microRNA生物合成基因多态性与鼻咽癌和乙型肝炎病毒相关肝癌的遗传关联研究

发布时间：2018-04-02 04:36

本文选题：肝癌　切入点：鼻咽癌　出处：《安徽医科大学》2012年硕士论文

【摘要】：microRNA（简称miRNA）生物合成基因是指参与miRNA基因转录成pri-miRNA、pri-miRNA被剪切为pre-miRNA，以及pre-miRNA被运输到细胞质内形成成熟miRNA的一类基因。单核苷酸多态性（single nucleotide polymorphisms,SNPs）是指基因组水平上由单个核苷酸的改变引起的DNA序列多态性，包括单个碱基的转换、颠换、插入和缺失等形式。SNPs与个体的表型差异、不同个体对复杂疾病的易感性和临床转归的差异，以及药物敏感性的差异等显著相关。miRNA生物合成基因中的SNPs可以影响该类基因的生物学功能，进而影响miRNA对靶基因的调控。已有多项研究显示miRNA生物合成基因的多态性与包括肿瘤在内的多种疾病的发生发展密切相关。但是，迄今尚未见miRNA生物合成通路基因中的多态性与肝癌或鼻咽癌的遗传关联研究的报道。因此，我们系统开展了miRNA生物合成通路基因多态性与中国人群肝癌的易感性，以及与鼻咽癌的遗传易感性和严重程度之间的遗传关联研究。主要结果如下：一、miRNA生物合成通路基因多态性与鼻咽癌的易感性和严重程度的相关性研究我们在miRNA生物合成通路基因中选择了97个SNPs位点，分别在招募自广西的鼻咽癌病例-对照人群（包括855个病例和1036个对照）、招募自广东的鼻咽癌病例-对照（997个病例和972个对照）和招募自广西的核心家系人群（包括222个核心家系）中进行了遗传关联分析。结果发现，AGO2基因内含子区的SNP位点rs3928672与鼻咽癌的易感性和淋巴结转移显著相关。与携带GG基因型的个体相比，携带GA和AA基因型的个体发生鼻咽癌的风险显著增加（OR=1.20，95%CI=1.05-1.36，P=0.006）；同时，携带GA和AA基因型的个体发生鼻咽癌淋巴结转移的风险也显著增加（OR=1.97，95%CI=1.37-2.83，P=0.003）。连锁不平衡分析显示，，没有与rs3928672呈强连锁不平衡的SNPs。进一步根据性别、年龄、饮酒和吸烟状态、以及居住地进行分层分析显示，rs3928672与吸烟因素存在交互作用效应，而与其它混杂因素无显著的交互作用。rs3928672位于AGO2基因内部，通过生物信息学方法进行功能预测显示，rs3928672可能影响AGO2基因的转录水平。对37例鼻咽癌组织和18例非鼻咽癌个体的鼻咽部组织的免疫组化分析显示，AGO2在鼻咽癌组织中异常高表达，并且，随着鼻咽癌N分期的增加，AGO2的表达有增高的趋势。在人鼻咽癌细胞系CNE2Z中抑制AGO2表达，发现能显著降低CNE2Z的迁移能力。抑制AGO2表达前后的miRNA和mRNA表达谱研究显示，AGO2的扰动能导致大量mRNA和miRNA的表达发生改变，并且这些发生改变的基因显著富集于“RAS”、“ERK”、“MAPK”和“Invasion”等与肿瘤发生发展密切相关的信号通路。上述这些功能学研究结果均支持AGO2基因多态性与鼻咽癌的发生风险和淋巴结转移的遗传学关联结果，进一步证明AGO2在鼻咽癌的发生发展中发挥重要作用。二、miRNA生物合成通路基因多态性与乙型肝炎病毒相关肝癌易感性的研究我们在miRNA生物合成通路基因中选择了178个SNPs位点，在招募自广西的HBV相关肝癌病例-对照人群（包括358个病例和373个对照）中进行了遗传关联分析。结果发现，有3个SNPs位点与HBV相关肝癌的遗传易感性显著相关，它们分别是PACT rs3752689（OR=1.42，95%CI=1.11-1.81，P=0.006）、IPO8rs6487924（OR=1.60，95%CI=1.09-2.37，P=0.002）和AGO2rs938726（OR=2.26，95%CI=1.41-3.61，P=0.0003）。进一步在招募自广东的HBV相关肝癌病例-对照人群（包括751个病例和509个对照）中对上述关联结果进行了重复验证研究，发现rs6487924与HBV相关肝癌的遗传易感性仍然显著相关（OR=1.49，95%CI=1.08-2.07，P=0.010）。这些结果提示，miRNA生物合成相关基因多态性可能在HBV相关肝癌的易感性中起重要作用。
[Abstract]:MicroRNA (miRNA) biosynthesis gene is involved in transcription of miRNA gene into pri-miRNA, pri-miRNA was cut into pre-miRNA, a class of genes and pre-miRNA is transported to the cytoplasm of mature miRNA. Single nucleotide polymorphism (single nucleotide polymorphisms, SNPs) is a DNA sequence polymorphism in the genome level caused by a single nucleotide change. Conversion, including single nucleotide transversion, insertion and deletion phenotypes.SNPs and individual differences in outcomes, different individuals to complex disease susceptibility and clinical, and drug sensitivity significantly related to.MiRNA biosynthesis genes in SNPs can affect the biological function of the gene, and the regulatory effects of miRNA on target gene. Several studies have shown that the miRNA biosynthetic gene polymorphism with a variety of diseases including tumor development. Cut. However, so far there is no genetic association study miRNA biogenesis genes polymorphism with HCC or nasopharyngeal carcinoma reported. Therefore, we carried out miRNA biosynthesis pathway susceptibility gene polymorphism and China liver cancer, and the genetic association study between genetic susceptibility and severity of nasopharyngeal carcinoma and the main results are as follows:
The correlation between the polymorphism of miRNA biosynthetic pathway gene and the susceptibility and severity of nasopharyngeal carcinoma
