染色体9p21单核苷酸多态性与脑梗死的相关性分析

doi:10.3969/j.issn.1673-8640.2015.04.018

摘要/Abstract

摘要：

目的探讨染色体9p21的单核苷酸多态性(SNP)与脑梗死易感性的关系。方法应用聚合酶链反应-高温连接酶检测反应(PCR-LDR)检测528例非心源性脑梗死患者(375例合并颈动脉斑块,153例无颈动脉斑块)、258名正常对照者的染色体9p21.3区带rs10757278、rs1333049、rs2383206、rs1537378、rs4977574和rs2383207 6个位点的SNP。同时采用多元Logistic回归分析和χ²检验对统计有意义的位点分析基因交互作用,计算比值比(OR)和95%可信区间(CI)。结果在45~65岁的中年人群中,rs2383206和rs4977574的G等位基因在脑梗死无颈动脉斑块组的携带人数(分别为55.0%和54.2%)明显多于正常对照组(45.9%和44.4%,P=0.024、0.015);而同时含有rs2383206和rs4977574 2个位点GG纯合突变基因型的个体发生脑梗死的风险可能是同时携带非GG纯合突变基因型(GA+AA)个体的1.733倍。6个位点之间都存在强的连锁不平衡关系。结论染色体9p21区带中rs2383206、rs4977574 2个位点的SNP与急性脑梗死的发生易感性可能有一定关联。

关键词: 单核苷酸多态性, 染色体9p21, 脑梗死

Abstract:

Objective To study the relationship between single nucleotide polymorphism (SNP) in chromosome 9p21 and the susceptibility of cerebral infarction. Methods Polymerase chain reaction-ligase detection reaction (PCR-LDR) was applied to determine the six gene locus of SNP between the observation group and normal control group. These six gene locus were rs10757278, rs1333049, rs2383206, rs1537378, rs4977574 and rs2383207 in chromosome 9p21.3. And the observation group had 528 cases of patients with non cardiogenic cerebral infarction (375 cases of them had carotid artery plaque, and the other 153 cases did not have), while the normal control group had 258 cases. Using multivariate Logistic regression and χ² test to analysis the site of the statistical significant genetic interactions, calculated the odds ratios (OR) and 95% confidence interval (CI). Results At the age of 45-65 middle-aged crowd, G allelic genes of rs2383206 and rs4977574, the carriers without carotid artery plaque in cerebral infarction group (55.0% and 54.2%) were significantly more than the normal control group (45.9% and 44.4%, P=0.024, 0.015);And to the risk of cerebral infarction, the GG genotype homozygous mutations accompanied with two gene locus rs2383206 and rs4977574 was 1.733 times to non-GG genotype homozygous mutations(GA+AA). The six gene locus had strong disequilibrium between relations. Conclusions 9p21 chromosome zone in rs2383206 and rs4977574 two loci gene SNP relate to the susceptibility to the occurrence of acute cerebral infarction.

Key words: Single nucleotide polymorphism, Chromosome 9p21, Cerebral infarction

中图分类号:

Q503

朱扬, 卢钟娇, 傅毅. 染色体9p21单核苷酸多态性与脑梗死的相关性分析[J]. 检验医学, 2015, 30(4): 367-375.

ZHU Yang, LU Zhongjiao, FU Yi. Correlation analysis of cerebral infarction with single nucleotide polymorphism of chromosome 9p21[J]. Laboratory Medicine, 2015, 30(4): 367-375.

