缺陷型显著降低了肝癌的病死风险,HR(95%CI)为0.49(0.25-0.96)。结论:(1)Mi R-196a2rs11614913的多态性与患者对化疗药物敏感性和生存时间相关。而与Mi R-196a2rs11614913功能型相比较,Mi R-196a2rs11614913的缺陷型基因型的肝癌患者对化疗药物更敏感。(2)Mi R-196a2rs11614913基因型患者基因型生存期短。(3)Mi R-196a2rs11614913多态性阳性的患者可以为患者制定个性化的化疗方案提供参考。
[Abstract]:Objective: To study the relationship between the polymorphism of the first partial micro-RNA and the susceptibility of hepatocellular carcinoma: a case-control study based on the natural population is proposed to reveal the possible relationship between the gene polymorphism of the mi-RNA coding region and the target of action and the risk of liver cancer. The effects of environmental factors, genetic factors and the interaction of mi-RNA on the risk of liver cancer were analyzed. According to the analysis result of the genetic susceptibility of the liver cancer, the expression characteristics of the mi-RNA different genotypes in the susceptible patients haematoma are discussed, and the reference for the preventive diagnosis of the liver cancer is provided for clinic. Methods: From September 2013 to February 2016,266 cases of hepatocellular carcinoma were randomly selected as experimental group and 266 cases of normal non-cancer patients in our hospital as the control group. The sex, age, hypertension, diabetes, smoking, drinking, viral infection, tumor size and tumor stage of the experimental group were counted. The 5-m L samples from the venous blood of the two groups were collected and measured by the restriction fragment polymorphism kit of the polymerase chain reaction; and the DNA was amplified by the polymerase chain reaction. The genotype of mi R-146a2 rs2910164, mi R-196a2rs11614913, mi R-149rs2292832 and mi R-499 rs374644 was amplified by RT-PCR and all sequenced after amplification. The relationship between each gene polymorphism and the genetic susceptibility of the liver cancer was assessed by a case-control study, and the corresponding ratio ratio and the 95% confidence interval were calculated. Results: (1) There was no significant difference in the risk of hypertension, diabetes, smoking and the risk of liver cancer. Alcohol consumption and viral infection were associated with the risk of liver cancer. (2) The results of the analysis of the relationship between the gene polymorphism and the risk of liver cancer in the mi-RNA coding region show that the genetic polymorphism site of the mi R-196 (TT genotype and T allele gene) coding region is increased by 2.29 times and 1.60 times (95% CI = 1.11-2.32), and the mi R-146, respectively. The polymorphisms of mi R-149 and mi R-499 are not associated with the risk of liver cancer. (3) The mi R-196a2 (CT + TT genotype) of HBV-infected persons may be the cause of increasing the risk of hepatocellular carcinoma, and there may be no correlation between the polymorphism of the mi R-196 and the risk of hepatocellular carcinoma. Conclusion: (1) The TT genotype and T allele of mi R-196 a2 rs11614913 are significantly associated with the increase of the risk of hepatocellular carcinoma, which may be a marker of the diagnosis of hepatocellular carcinoma. (2) The relationship between the polymorphism of mi R-196 a2 rs11614913 and the risk of hepatocellular carcinoma was significantly affected by HBV infection. (3) The TT genotype and T allele of mi R-196 a2 rs11614913 may be an increased risk for hepatocellular carcinoma. (4) The polymorphism of mi R-196 a2 rs11614913 may be one of the factors that affect the development of hepatocellular carcinoma, especially those with HBV. The study of the relationship between the polymorphism of the second part Mi R-196a2rs11614913 and the prognosis of the liver cancer is to study the correlation between the polymorphism of the Mi R-196a and the prognosis of the primary liver cancer. It is intended to provide a possible reference for the treatment and evaluation of the prognosis and survival time of a patient with a chemotherapeutic agent for hepatocellular carcinoma. Methods: From September 2013 to February 2016,123 patients with hepatocellular carcinoma from Inner Mongolia Medical University of Inner Mongolia were selected as experimental group. The gene polymorphism of Mi R-196 a2rs11614913 was detected by RT-PCR. The treatment group: FOLFOX (oxaliplatin + calcium folinate + fluorouramide) was used in the first month after the operation of the liver cancer. All the patients were willing to receive the chemotherapy after the operation, and the baseline data before and after the operation were obtained from the clinical cases. The patient was followed up for follow-up or out-of-patient follow-up every four weeks after the discharge of the patient until the patient died or the study was completed. The genetic polymorphism of the Mi R-196a2rs11614913 gene was observed compared with the baseline data, and the genetic polymorphism of the Mi R-196a2rs11614913 gene was compared. Results: Of the 123 patients with liver cancer,90 (73.2%) of the tumors had a tumor diameter of less than 5 cm,33 (26.8%) of the tumors had a tumor diameter of more than 5 cm, and 47 (38.2%) were shown as phase II in the TNM stage of the tumor and 76 (61.8%) were shown as phase III-IV. The difference of Mi R-196a genotype between sensitive and non-sensitive groups was statistically significant (Sup2 = 11.91, P = 0.001). Compared with the functional genotype of the Mi R-196a, the survival time of the Mi R-196a deficient genotype was longer and the difference was of statistical significance. The genetic defect of Mi R-196a significantly reduced the risk of dying of liver cancer, and HR (95% CI) was 0.49 (0.25-0.96). Conclusion: (1) The polymorphism of Mi R-196a2rs11614913 is related to the sensitivity and survival time of chemotherapy drugs. In contrast to the functional phase of the Mi R-196 a2rs11614913, the patients with a defective genotype of Mi R-196a2rs11614913 were more sensitive to chemotherapy. (2) The genotype of the Mi R-196a2rs11614913 genotype was short. (3) Patients with the Mi R-196a2rs11614913 polymorphism can provide a reference for the patient to develop a personalized chemotherapy regimen.
【学位授予单位】：重庆医科大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R735.7

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