非综合征性唇腭裂的染色体基因位点研究进展

唇腭裂是常见的先天畸形之一,遗传因素在其发生中发挥着极其重要的作用.随着分子遗传学的发展,越来越多的易感基因被发现.本文就与非综合征性唇腭裂相关的染色体基因位点、与单纯性腭裂相关的染色体基因位点研究进展作一综述.     

宁夏回汉族人群Wnt3基因单核苷酸多态性与非综合征型唇腭裂相关性的研究

目的 探讨宁夏回汉族人群中Wnt3基因rs142167和rs7216231位点单核苷酸多态性（SNP）与非综合征型唇腭裂（NSCL/P）的相关性。方法 收集宁夏地区回汉族人群非综合征型唇腭裂患者371例为病例组,其中汉族患者166例,回族患者205例;收集患者父亲196例,患者母亲224例,其中150例患者为NSCL/P核心家系;258例健康新生儿为对照组,其中汉族190例,回族68例。采用聚合酶链反应-限制性片段长度多态性（PCR-RFLP）方法检测Wnt3基因多态位点rs142167和rs7216231基因型,对比分析2组的基因型和等位基因,并进行传递不平衡检验（TDT）和以家系为基础的相关性检验（FBAT）分析。结果 回汉族人群病例组与对照组比较及其民族分层比较,唇裂、腭裂、唇腭裂及总病例组rs142167和rs7216231位点均无统计学差异（P＞0.05）。TDT分析结果显示：rs142167和rs7216231位点的等位基因均不存在过传递（P＞0.05）。FBAT分析结果显示：单倍型G-G具有统计学意义（P<0.05）。结论 Wnt3基因多态性与宁夏地区回汉族人群非综合征型唇腭裂不存在相关性。     

染色体基因位点与非综合征性唇腭裂致病机制的关系

非综合征性唇腭裂是人类最常见的先天性畸形之一,是一种多基因多因素的遗传疾病.近二十年来,学者们先后在多条染色体上发现了与非综合征性唇腭裂有关的基因位点.本文就染色体基因位点与非综合征性唇腭裂致病机制的关系作一综述.     

Association between MAFB rs17820943 and rs6072081 polymorphism and risk of nonsyndromic cleft lip with or without cleft palate: a meta-analysis

While there have been previous studies examining the relation between the rs17820943 and rs6072081 polymorphisms in the v-maf musculoaponeurotic fibrosarcoma oncogene homolog B (MAFB) gene and rates of nonsyndromic cleft lip with or without cleft palate (NSCL/P), at present the results of these studies have been inconsistent. This meta-analysis therefore aimed to conduct a more robust assessment of the association between the MAFB rs17820943 and rs6072081 polymorphisms and NSCL/P risk. The Embase, Web of Science, PubMed, the China National Knowledge Internet (CNKI), and Wanfang databases were systematically searched to identify relevant studies. In total, five studies incorporating 2769 patients and 2885 controls were identified assessing the rs17820943 polymorphism and three studies incorporating 1242 patients and 1310 controls assessing the rs6072081 polymorphism were identified. This analysis revealed the MAFB rs17820943 and rs6072081 polymorphisms to be linked to a significantly reduced NSCL/P risk (rs17820943: C vs T: OR = 0.76, 95% CI = 0.70-0.82; CC vs CT: OR = 0.75, 95% CI = 0.67-0.85; CC vs TT: OR = 0.58, 95% CI = 0.49-0.67; CC + CT vs TT: OR = 0.67, 95% CI = 0.59-0.77; CT + TT vs CC: OR = 1.43, 95% CI = 1.28-1.60; rs6072081: A vs G: OR = 0.77, 95%CI = 0.68-0.86; AA vs AG: OR = 0.76, 95%CI = 0.64-0.90; AA vs GG: OR = 0.58, 95%CI = 0.45-0.74; AA + AG vs GG: OR = 0.68, 95%CI = 0.54-0.84; AG + GG vs AA: OR = 1.40, 95% CI = 1.19-1.65). The results of the present meta-analysis indicate that in an East Asian population, for both rs17820943 and rs6072081 were associated with NSCL/P.     

