PRDM16基因及母亲孕期环境暴露因素与非综合征型唇腭裂相关性研究

doi:10.7518/hxkq.2018.05.008

摘要/Abstract

摘要：

目的探究PRDM16基因rs7525173、rs2236518、rs2493264及母亲孕早期吸烟饮酒与非综合征型唇腭裂（NSCL/P）发生的相关性。方法收集157个患者-父母核心家系,采用连接酶检测反应（LDR）和直接测序两种方法进行基因分型,使用传递不平衡检验（TDT）、连锁不平衡检验（LD）等对数据进行统计分析。收集1 710例唇腭裂患者及956例健康新生儿,填写《唇腭裂患者流行病学调查问卷》,对孕期父母吸烟饮酒暴露因素进行分析。结果 rs2236518位点C等位基因在腭裂组中存在过传递（P<0.05）,其余位点在各组中均无明显统计学意义。母亲吸烟、母亲被动吸烟及母亲饮酒3个因素存在统计学差异（P<0.05）。结论 PRDM16基因rs2236518多态性与NSCL/P存在相关性,母亲吸烟、母亲被动吸烟及母亲饮酒与唇腭裂的发生存在密切联系。

关键词: 非综合征型唇腭裂, PRDM16基因, 单核苷酸多态性, 传递不平衡检验, 连锁不平衡检验

Abstract:

Objective We aimed to study the association between rs7525173, rs2236518, rs2493264 single nucleotide poly-morphism (SNP) in the PRDM16 gene, smoking, alcohol exposures, and nonsyndromic cleft lip with or without cleft palate (NSCL/P). Methods A total of 157 case-parent trios were selected, and SNPs were genotyped by using ligase detection reaction (LDR) and direct sequencing methods. Transmission disequilibrium test (TDT) and linkage disequilibrium (LD) tests were con-ducted to analyze the data. A total of 1 710 patients with orofacial clefts and 956 healthy newborns were enrolled in the epide-miological survey. The smoking and drinking exposures of parents during early pregnancy were analyzed. Results The C allele at rs2236518 was over-transmitted for NSCPO (P<0.05). Statistical differences were observed among three factors, namely, maternal smoking, maternal passive smoking, and maternal drinking (P<0.05). Conclusion The rs2236518 at PRDM16 gene, maternal smoking, maternal passive smoking, and maternal drinking were closely related to the occurrence of NSCL/P.

Key words: nonsyndromic cleft lip with or without cleft palate, PRDM16 gene, single nucleotide polymorphism, trans-mission disequilibrium test, linkage disequilibrium

中图分类号:

R782.21

殷斌,石冰,贾仲林. PRDM16基因及母亲孕期环境暴露因素与非综合征型唇腭裂相关性研究[J]. 华西口腔医学杂志, 2018, 36(5): 503-507.

Bin Yin,Bing Shi,Zhonglin Jia. Associations among PRDM16 polymorphisms, environmental exposure factors during mother’s pregnancy, and non-syndromic cleft lip with or without cleft palate[J]. West China Journal of Stomatology, 2018, 36(5): 503-507.