图/表 11

参考文献 23

[1]	DELLA-Morte D, GUADAGNI F, PALMIROTTA R, et al.Genetics of ischemic stroke, stroke-related risk factors, stroke precursors and treatments[J]. Pharmacogenomics,2012,13(5):595-613.
[2]	FLOSSMANN E, SCHULZ UG, ROTHWELL PM.Systematic review of methods and results of studies of the genetic epidemiology of ischemic stroke[J]. Stroke,2004,35(1):212-227.
[3]	MCPHERSON R, PERTSEMLIDIS A, KAVASLAR N, et al.A common allele on chromosome 9 associated with coronary heart disease[J]. Science,2007,316(5830):1488-1491.
[4]	SAMANI NJ, ERDMANN J, HALL AS, et al.Genomewide association analysis of coronary artery disease[J]. N Engl J Med,2007,357(5):443-453.
[5]	SCHEFFOLD T, KULLMANN S, HUGE A, et al.Six sequence variants on chromosome 9p21.3 are associated with a positive family history of myocardial infarction: a multicenter registry[J]. BMC Cardiovasc Disord,2011,11(7):9.
[6]	ANDERSON CD, BIFFI A, ROST NS, et al.Chromosome 9p21 in ischemic stroke: population structure and meta-analysis[J]. Stroke,2010,41(6):1123-1131.
[7]	GSCHWENDTNER A, BEVAN S, COLE JW, et al.Sequence variants on chromosome 9p21.3 confer risk for atherosclerotic stroke[J]. Ann Neurol,2009,65(5):531-539.
[8]	ZHANG Z, WANG BJ, GUAN HY, et al.A LDR-PCR approach for multiplex polymorphisms genotyping of severely degraded DNA with fragment sizes<100 bp[J]. J Forensic Sci,2009,54(6):1304-1309.
[9]	HECKMAN MG, SOTO-ORTOLAZA AI, DIEHL NN, et al.Genetic variants associated with myocardial infarction in the PSMA6 gene and Chr9p21 are also associated with ischaemic stroke[J]. Eur J Neurol,2013,20(2):300-308.
[10]	LÖVKVIST H, SJÖGREN M, HÖGLUND P, et al. Are 25 SNPs from the CARDIoGRAM study associated with ischaemic stroke[J]. Eur J Neurol,2013,20(9):1284-1291.
[11]	TRAYLOR M, FARRALL M, HOLLIDAY EG, et al.Genetic risk factors for ischaemic stroke and its subtypes (the METASTROKE Collaboration): a meta-analysis of genome-wide association studies[J]. Lancet Neurol,2012,11(11):951-962.
[12]	SMITH JG, MELANDER O, LÖVKVIST H, et al. Common genetic variants on chromosome 9p21 confers risk of ischemic stroke: a large-scale genetic association study[J]. Circ Cardiovasc Genet,2009,2(2):159-164.
[13]	The group of guidelines for acute ischemic stroke in Chinese Medical Association. Guidelines for acute ischemic stroke in China of 2010[J]. China Medical News,2010,25(14):16-19.
[14]	ADAMS HP, BENDIXEN BH, KAPPELLE LJ, et al.Classification of subtype of acute ischemic stroke. Definitions for use in a multicenter clinical trial. TOAST. Trial of Org 10172 in acute stroke treatment[J]. Stroke, 1993,24(1):35-41.
[15]	STEIN JH, KORCARZ CE, HURST RT, et al.Use of carotid ultrasound to identify subclinical vascular disease and evaluate cardiovascular disease risk: a consensus statement from the American Society of Echocardiography Carotid Intima-Media Thickness Task Force. Endorsed by the Society for Vascular Medicine[J]. J Am Soc Echocardiogr,2008,21(2):93-111.
[16]	SCHULZ UG, FLOSSMANN E, ROTHWELL PM.Heritability of ischemic stroke in relation to age, vascular risk factors, and subtypes of incident stroke in population-based studies[J]. Stroke,2004,35(4):819-824.
[17]	OLSSON S, JOOD K, BLOMSTRAND C, et al.Genetic variation on chromosome 9p21 shows association with the ischaemic stroke subtype large-vessel disease in a Swedish sample aged≤70[J]. Eur J Neurol,2011,18(2):365-367.
[18]	SHI YY, HE L.SHESIS, a powerful software platform for analyses of linkage disequilibrium, haplotype construction, and genetic association at polymorphism loci[J]. Cell Res, 2005,15(2):97-98.
[19]	KRUGLYAK L.Prospects for whole-genome linkage disequilibrium mapping of common disease genes[J]. Nat genet, 1999,22(2):139-144.
[20]	JARINOVA O, STEWART AF, ROBERTS R, et al.Functional analysis of the chromosome 9p21. 3 coronary artery disease risk locus[J]. Arterioscler Tromb Vasc Bio,2009,29(10):1671-1677.
[21]	BURD CE, JECK WR, LIU Y, et al.Expression of linear and novel circular forms of an INK4/ARF-associated non-coding RNA correlates with atherosclerosis risk[J]. PLoS Genet,2010,6(12):e1001233.
[22]	VISEL A, ZHU Y, MAY D, et al.Targeted deletion of the 9p21 non-coding coronary artery disease risk interval in mice[J]. Nature,2010,464(7287):409-412.
[23]	HU WL, LI SJ, LIU DT, et al.Genetic variants on chromosome 9p21 and ischemic stroke in Chinese[J]. Brain Res Bull,2009,79(6):431-435.

