HIF-1α基因rs2057482SNP位点的群体调查、脑梗死相关性及法医学意义研究

发布时间：2018-06-25 12:24

本文选题：HIF-1α基因 + 单核苷酸多态性　；参考：《中国医科大学》2009年硕士论文

【摘要】： 前言缺氧诱导因子1(hypoxia-inducible factor-1,HIF-1)是目前发现的唯一特异性缺氧状态下发挥活性的转录因子,于1992年首先由Semenza等在低氧的肝细胞癌细胞株Hep3B细胞的核提取物中发现。HIF-1广泛存在于哺乳动物和人体内,是由α和β两个亚单位组成的一种异源二聚体。其中HIF-1α是唯一的氧调节亚单位,只有在低氧或缺氧情况下才表现其生物学活性,它主要通过与靶基因的缺氧反应元件(hypoxia response element,HRE)上的HIF-1结合位点(5'-TACGTG CT-3')相结合,从而激活靶基因的转录,发挥一系列的生物学功能,其中包括参与血管生成、能量代谢、红细胞生成、细胞增殖和存活、血管重塑以及血管舒缩反应等。编码HIF-1α的基因位于14号染色体q21-24区,该区域单核苷酸多态性十分丰富,截止目前,NCBI已公布了238个SNPs位点,其中研究较多的是编码区的SNPs位点。研究发现此区域的单核苷酸多态性可以影响蛋白质表达和功能,国外研究多集中在HIF1α基因遗传多态性与各种肿瘤(如泌尿、消化、生殖等系统肿瘤)关系研究,也有报道与糖尿病、心肌缺血梗死以及非创伤性股骨头坏死等疾病密切相关。而HIF-1α基因在中国辽宁汉族、广西壮族人群的分布及其法医学应用价值国内外尚无报道。另外,研究发现HIF-1α与脑缺血的关系密切,在脑缺血的保护性反应中起重要作用,目前国内外尚无HIF-1α的基因多态性与脑缺血缺氧性疾病相关性的研究报道。本研究选择HIF-1α基因rs2057482 SNP位点,对辽宁汉族和广西壮族人群rs2057482位点单核苷酸多态性进行基因分布调查,并对该基因与脑梗死疾病的关系进行了探讨,旨在为人类遗传学、临床医学以及法医学等领域提供有价值的参考数据。材料与方法 1、259份辽宁汉族、191份广西壮族无血缘关系健康献血者及159份脑梗死患者的血液样本。10个2代3口家系共30份DNA样品。 2、应用双向等位基因特异性扩增(Bi-ASA)技术调查HIF-1α基因rs2057482SNP位点在辽宁汉族、广西壮族健康人群以及脑梗死患者中的分布状况。并进行家系调查。 3、直接计数法计算基因型及等位基因频率;Hardy-Weinberg平衡检验所研究群体是否具有代表性;应用SPSS15.0统计软件进行不同人群基因分布差异以及对照组与病例组组间分析。计算法医学参数:杂合度(H)、多态性信息含量(PIC)、个人识别率(DP)和非父排除率(EPP)。结果在三个群体中,均检测出HIF-1α基因rs2057482位点的C、T两个等位基因,及CC、CT、TT三种基因型。在辽宁汉族群体中,C、T等位基因频率分别为0.8301、0.1699;CC、CT、TT基因型频率分别为0.6873、0.2857、0.0270。广西壮族群体中C、T基因频率分别为0.9031,0.0969;CC、CT、TT的频率分别为0.8272、0.1518、0.0210。脑梗死病人群体的C、T等位基因的频率分别为0.8491、0.1509;CC、CT、TT基因型的频率分别为0.7170、0.2641、0.0189。 HIF-1α基因rs2057482 SNP位点多态性在10个两代三口家系调查中均未发现遗传变异。讨论本研究采用双向等位基因特异性扩增技术,对HIF-1α基因rs2057482位点多态性在辽宁汉族、广西壮族群体多态性分布情况进行分析,结果表明rs2057482位点多态性在辽宁汉族与广西壮族群体的基因频率分布具有显著性差异(P＜0.05),与韩国人群的基因频率调查数据进行比较,结果显示韩国人群与广西壮族群体之间具有显著性差异(P＜0.05)。而与辽宁汉族群体的等位基因频率较为接近,差异无统计学意义(P＞0.05)。提示HIF-1α基因rs2057482位点多态性在不同群体中的分布存在差异性。关于rs2057482位点在其他群体的分布情况目前未见报道。近年来,随着HIF-1α在中枢神经系统中的作用不断被认识,Marti等通过对小鼠脑缺血动物模型的研究表明:脑缺血缺氧可导致HIF-1的表达增加,激活血管内皮生长因子及它的受体基因,两者相结合引起脑组织的血管内皮细胞增殖,并且诱导新生血管形成,改善血、氧供给不足造成的神经损害,同时也抑制了缺氧诱导的神经细胞的凋亡,这一生理机理可能改善脑缺血患者的预后。表明HIF-1α与脑缺血的关系密切,在脑缺血的保护性反应中起重要作用。脑梗死是脑缺血缺氧性疾病常见的病理变化。本研究结果显示HIF-1αrs2057482位点在健康对照组与脑梗死病例组之间的基因及基因型频率分布无显著性差异(P＞0.05),提示HIF-1αrs2057482位点多态性与脑梗死无相关性。本文根据对辽宁汉族、广西壮族两个群体的基因频率调查数据计算法医学参数,HIF-1α基因rs2057482位点在辽宁汉族人群和广西壮族人群中的杂合度(H)分别为0.564和0.350;多态性信息含量(PIC)分别为0.2423和0.1597;个人识别率(DP)分别为0.445和0.292;非父排除率(EPP)分别为0.121和0.080。该位点在辽宁汉族群体中rs2057482位点属于中等鉴别能力类遗传标记,具有法医学应用价值。结论 1、HIF-1α基因rs2057482 SNP位点在辽宁汉族、广西壮族及脑梗死病人群体中均具有遗传多态性,基因频率分布符合Hardy-Weinberg平衡。 2、广西壮族群体的HIF-1α基因rs2057482位点基因分布与辽宁汉族和韩国人群具有显著性差异;辽宁汉族群体与韩国人群之间频率分布无统计学差异。 3、HIF-1α基因rs2057482位点与脑梗死无相关性。 4、HIF-1α基因rs2057482 SNP位点在辽宁汉族属于中等鉴别能力的遗传标记,在法医学亲子鉴定和个人识别中具有应用价值。
[Abstract]:Foreword