Gene-gene interaction for nonsyndromic cleft lip with or without cleft palate in Chilean case-parent trios

ObjectiveNonsyndromic cleft lip with or without cleft palate (NSCL/P) is a birth defect for which several genes susceptibility genes been proposed. Consequently, it has been suggested that many of these genes belong to common inter-related pathways during craniofacial development gene-gene interaction. We evaluated the presence of gene-gene interaction for single nucleotide polymorphisms within interferon regulatory factor 6 (IRF6), muscle segment homeobox 1 (MSX1), bone morphogenetic protein 4 (BMP4) and transforming growth factor 3 (TGFB3) genes in NSCL/P risk in Chilean case-parent trios.DesignFrom previous studies, we retrieved genotypes for 13 polymorphic variants within these four genes in 152 case-parent trios. Using the trio package (R) we evaluate the gene-gen interaction in genetic markers pairs applying a 1°-of-freedom test (1df) and a confirmatory 4°-of-freedom (4df) test for epistasis followed by both a permutation test and a Benjamini-Hochberg test for multiple comparisons adjustment.ResultsWe found evidence of gene-gene interaction for rs6446693 (MSX1) and rs2268625 (TGFB3) (4df p = 0.024; permutation p = 0.015, Benjamini-Hochberg p = 0.001).ConclusionsA significant gene-gene interaction was detected for rs6446693 (MSX1) and rs2268625 (TGFB3). This finding is concordant with research in animal models showing that MSX1 and TGFB3 are expressed in common molecular pathways acting in an epistatic manner during maxillofacial development.     

非综合征性唇腭裂与同源异型盒基因1多态性的相关性研究

目的采用聚合酶链反应- 单链构象多态性（PCR- SSCP）方法研究同源异型盒基因（MSX）1外显子1的编码区，探讨非综合征性唇腭裂（NSCL/P）患者MSX1基因外显子1的编码区内是否存在基因突变。方法采用聚合酶链反应（PCR）和单链构象多态性（SSCP）方法，以45名健康人为对照组，45名NSCL/P患者作为研究对象，分析MSX1基因多态性。结果SSCP分析显示NSCL/P患者（45名）与对照组（45名）样本的电泳速率相同，提示无多态性存在。结论MSX1基因外显子1未发现多态性的存在，其与NSCL/P患者之间无明显相关性。     

中国北方人群非综合征性唇腭裂与BMP4基因多态性的关系

目的探讨BMP4基因rs17563多态性与中国北方人群非综合征性唇腭裂的相关性。方法使用聚合酶链反应-限制性内切酶片段长度多态性分析(PCR-RFLP)方法,在116例非综合征性唇腭裂患者和123名健康对照中,对BMP4基因单核苷酸多态性进行检测。利用SPSS13.0软件分析BMP4基因多态性与非综合征性唇腭裂的相关性。结果 BMP4基因型频率分布符合Hardy-Weinberg平衡。在非综合征性唇腭裂组中CC纯合子明显高于健康对照组,差异有统计学意义(P<0.001)。结论 BMP4基因rs17563多态性可能与中国北方人群非综合征性唇腭裂的发生相关。     

Association between BMP4 gene polymorphisms and cleft lip with or without cleft palate in a population from South China

ObjectivePrevious studies have suggested an association between several polymorphisms of the BMP4 gene and susceptibility to non-syndromic cleft lip with or without cleft palate (NSCL/P) in various populations. However, this association may vary according to ethnic group and the form of NSCL/P. This study analyzed the association between the BMP4 gene polymorphisms rs762642, rs17563, and rs10130587 with the risk of cleft lip only (CLO), cleft palate only (CPO), and cleft lip with palate (CLP) in a population from South China.MethodsThis case-control study included 165 patients with NSCL/P (53 patients with CPO, 52 with CLO, and 60 with CLP) and 52 healthy volunteers. Peripheral blood samples were collected from all subjects to genotype the rs762642, rs17563, and rs10130587 polymorphisms by direct sequencing. Genotype and allelic frequencies of these polymorphisms were compared between healthy volunteers and patients with various forms of NSCL/P.ResultsThe genotype and allelic frequencies of rs762642 differed significantly between subgroups (CPO and CLP) and normal controls, whereas a significant difference was observed only in the CLO subgroup for the rs17563 polymorphism and in the CLO and CLP groups for the rs10130587 polymorphism. In addition, we identified a novel association of a BMP4 gene polymorphism, which was in linkage disequilibrium with the rs10130587 polymorphism, with CLO and CLP.ConclusionThe BMP4 gene polymorphisms rs762642, rs17563, and rs10130587 exhibit different associations with different forms of NSCL/P, suggesting that different forms of NSCL/P may have different etiologies.     