组别	年龄 (岁)	男性 [例数(%)]	高脂血症 [例数(%)]	TC (mmol/L)	HDL-C (mmol/L)	LDL-C (mmol/L)
脑梗死组
无颈动脉斑块组	56.56±7.65	96(62.70)	52(34.00)	4.60±1.12	1.22±0.40	2.86±0.90
有颈动脉斑块组	56.34±7.85^*	127(49.20)	79(30.60)	4.68±0.96	1.35±1.42^*	2.80±0.80
正常对照组	60.90±6.24	256(68.30)^*	148(39.50)	4.50±1.33	1.12±0.35	3.02±0.93
组别	TG (mmol/L)	糖尿病 [例数(%)]	空腹血糖 (mmol/L)	高血压 [例数(%)]	收缩压 (kPa)	舒张压 (kPa)
脑梗死组
无颈动脉斑块组	1.90±1.21	23(15.00)	5.78±2.19	81(52.90)	18.4±2.8	11.1±1.6
有颈动脉斑块组	1.91±1.46	29(11.20)^*	5.51±1.46^*	84(36.40)^*	17.6±2.1^*	10.5±1.3
正常对照组	2.04±1.34	86(22.90)	6.24±2.82	246(65.60)^*	18.9±2.5^*	11.1±1.5^*

组别	年龄 (岁)	男性 [例数(%)]	高脂血症 [例数(%)]	TC (mmol/L)	HDL-C (mmol/L)	LDL-C (mmol/L)
脑梗死组
无颈动脉斑块组	56.56±7.65	96(62.70)	52(34.00)	4.60±1.12	1.22±0.40	2.86±0.90
有颈动脉斑块组	56.34±7.85^*	127(49.20)	79(30.60)	4.68±0.96	1.35±1.42^*	2.80±0.80
正常对照组	60.90±6.24	256(68.30)^*	148(39.50)	4.50±1.33	1.12±0.35	3.02±0.93
组别	TG (mmol/L)	糖尿病 [例数(%)]	空腹血糖 (mmol/L)	高血压 [例数(%)]	收缩压 (kPa)	舒张压 (kPa)
脑梗死组
无颈动脉斑块组	1.90±1.21	23(15.00)	5.78±2.19	81(52.90)	18.4±2.8	11.1±1.6
有颈动脉斑块组	1.91±1.46	29(11.20)^*	5.51±1.46^*	84(36.40)^*	17.6±2.1^*	10.5±1.3
正常对照组	2.04±1.34	86(22.90)	6.24±2.82	246(65.60)^*	18.9±2.5^*	11.1±1.5^*

年龄 (岁)	组别	rs10757278		rs1333049		rs2383206
年龄 (岁)	组别	G	A	C	G	G	A
<45	脑梗死组	13(54.2)	11(45.8)	13(54.2)	11(45.8)	13(54.2)	11(45.8)
	正常对照组	14(36.8)	24(63.2)	14(36.8)	24(63.2)	14(36.8)	24(63.2)
	P值	0.376		0.376		0.376
	OR值(95%CI)	1.659(0.541~5.085)		1.658(0.541~5.076)		0.603(0.197~1.848)
45~65	脑梗死组	129(54.2)	109(45.8)	129(54.2)	109(45.8)	131(55.0)	107(45.0)
	正常对照组	191(46.6)	219(53.4)	191(46.6)	219(53.4)	188(45.9)	222(54.1)
	P值	0.052		0.054		0.021
	OR值(95%CI)	1.383(0.997~1.923)		1.381(0.995~1.917)		1.472(1.060~2.044)
>65	脑梗死组	18(40.9)	26(59.1)	18(40.9)	26(59.1)	15(34.1)	29(65.9)
	正常对照组	35(51.5)	33(48.5)	35(51.5)	33(48.5)	35(51.5)	33(48.5)
	P值	0.242		0.276		0.055
	OR值(95%CI)	0.596(0.251~1.419)		0.619(0.262~1.466)		0.418(0.172~1.020)
年龄 (岁)	组别	rs1537378		rs4977574		rs2383207
年龄 (岁)	组别	T	C	G	A	G	A
<45	脑梗死组	4(16.7)	20(83.3)	13(54.2)	11(45.8)	18(75.0)	6(25.0)
	正常对照组	10(26.3)	28(73.7)	14(36.8)	24(63.2)	22(57.9)	16(42.1)
	P值	0.561		0.376		0.170
	OR值(95%CI)	0.664(0.167~2.639)		0.603(0.197~1.848)		2.320(0.698~7.708)
45~65	脑梗死组	30(12.6)	208(87.4)	129(54.2)	109(45.8)	165(69.3)	73(30.7)
	正常对照组	67(16.3)	343(83.7)	182(44.4)	228(55.6)	267(65.1)	143(34.9)
	P值	0.176		0.013		0.239
	OR值(95%CI)	0.722(0.449~1.157)		1.519(1.094~2.110)		1.235(0.869~1.754)
>65	脑梗死组	5(11.4)	39(88.6)	16(36.4)	28(63.6)	26(59.1)	18(40.9)
	正常对照组	8(11.8)	60(88.2)	34(50.0)	34(50.0)	48(70.6)	20(29.4)
	P值	0.712		0.160		0.326
	OR值(95%CI)	0.783(0.214~2.867)		0.534(0.222~1.282)		0.639(0.261~1.562)

