本文選題：HIF-1α基因 + 單核苷酸多態(tài)性　； 參考：《中國醫(yī)科大學(xué)》2009年碩士論文

【摘要】： 前言 缺氧誘導(dǎo)因子1(hypoxia-inducible factor-1,HIF-1)是目前發(fā)現(xiàn)的唯一特異性缺氧狀態(tài)下發(fā)揮活性的轉(zhuǎn)錄因子,于1992年首先由Semenza等在低氧的肝細(xì)胞癌細(xì)胞株Hep3B細(xì)胞的核提取物中發(fā)現(xiàn)。HIF-1廣泛存在于哺乳動(dòng)物和人體內(nèi),是由α和β兩個(gè)亞單位組成的一種異源二聚體。其中HIF-1α是唯一的氧調(diào)節(jié)亞單位,只有在低氧或缺氧情況下才表現(xiàn)其生物學(xué)活性,它主要通過與靶基因的缺氧反應(yīng)元件(hypoxia response element,HRE)上的HIF-1結(jié)合位點(diǎn)(5'-TACGTG CT-3')相結(jié)合,從而激活靶基因的轉(zhuǎn)錄,發(fā)揮一系列的生物學(xué)功能,其中包括參與血管生成、能量代謝、紅細(xì)胞生成、細(xì)胞增殖和存活、血管重塑以及血管舒縮反應(yīng)等。 編碼HIF-1α的基因位于14號(hào)染色體q21-24區(qū),該區(qū)域單核苷酸多態(tài)性十分豐富,截止目前,NCBI已公布了238個(gè)SNPs位點(diǎn),其中研究較多的是編碼區(qū)的SNPs位點(diǎn)。研究發(fā)現(xiàn)此區(qū)域的單核苷酸多態(tài)性可以影響蛋白質(zhì)表達(dá)和功能,國外研究多集中在HIF1α基因遺傳多態(tài)性與各種腫瘤(如泌尿、消化、生殖等系統(tǒng)腫瘤)關(guān)系研究,也有報(bào)道與糖尿病、心肌缺血梗死以及非創(chuàng)傷性股骨頭壞死等疾病密切相關(guān)。而HIF-1α基因在中國遼寧漢族、廣西壯族人群的分布及其法醫(yī)學(xué)應(yīng)用價(jià)值國內(nèi)外尚無報(bào)道。另外,研究發(fā)現(xiàn)HIF-1α與腦缺血的關(guān)系密切,在腦缺血的保護(hù)性反應(yīng)中起重要作用,目前國內(nèi)外尚無HIF-1α的基因多態(tài)性與腦缺血缺氧性疾病相關(guān)性的研究報(bào)道。 本研究選擇HIF-1α基因rs2057482 SNP位點(diǎn),對遼寧漢族和廣西壯族人群rs2057482位點(diǎn)單核苷酸多態(tài)性進(jìn)行基因分布調(diào)查,并對該基因與腦梗死疾病的關(guān)系進(jìn)行了探討,旨在為人類遺傳學(xué)、臨床醫(yī)學(xué)以及法醫(yī)學(xué)等領(lǐng)域提供有價(jià)值的參考數(shù)據(jù)。 材料與方法 1、259份遼寧漢族、191份廣西壯族無血緣關(guān)系健康獻(xiàn)血者及159份腦梗死患者的血液樣本。10個(gè)2代3口家系共30份DNA樣品。 2、應(yīng)用雙向等位基因特異性擴(kuò)增(Bi-ASA)技術(shù)調(diào)查HIF-1α基因rs2057482SNP位點(diǎn)在遼寧漢族、廣西壯族健康人群以及腦梗死患者中的分布狀況。并進(jìn)行家系調(diào)查。 3、直接計(jì)數(shù)法計(jì)算基因型及等位基因頻率;Hardy-Weinberg平衡檢驗(yàn)所研究群體是否具有代表性;應(yīng)用SPSS15.0統(tǒng)計(jì)軟件進(jìn)行不同人群基因分布差異以及對照組與病例組組間分析。計(jì)算法醫(yī)學(xué)參數(shù):雜合度(H)、多態(tài)性信息含量(PIC)、個(gè)人識(shí)別率(DP)和非父排除率(EPP)。 結(jié)果 在三個(gè)群體中,均檢測出HIF-1α基因rs2057482位點(diǎn)的C、T兩個(gè)等位基因,及CC、CT、TT三種基因型。 在遼寧漢族群體中,C、T等位基因頻率分別為0.8301、0.1699;CC、CT、TT基因型頻率分別為0.6873、0.2857、0.0270。廣西壯族群體中C、T基因頻率分別為0.9031,0.0969;CC、CT、TT的頻率分別為0.8272、0.1518、0.0210。腦梗死病人群體的C、T等位基因的頻率分別為0.8491、0.1509;CC、CT、TT基因型的頻率分別為0.7170、0.2641、0.0189。 HIF-1α基因rs2057482 SNP位點(diǎn)多態(tài)性在10個(gè)兩代三口家系調(diào)查中均未發(fā)現(xiàn)遺傳變異。 討論 本研究采用雙向等位基因特異性擴(kuò)增技術(shù),對HIF-1α基因rs2057482位點(diǎn)多態(tài)性在遼寧漢族、廣西壯族群體多態(tài)性分布情況進(jìn)行分析,結(jié)果表明rs2057482位點(diǎn)多態(tài)性在遼寧漢族與廣西壯族群體的基因頻率分布具有顯著性差異(P＜0.05),與韓國人群的基因頻率調(diào)查數(shù)據(jù)進(jìn)行比較,結(jié)果顯示韓國人群與廣西壯族群體之間具有顯著性差異(P＜0.05)。