VAX1 gene associated non-syndromic cleft lip with or without palate in Western Han Chinese

ObjectiveNon-syndromic cleft lip with or without palate (NSCL/P) is one of the most common human birth defects, it results from multiple genetic and environmental risk factors. Recently, GWA studies identified associations between NSCL/P and two genetic risk loci, rs7078160 and rs4752028, at VAX1.DesignCurrently, we tried to investigate the roles of the two loci among 302 NSCL/P trios (129 non-syndromic cleft lip only (NSCLO) trios and 173 non-syndromic cleft lip and cleft palate (NSCLP) trios) from Western Han Chinese. The two SNPs were genotyped by SNPscan method; Hardy–Weinberg equilibrium test, allelic TDT and parent-of-origin effect were performed by PLINK software, and genotypic TDT and haplotype by FBAT software.ResultsAllelic TDT analysis revealed allele A at rs7078160 was over-transmitted among NSCL/P group (P = 0.0086, OR_transmission = 1.36, 95%CI: 1.08–1.72). Parent-of-origin effect analysis revealed a paternal special over-transmission of allele A at rs708260 in NSCL/P group (P = 0.0079). Haplotype AC of rs7078160-rs4752028 was significant over-transmitted in the NSCL/P group.ConclusionsOur study firstly confirmed that allele A at rs7078160 at VAX1 gene was a risk factor for NSCL/P in Western Han Chinese population.     

宁夏地区非综合征型唇腭裂环境因素的研究

目的 探讨宁夏地区非综合征型唇腭裂（NSCL/P）发病相关环境因素。方法 采用病例对照研究方法纳入NSCL/P患者453例,正常新生儿452例。对研究对象进行问卷调查,利用SPSS 16.0统计软件对数据进行卡方检验和Logistic回归分析。结果 NSCL/P患病类型构成比为唇裂︰唇裂合并腭裂︰腭裂=1︰2.02︰1.51。Logistic回归分析显示妊娠期发生异常、妊娠期感染、流产史、孕前孕中服用药物、饮茶、吸烟、饮酒、居住地附近工厂为危险因素（P<0.05）。单胎、早孕反应、食用豆制品食物、水果为保护因素（P<0.05）。结论 加强母亲孕期饮食均衡,避免感染、流产、服用药物以及不良生活习惯对降低NSCL/P的发生具有重要意义。     

中国西部人群IRF6基因SNPs与非综合征型唇腭裂相关性的研究

目的:探讨干扰素调节因子6 (IRF6) 基因rs2013162 和 rs2235375位点单核苷酸多态性(SNPs)与非综合征型唇腭裂的相关性.方法:收集病例组非综合征型唇腭裂患儿332 例,患者父亲243 例,患者母亲289 例,完整的核心家庭206个.对照组收集正常新生儿174 例.采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法检测IRF6基因这2 个多态位点基因型,进行病例对照和传递不平衡(TDT)分析.结果:在中国西部人群中,与正常对照组比较,唇腭裂组rs2235375位点的基因型和等位基因的频率存在统计学差异(均P<0.01).运用传递不平衡研究发现IRF6基因rs2235375位点的G等位基因在唇腭裂患者中存在过传递(P<0.01).有5 种单倍型组合显示有传递不平衡.结论:在中国西部人群中IRF6基因多态性与非综合征型唇腭裂的发生存在强的相关性.     