年龄 (岁)	组别	rs10757278		rs1333049		rs2383206
年龄 (岁)	组别	G	A	C	G	G	A
<45	脑梗死组	13(54.2)	11(45.8)	13(54.2)	11(45.8)	13(54.2)	11(45.8)
	正常对照组	14(36.8)	24(63.2)	14(36.8)	24(63.2)	14(36.8)	24(63.2)
	P值	0.376		0.376		0.376
	OR值(95%CI)	1.659(0.541~5.085)		1.658(0.541~5.076)		0.603(0.197~1.848)
45~65	脑梗死组	129(54.2)	109(45.8)	129(54.2)	109(45.8)	131(55.0)	107(45.0)
	正常对照组	191(46.6)	219(53.4)	191(46.6)	219(53.4)	188(45.9)	222(54.1)
	P值	0.052		0.054		0.021
	OR值(95%CI)	1.383(0.997~1.923)		1.381(0.995~1.917)		1.472(1.060~2.044)
>65	脑梗死组	18(40.9)	26(59.1)	18(40.9)	26(59.1)	15(34.1)	29(65.9)
	正常对照组	35(51.5)	33(48.5)	35(51.5)	33(48.5)	35(51.5)	33(48.5)
	P值	0.242		0.276		0.055
	OR值(95%CI)	0.596(0.251~1.419)		0.619(0.262~1.466)		0.418(0.172~1.020)
年龄 (岁)	组别	rs1537378		rs4977574		rs2383207
年龄 (岁)	组别	T	C	G	A	G	A
<45	脑梗死组	4(16.7)	20(83.3)	13(54.2)	11(45.8)	18(75.0)	6(25.0)
	正常对照组	10(26.3)	28(73.7)	14(36.8)	24(63.2)	22(57.9)	16(42.1)
	P值	0.561		0.376		0.170
	OR值(95%CI)	0.664(0.167~2.639)		0.603(0.197~1.848)		2.320(0.698~7.708)
45~65	脑梗死组	30(12.6)	208(87.4)	129(54.2)	109(45.8)	165(69.3)	73(30.7)
	正常对照组	67(16.3)	343(83.7)	182(44.4)	228(55.6)	267(65.1)	143(34.9)
	P值	0.176		0.013		0.239
	OR值(95%CI)	0.722(0.449~1.157)		1.519(1.094~2.110)		1.235(0.869~1.754)
>65	脑梗死组	5(11.4)	39(88.6)	16(36.4)	28(63.6)	26(59.1)	18(40.9)
	正常对照组	8(11.8)	60(88.2)	34(50.0)	34(50.0)	48(70.6)	20(29.4)
	P值	0.712		0.160		0.326
	OR值(95%CI)	0.783(0.214~2.867)		0.534(0.222~1.282)		0.639(0.261~1.562)

组别	GG^*±GG^#	(GA+AA)^*+GG^#	GG^*+(GA+AA)^#	(GA+AA)^*+(GA+AA)^#
无颈动脉斑块的脑梗死组	40(26.1)	1(0.7)	2(1.3)	110(71.9)
正常对照组	46(17.8)	1(0.4)	5(1.9)	206(79.8)
OR值(95%CI)		1.271(0.072~22.220)	3.174(0.566~17.860)	1.733(1.056~2.849)
P值		0.870	0.189	0.030