图/表 7

表 1

表 2

表 3

表 4

表 5

表 6

表 7

参考文献 23

[1]	Shkoukani MA, Chen M, Vong A . Cleft lip—a comprehen-sive review[J]. Front Pediatr, 2013,1:53.
[2]	Forrester MB, Merz RD . Descriptive epidemiology of oral clefts in a multiethnic population, Hawaii, 1986—2000[J]. Cleft Palate Craniofac J, 2004,41(6):622-628. doi: 10.1597/03-089.1 URL pmid: 15516165
[3]	王翰章 . 发展我国特色的唇腭裂治疗方法[J]. 口腔颌面外科杂志, 2008,18(1):1-5. doi: 10.3969/j.issn.1005-4979.2008.01.001 URL
	Wang HZ . Development in Chinese way of treatment to cleft lip and palate[J]. J Oral Maxillofac Surg, 2008,18(1):1-5. doi: 10.3969/j.issn.1005-4979.2008.01.001 URL
[4]	Boehringer S, van der Lijn F, Liu F , et al. Genetic determi-nation of human facial morphology: links between cleft-lips and normal variation[J]. Eur J Hum Genet, 2011,19(11):1192-1197. doi: 10.1038/ejhg.2011.110 URL pmid: 21694738
[5]	Edwards M, Roddick L, Scott R . Autosomal dominantnon-syndromic cleft lip and palate linked to chromosome 4[J]. Am J Hum Genet Suppl, 2003,73:672.
[6]	Levinson DF, Levinson MD, Segurado R , et al. Genome scan meta-analysis of schizophrenia and bipolar disorder, partⅠ: Methods and power analysis[J]. Am J Hum Genet, 2003,73(1):17-33. doi: 10.1086/376549 URL pmid: 12802786
[7]	Field LL, Ray AK, Cooper ME , et al. Genome scan for loci involved in nonsyndromic cleft lip with or without cleft palate in families from West Bengal, India[J]. Am J Med Genet A, 2004,130A(3):265-271. doi: 10.1002/(ISSN)1096-8628 URL
[8]	Marazita ML, Field LL, Cooper ME , et al. Genome scan for loci involved in cleft lip with or without cleft palate, in Chinese multiplex families[J]. Am J Hum Genet, 2002,71(2):349-364. doi: 10.1086/341944 URL
[9]	Marazita ML, Field LL, Tunçbilek G , et al. Genome-scan for loci involved in cleft lip with or without cleft palate in consanguineous families from Turkey[J]. Am J Med Genet A, 2004,126A(2):111-122. doi: 10.1002/ajmg.a.v126a:2 URL
[10]	Neiswanger K, Ford MD, Cooper ME , et al. Genome scan of cleft lip with or without cleft palate (CL/P).Ⅱ. Broadening the phenotype to include velopharyngeal incompetence (VPI)[J]. Am J Hum Genet, 2003,73(Suppl):493.
[11]	Moreno LM, Arcos-Burgos M, Marazita ML , et al. Genetic analysis of candidate loci in non-syndromic cleft lip families from Antioquia-Colombia and Ohio[J]. Am J Med Genet A, 2004,125A(2):135-144. doi: 10.1002/(ISSN)1096-8628 URL
[12]	Bjork BC, Turbe-Doan A, Prysak M , et al. PRDM16 is required for normal palatogenesis in mice[J]. Hum Mol Genet, 2010,19(5):774-789. doi: 10.1093/hmg/ddp543 URL pmid: 2816611
[13]	Warner DR, Mukhopadhyay P, Webb CL , et al. Chromatin immunoprecipitation-promoter microarray identification of genes regulated by PRDM16 in murine embryonic palate mesenchymal cells[J]. Exp Biol Med(Maywood), 2012,237(4):387-394. doi: 10.1258/ebm.2012.011258 URL
[14]	Hackshaw A, Rodeck C, Boniface S . Maternal smoking in pregnancy and birth defects: a systematic review based on 173 687 malformed cases and 11.7 million controls[J]. Hum Reprod Update, 2011,17(5):589-604. doi: 10.1093/humupd/dmr022 URL
[15]	Marshall EG, Harris G, Wartenberg D . Oral cleft defects and maternal exposure to ambient air pollutants in New Jersey[J]. Birth Defects Res A Clin Mol Teratol, 2010,88(4):205-215. doi: 10.1002/bdra.20650 URL pmid: 2862481
[16]	Wyszynski DF, Wu T . Use of US birth certificate data to estimate the risk of maternal cigarette smoking for oral clefting[J]. Cleft Palate Craniofac J, 2002,39(2):188-192. doi: 10.1597/1545-1569(2002)039<0188:UOUSBC>2.0.CO;2 URL
[17]	Gorlin RJ, Cohen MM, Hennekam RCM. Syndromes of the head and neck[M]. Oxford: Oxford University Press, 2001.
[18]	Warner DR, Horn KH, Mudd L , et al. A novel Smad binding protein expressed in murine embryonic orofacial tissue[J]. Biochim Biophys Acta, 2007,1773(6):814-820. doi: 10.1016/j.bbamcr.2007.03.016 URL pmid: 17467076
[19]	Warner DR, Wells JP, Greene RM , et al. Gene expression changes in the secondary palate and mandible of PRDM16(-/-) mice[J]. Cell Tissue Res, 2013,351(3):445-452. doi: 10.1007/s00441-012-1525-2 URL pmid: 23149718
[20]	Bjork BC, Vieira AR, Faust S , et al. Phenotypic, genetic, and developmental characterization of CPO1, a recessive ENU-induced mouse model of cleft palate[M]. Woodbury, NY: Mouse Molecular Genetics Cold Spring Harbor Press, 2006: 27.
[21]	Mirilas P, Mentessidou A, Kontis E , et al. Parental expo-sures and risk of nonsyndromic orofacial clefts in offspring: a case-control study in Greece[J]. Int J Pediatr Otorhino-laryngol, 2011,75(5):695-699. doi: 10.1016/j.ijporl.2011.02.018 URL
[22]	Munger RG, Romitti PA, Daack-Hirsch S , et al. Maternal alcohol use and risk of orofacial cleft birth defects[J]. Tera-tology, 1996,54(1):27-33. doi: 10.1002/(SICI)1096-9926(199607)54:13.0.CO;2-0 URL pmid: 8916367
[23]	舒申友, 唐世杰, 吴燊荣 , 等. 广东人群非综合征型唇腭裂发病危险因素研究[J]. 中国修复重建外科杂志, 2010,24(8):962-966.
	Shu SY, Tang SJ, Wu SR , et al. Risk factors for nonsyn-dromic cleft lip and palate in Guangdong population[J]. Chin J Reparat Reconstruct Surg, 2010,24(8):962-966.