No evidence for a role of CRISPLD2 in non-syndromic cleft lip with or without cleft palate in an Italian population

Non-syndromic cleft lip with or without cleft palate (NSCLP) is a malformation with variable phenotypes, resulting from a mixture of genetic and environmental factors. Some studies have supported a role for the 16q24 region and its candidate gene, CRISPLD2, in clefting. A replication study is necessary to confirm these findings. The aim of the present study was to test, by genetic linkage and association analyses, whether the candidate gene, CRISPLD2, represents a risk factor for NSCLP. The analysis of 39 multigenerational families provided formal exclusion of a linkage between NSCLP and the CRISPLD2 locus under different genetic models and non-parametric analyses. The family-based study of 239 unrelated probands and their parents revealed no association between any particular allele or haplotype and NSCLP. Therefore, the present investigation did not support the hypothesis of the involvement of CRISPLD2 in NSCLP malformation, at least with regard to the Italian population.     

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目的    研究亚甲基四氢叶酸还原酶（MTHFR）基因 C677T多态性与山东地区非综合征性唇腭裂（NSCL／P）的相关性。方法    于2008 年 9月在山东省优生技术重点实验室采用聚合酶链反应-限制性片段长度多态性（ PCR-RFLP）分析,对2006 年8月至2008年8月曾在齐鲁医院治疗的来自山东地区NSCL／P患儿家庭34户和健康查体的正常儿童家庭46户的家庭成员MTHFR基因的C677T基因型进行检测。结果    携带T等位基因的父母,其子代患NSCL／P的危险性是不携带T等位基因父母的子代的2.420倍;母子都是TT突变纯合子,子代患NSCL／P的危险性是母子为非TT纯合子的4.162倍;子代是TT突变纯合子患NSCL／P的危险性是非TT纯合子的3.812倍。结论    山东地区NSCL／P与MTHFR基因 C677T的多态性相关,与父母的基因型存在联系;T基因在母子组合的研究中有统计学意义,父母传递给子代的T等位基因对后代的患病有重要作用。     

非综合征型唇腭裂与MTHFR基因多态性的相关性研究

目的：研究亚甲基四氢叶酸还原酶（methylenetetrahydrofolate reductase,MTHFR）基因位点C677T和A1298C与中国江苏地区汉族人群非综合征型唇腭裂（（nonsyndromic cleft lip with or without cleft palate,NSCL/P）发生的相关性。方法：采用聚合酶链反应-限制性片段长度多态性检测法对200例NSCL/P患者和213例健康人进行基因型检测。结果：MTHFR C677T对照组与病例组在基因型分布无统计学差异（P〉0.05）,TT基因型和携带T等位基因儿童罹患NSCL/P的风险分别是CC基因型儿童的1.84倍及1.57倍。进一步分层分析发现TT基因型和CT基因型能分别显著增加儿童唇裂伴或不伴腭裂和单纯性唇裂的发病风险。MTHFR A1298C病例组和对照组在基因型频率和等位基因频率有统计学差异（P〈0.05）,AC基因型和携带C等位基因的儿童罹患NSCL/P的风险分别比AA基因型儿童降低49%及43%。分层分析中,AC基因型和携带C等位基因可降低罹患唇裂伴腭裂及唇裂伴或不伴腭裂的风险。结论：MTHFR C677T可能为中国江苏地区汉族儿童NSCL/P的危险因素,而MTHFR A1298C有可能是NSCL/P发生的保护因素。     

Investigation of MYH14 as a candidate gene in cleft lip with or without cleft palate

Clefts of the orofacial region are among the most common facial defects and are caused by abnormal facial development during gestation. Cleft lip with or without cleft palate (CL/P) is a birth defect with a complex etiology resulting from a mixture of genetic and environmental factors. In the present study we considered myosin 14 ( MYH14 ) as a candidate gene for CL/P. This gene codes for the heavy chain of non-muscle myosin IIC (NMMHC-IIC), maps in the OFC3 region, and shares significant homology with myosin 9, a gene that our group has recently seen to be involved in CL/P. A linkage disequilibrium investigation was conducted with six single nucleotide polymorphisms in MYH14 and a sample of 239 CL/P nonsyndromic patients and their parents. Our family-based investigation provided no evidence of association between MYH14 and CL/P alleles. These data do not support the involvement of MYH14 in CL/P among the Italian population.     