而與遼寧漢族群體的等位基因頻率較為接近,差異無統(tǒng)計(jì)學(xué)意義(P＞0.05)。提示HIF-1α基因rs2057482位點(diǎn)多態(tài)性在不同群體中的分布存在差異性。關(guān)于rs2057482位點(diǎn)在其他群體的分布情況目前未見報(bào)道。 近年來,隨著HIF-1α在中樞神經(jīng)系統(tǒng)中的作用不斷被認(rèn)識(shí),Marti等通過對小鼠腦缺血?jiǎng)游锬Ｐ偷难芯勘砻?腦缺血缺氧可導(dǎo)致HIF-1的表達(dá)增加,激活血管內(nèi)皮生長因子及它的受體基因,兩者相結(jié)合引起腦組織的血管內(nèi)皮細(xì)胞增殖,并且誘導(dǎo)新生血管形成,改善血、氧供給不足造成的神經(jīng)損害,同時(shí)也抑制了缺氧誘導(dǎo)的神經(jīng)細(xì)胞的凋亡,這一生理機(jī)理可能改善腦缺血患者的預(yù)后。表明HIF-1α與腦缺血的關(guān)系密切,在腦缺血的保護(hù)性反應(yīng)中起重要作用。腦梗死是腦缺血缺氧性疾病常見的病理變化。本研究結(jié)果顯示HIF-1αrs2057482位點(diǎn)在健康對照組與腦梗死病例組之間的基因及基因型頻率分布無顯著性差異(P＞0.05),提示HIF-1αrs2057482位點(diǎn)多態(tài)性與腦梗死無相關(guān)性。 本文根據(jù)對遼寧漢族、廣西壯族兩個(gè)群體的基因頻率調(diào)查數(shù)據(jù)計(jì)算法醫(yī)學(xué)參數(shù),HIF-1α基因rs2057482位點(diǎn)在遼寧漢族人群和廣西壯族人群中的雜合度(H)分別為0.564和0.350;多態(tài)性信息含量(PIC)分別為0.2423和0.1597;個(gè)人識(shí)別率(DP)分別為0.445和0.292;非父排除率(EPP)分別為0.121和0.080。該位點(diǎn)在遼寧漢族群體中rs2057482位點(diǎn)屬于中等鑒別能力類遺傳標(biāo)記,具有法醫(yī)學(xué)應(yīng)用價(jià)值。 結(jié)論 1、HIF-1α基因rs2057482 SNP位點(diǎn)在遼寧漢族、廣西壯族及腦梗死病人群體中均具有遺傳多態(tài)性,基因頻率分布符合Hardy-Weinberg平衡。 2、廣西壯族群體的HIF-1α基因rs2057482位點(diǎn)基因分布與遼寧漢族和韓國人群具有顯著性差異;遼寧漢族群體與韓國人群之間頻率分布無統(tǒng)計(jì)學(xué)差異。 3、HIF-1α基因rs2057482位點(diǎn)與腦梗死無相關(guān)性。 4、HIF-1α基因rs2057482 SNP位點(diǎn)在遼寧漢族屬于中等鑒別能力的遺傳標(biāo)記,在法醫(yī)學(xué)親子鑒定和個(gè)人識(shí)別中具有應(yīng)用價(jià)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偽 is closely related to diabetes , myocardial ischemia infarction and non - traumatic femoral head necrosis .



The rs2057482 SNP site of HIF - 1偽 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偽 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偽 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偽 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偽 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偽 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偽 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偽 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偽 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偽 gene had no correlation with cerebral infarction .



4 . The SNP locus of HIF - 1偽 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 .
【學(xué)位授予單位】：中國醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：D919

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相關(guān)期刊論文 前1條

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本文編號(hào)：2065904