SNP	等位基因	PCR引物序列（5’—3’）	扩增长度/bp	探针序列
rs7525173	C/G	F：TCTGGCCAAGAAAACACCC	100	TTTTTTTTTTTTTTTTTTTTTTTTAAGTAGTTCAGAA-
				ACGCATCGGG
		R：TCACCCGCAGGCTCTTATTC		TTTTTTTTTTTTTTTTTTTTTTTTTTAAGTAGTTCAGA-
				AACGCATCGGC
rs2236518	A/C	F：CTTTTCAGCAGGAGCAGAAC	82	TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTGGGGGATC-
				CGGAGACCAACGGT
		R：AAAACGGCGGGATGGGACTG		TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTGGGGGA-
				TCCGGAGACCAACGGG
rs2493264	C/T	F：GGCAGGGGACCACAATCCA	99	TTTTTTTTTTTTTTTTTTTTCGTATTTTAGACTCGAAG-
				CCTCG
		R：GTCTGCAAAACACTTGCCTG		TTTTTTTTTTTTTTTTTTTTTTCGTATTT

SNPs	D’值	95%可信区间	χ²值
rs7525173与rs2236518	0.035	-0.01,0.23	0.000
rs7525173与rs2493264	0.185	0.01,0.52	0.001
rs2236518与rs2493264	0.695	0.57,0.79	0.140

类型	SNP	等位基因	MAF	T/U	χ²值	P值
NSCL/P	rs7525173	G	0.10	28︰21	1.0	0.317 3
	rs2236518	C	0.42	56︰51	0.234	0.628 8
	rs2493264	T	0.18	36︰34	0.057	0.811 0
NSCLP	rs7525173	G	0.10	35︰28	0.778	0.377 8
	rs2236518	C	0.42	76︰67	0.566	0.451 7
	rs2493264	T	0.18	51︰47	0.163	0.686 2
NSCPO	rs7525173	G	0.10	7︰7	0.0	1.0
	rs2236518	C	0.42	25︰11	5.444	0.019 6
	rs2493264	T	0.18	15︰13	0.143	0.705 5

因素	病例组		对照组		χ²值	P值
因素	是	否	是	否	χ²值	P值
母亲吸烟	44	1 666	2	954	20.207	0.000
母亲被动吸烟	424	1 286	83	873	103.377	0.000
母亲饮酒	425	1 285	74	882	118.037	0.000
父亲吸烟	859	851	496	460	0.667	0.414
父亲饮酒	1 085	625	575	381	2.849	0.091

危险因素	P值	比值比	95%可信区间
母亲吸烟	0.000	12.598	3.047,52.081
母亲被动吸烟	0.000	3.468	2.700,4.455
母亲饮酒	0.000	3.942	3.035,5.119
父亲吸烟	0.414	0.936	0.799,1.097
父亲饮酒	0.092	1.150	0.978,1.353