中国唇腭裂患者Sonic hedgehog信号通路相关单核苷酸多态性的分析

目的 探讨Sonic hedgehog(Shh)信号通路相关的单核苷酸多态性(single-nucleotide polymorphism, SNP)与非综合征型唇腭裂(non-syndromic cleft lip and/or palate, NSCL/P) 之间的关联,并对唇腭裂疾病的致病风险因素进行探索。方法 收集197例个体的外周血(NSCL/P患者100例,健康对照97例),基于国际人类基因组单体型图计划中中国北京汉族人口数据,使用Haploview软件进行单倍体型分析和标签SNP选择。针对Shh 信号通路中的4个候选基因SHH、PTCH1、SMO和GLI2共选择了27个SNP。使用Sequenom质谱技术检测27个SNP在4个Shh信号通路中候选基因的基因型,并进行分析。结果 所选择的SNP基本涵盖了候选基因的潜在功能性SNP,其最小等位基因频率(minor allele frequency,MAF)>0.05:GLI2 73.5%, PTCH1 91.0%, SMO 100.0%, SHH 75.0%。发现位于SMO基因的SNP(rs12674259)和位于PTCH1基因的SNP(rs2066836)的基因型频率在NSCL/P病例组和对照组之间的差异有统计学意义。同时在4个候选基因所在的3条染色体(第2、7、9号染色体)中均发现了连锁不平衡,但在连锁不平衡单倍体型分析中,病例组和对照组之间的差异无统计学意义。结论 提示Shh信号通路参与NSCL/P的发生,其信号通路中关键基因的某些特殊SNP位点与唇腭裂相关,为NSCL/P的病因研究提供了新的探索方向,可能为NSCL/P的早期筛查与风险预测提供帮助。

Analysis of single-nucleotide polymorphism of Sonic hedgehog signaling pathway in non-syndromic cleft lip and/or palate in the Chinese population

Objective: To study the relationship between Sonic hedgehog (Shh) associated single-nucleotide polymorphism (SNP) and non-syndromic cleft lip and/or palate (NSCL/P), and to explore the risk factors of cleft lip and/or palate. Many studies suggest that the pathogenesis of NSCL/P could be related to genes that control early development, in which the Shh signaling pathway plays an important role.Methods: Peripheral blood was collected from 197 individuals (100 patients with NSCL/P and 97 healthy controls). Haploview software was used for haplotype analysis and Tag SNP were selected, based on the population data of Han Chinese in Beijing of the international human genome haplotype mapping project. A total of 27 SNP were selected for the 4 candidate genes of SHH, PTCH1, SMO and GLI2 in the Shh signaling pathway. The genotypes of 27 SNP were detected and analyzed by Sequenom mass spectrometry. The data were analyzed by chi-squared test and an unconditional Logistic regression model. Results: The selected SNP basically covered the potential functional SNP of the target genes, and its minimum allele frequency (MAF) was >0.05: GLI2 73.5%, PTCH1 91.0%, SMO 100.0%, and SHH 75.0%. It was found that the genotype frequency of SNP (rs12674259) located in SMO gene and SNP (rs2066836) located in PTCH1 gene were significantly different between the NSCL/P group and the control group. Linkage disequilibrium was also found on 3 chromosomes (chromosomes 2, 7 and 9) where the 4 candidate genes were located. However, in the analysis of linkage imbalance haplotype, there was no significant difference between the disease group and the control group.Conclusion: In China, NSCL/P is the most common congenital disease in orofacial region. However, as it is a multigenic disease and could be affected by multiple factors, such as the external environment, the etiology of NSCL/P has not been clearly defined. This study indicates that Shh signaling pathway is involved in the occurrence of NSCL/P, and some special SNP of key genes in this pathway are related to cleft lip and/or palate, which provides a new direction for the etiology research of NSCL/P and may provide help for the early screening and risk prediction of NSCL/P.