We chose 97 SNPs sites in the miRNA biosynthesis pathway genes, respectively, in the case of nasopharyngeal carcinoma recruited from Guangxi group (including 855 cases and 1036 controls), recruitment control since nasopharyngeal carcinoma - Guangdong (997 cases and 972 controls) and were recruited from core pedigrees in Guangxi (including 222 core pedigrees) conducted genetic association analysis. The results showed that SNP was significantly associated with nasopharyngeal carcinoma AGO2 gene rs3928672 locus of intron in susceptibility and lymph node metastasis. Compared with GG genotype, significantly increased the risk of carrying GA and AA genotype had nasopharyngeal carcinoma (OR=1.20,95%CI=1.05-1.36, P=0.006); at the same time, carrying GA and AA genotype risk of lymph node metastasis in nasopharyngeal carcinoma was also significantly increased (OR=1.97,95%CI=1.37-2.83, P=0.003). Linkage disequilibrium analysis showed that there was rs3928672 Strong linkage disequilibrium SNPs. according to gender, age, smoking and drinking status, and residence were stratified analysis showed that the rs3928672 and smoking factor interaction effects, and other confounding factors did not interact significantly at.Rs3928672 in AGO2 gene, function prediction showed by bioinformatics method, the transcription level of rs3928672 may effect of AGO2 gene.
The immune group of 37 cases of nasopharyngeal carcinoma tissues and 18 cases of nasopharyngeal carcinoma of the nasopharynx tissue of individual analysis showed that AGO2 in nasopharyngeal carcinoma abnormal expression, and, with the increase of N staging for nasopharyngeal carcinoma, the expression of AGO2 increased. The inhibition of AGO2 expression in human nasopharyngeal carcinoma cell line CNE2Z, and found a significant reduction in migration the ability of CNE2Z to inhibit the expression of AGO2 and miRNA. The expression of mRNA and AGO2 spectral studies show that disturbance can lead to a high expression of mRNA and miRNA changed, and these changes were significantly enriched in "RAS", "ERK", "MAPK" and "Invasion" associated with signal transduction the risk of occurrence of genetic association. The tumor and lymph node metastasis. These results support the functional studies of AGO2 gene polymorphism and nasopharyngeal carcinoma, further evidence of AGO2 in the development of nasopharyngeal cancer. Play an important role.
Two, miRNA biosynthesis pathway gene polymorphism and susceptibility to HBV associated hepatocellular carcinoma
We chose 178 SNPs sites in the miRNA biosynthesis pathway genes, in the recruitment of HBV related HCC cases from Guangxi -- control group (including 358 cases and 373 controls) were carried out in genetic association analysis. The results showed that there were significantly related to genetic susceptibility of 3 SNPs loci and HBV related hepatocellular carcinoma, respectively PACT rs3752689 (OR=1.42,95%CI=1.11-1.81, P=0.006), IPO8rs6487924 (OR=1.60,95%CI=1.09-2.37, P=0.002) and AGO2rs938726 (OR=2.26,95%CI=1.41-3.61, P=0.0003). Further in HBV related HCC cases were recruited from Guangdong control group (including 751 cases and 509 controls) of the correlation results were repeated validation studies, found that genetic susceptibility to rs6487924 and HBV related hepatocellular carcinoma the remained significant (OR=1.49,95%CI=1.08-2.07, P=0.010). These results suggest that miRNA biosynthesis related gene polymorphism in HBV The susceptibility to liver cancer plays an important role.

【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R735.7;R739.63

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