Hypoxia - inducible factor - 1 ( HIF - 1 ) is the only transcription factor which plays an active role in hypoxia and hypoxia .

It is found that the single nucleotide polymorphism of HIF - 1伪 is closely related to diabetes , myocardial ischemia infarction and non - traumatic femoral head necrosis .

The rs2057482 SNP site of HIF - 1伪 gene was selected to investigate the single nucleotide polymorphism of rs2057482 locus in Han nationality and Guangxi Zhuang nationality in Liaoning province . The relationship between the gene and cerebral infarction disease was discussed . It was designed to provide valuable reference data for human genetics , clinical medicine and forensic medicine .

Materials and Methods

The blood samples of 1,259 Han and 191 healthy blood donors and 159 patients with cerebral infarction were collected from the Han population of Liaoning , and 30 DNA samples were collected from 10 2 generation 3 families .

2 . The distribution of rs2057482SNP loci of HIF - 1伪 and rs2057482SNP was investigated by bi - directional allele specific amplification ( Bi - ASA ) .

3 . The genotype and allele frequencies were calculated by direct counting method . The results showed that the population was representative ; the statistical software of SPSS 15.0 was used to analyze the gene distribution of different population and the analysis between the control group and the case group . The parameters of forensic medicine were calculated as heterozygosity ( H ) , polymorphism information content ( PIC ) , personal identification rate ( DP ) and non - parent exclusion rate ( epp ) .

Results

Among the three groups , the C and T alleles of the rs2057482 locus of HIF - 1伪 gene were detected , and three genotypes of CC , CT and TT were detected .

The frequencies of C and T allele were 0.6873 , 0.2857 and 0.0270 respectively in the Han population of Liaoning . The frequencies of C and T allele were 0.9031 , 0.0969 , 0.0210 respectively . The frequencies of C and T allele were 0.8491 and 0.915 respectively . The frequencies of CC , CT and TT genotype were 0.7170 , 0.2641 and 0.0189 , respectively .

The SNP site polymorphism of HIF - 1伪 gene rs2057482 did not find any genetic variation in the investigation of 10 generations of three families .

discuss

The polymorphism of rs2057482 polymorphism of HIF - 1伪 gene was analyzed in the Han nationality and Guangxi Zhuang ethnic group . The results showed that the polymorphism of rs2057482 polymorphism was significantly different between the Han population and Zhuang population in Liaoning ( P < 0.05 ) . The results showed that the polymorphism of rs2057482 polymorphism in the Han population was different from that of the Han population in Liaoning ( P > 0.05 ) . The distribution of rs2057482 locus in other groups was not reported .

In recent years , with the increasing role of HIF - 1伪 in the central nervous system , Marti et al . have shown that the expression of HIF - 1 can be increased , vascular endothelial growth factor ( VEGF ) and its receptor gene can be activated by cerebral ischemia and hypoxia .

The heterozygosity ( H ) of HIF - 1伪 gene rs2057482 was 0.564 and 0.350 respectively in the Han population and Guangxi Zhuang nationality in Liaoning Han population . The polymorphism information content ( PIC ) was 0.2423 and 0.1597 , 0.121 and 0.080 , respectively .

Conclusion

1 . The SNP locus of HIF - 1伪 gene rs2057482 has a genetic polymorphism in the population of Han , Guangxi Zhuang and cerebral infarction patients in Liaoning .

2 . The distribution of HIF - 1伪 gene rs2057482 in Guangxi Zhuang ethnic group was significantly different from that of Han nationality and Korean population in Liaoning province . There was no statistical difference between Han population and Korean population in Liaoning .

3 . The rs2057482 locus of HIF - 1伪 gene had no correlation with cerebral infarction .

4 . The SNP locus of HIF - 1伪 gene rs2057482 is a genetic marker of moderate discrimination in the Han nationality in Liaoning , and has application value in forensic paternity testing and personal identification .
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：D919

【参考文献】