中国人群非综合征性唇腭裂患者IRF6基因突变检测

目的 探讨干扰素调节因子6(interferon regulatory factor 6, IRF6) 在非综合征性唇腭裂(non-sydromic cleft lip and/or cleft palate,NSCL/P)患者中的突变情况。方法：收集119例NSCL/P患者及288名健康人对照样本的外周血血样并提取DNA。在IRF6基因的全部外显子分别设计引物,PCR扩增其序列,通过测序找出IRF6基因突变,并将这些突变在对照样本中进行验证。结果：共发现5种在正常人中没有的突变,其中4种是新发现的突变。结论：IRF6基因突变在中国人群中参与了非综合征唇腭裂疾病的发生。     

非综合征唇裂伴或不伴腭裂对口腔健康相关生活质量的影响

目的 探讨非综合征唇裂伴或不伴腭裂对口腔健康相关生活质量的影响。方法 回顾分析2017年1月—2019年6月山东省菏泽市立医院收治的非综合征唇裂伴或不伴腭裂90例患儿的临床资料,其中唇裂不伴腭裂52例（唇裂不伴腭裂组）,唇裂伴腭裂38例（唇裂伴腭裂组）;选择同期我院口腔科常规口腔检查、口腔健康的40例儿童作为对照组,所有入组者均进行口腔健康检查。应用问卷调查法评估口腔健康对日常生活的影响,采用SPSS 22.0软件包对口腔健康与日常生活质量的相关性进行Spearman相关分析。结果 唇裂不伴腭裂组与唇裂伴腭裂组在龋失补指数（DMFT）、功能牙数目（TH）、咬合牙对、龋齿牙数、LOA≥6 mm牙数间差异具有统计学意义（P<0.05）;唇裂不伴腭裂组与唇裂伴腭裂组的DMFT、TH、咬合牙对、龋齿牙数、LOA≥6 mm牙数均显著高于对照组（P<0.05）;唇裂伴腭裂组OIDP量表各条目得分、OIDP总分显著高于唇裂不伴腭裂组（P<0.05）;唇裂不伴腭裂组OIDP量表各条目得分、OIDP总分显著高于对照组（P<0.05）。采用Spearman法分析临床口腔健康指数与OIDP分数的相关关系,唇裂不伴腭裂组与唇裂伴腭裂组DMFT、MT与根龋牙数、LOA≥6 mm牙数与OIDP分数呈负相关;咬合牙对与OIDP分数呈正相关（P<0.05）。结论 非综合征唇裂伴或不伴腭裂的口腔健康对日常生活质量均有不同程度影响。     

The 19-bp deletion polymorphism of dihydrofolate reductase (DHFR) and nonsyndromic cleft lip with or without cleft palate: evidence for a protective role

Objective

Nonsyndromic cleft lip with or without cleft palate (NS-CL/P) are among the most common congenital birth defects worldwide. Several lines of evidence point to the involvement of folate, as well as folate metabolizing enzymes in risk reduction of orofacial clefts. Dihydrofolate reductase (DHFR) enzyme participates in the metabolic cycle of folate and has a crucial role in DNA synthesis, a fundamental feature of gestation and development. A functional polymorphic 19-bp deletion within intron-1 of DHFR has been associated with the risk of common congenital malformations. The present study aimed to evaluate the possible association between DHFR 19-bp deletion polymorphism and susceptibility to NS-CL/P in an Iranian population.

Material and Methods

The current study recruited 100 NS-CL/P patients and 100 healthy controls. DHFR 19-bp deletion was determined using an allele specific-PCR method.

Results

We observed the DHFR 19-bp homozygous deletion genotype (D/D) vs. homozygous wild genotype (WW) was more frequent in controls than in NS-CL/P patients (25% vs. 13%), being associated with a reduced risk of NS-CL/P in both codominant (OR=0.33, P=0.027) and recessive (OR=0.45, P=0.046) tested inheritance models. We also stratified the cleft patients and reanalyzed the data. The association trend for CL+CL/P group compared to the controls revealed that the DD genotype in both codominant (OR=0.30, P=0.032) and recessive models (OR=0.35, P=0.031) was associated with a reduced risk of CL+CL/P.

Conclusions

Our results for the first time suggested the DHFR 19-bp D/D genotype may confer a reduced risk of NS-CL/P and might act as a protective factor against NS-CL/P in the Iranian subjects.