尘肺病易感基因SNPs多位点图谱的构建

Construction of a multilocus map of single nucleotide polymorphisms in pneumoconiosis susceptibility genes

目的构建尘肺病易感基因单核苷酸多态性(single nucleotide polymorphisms,SNPs)多位点检测图谱。方法利用限制性片段长度多态性(restriction fragment length polymorphism,RFLP)技术,对已报道的易感基因SNPs位点进行整合,并根据基因位点设计不同的扩增片段和酶切片段,构建涵盖多个不同易感基因SNPs位点组成的多位点图谱,通过琼脂糖凝胶电泳的多个长短不一的条带快速实现基因分型;采用陕北某矿区煤工尘肺(coal workers′pneumoconiosis,CWP)和接尘人群队列进行检测,通过遗传分化指数(fixation index,Fst)分析易感基因不同多态位点在队列遗传分化情况。结果成功构建5个易感基因IL-1βrs16944 T>C、IL-6 rs1800796 G>C、IL-8 rs4073 A>T、HSP70-1 rs562047 C>G、TGF-βrs1800469 T>C的SNPs图谱,可快速有效鉴别SNPs的基因型;Fst分析表明IL-1βrs16944、TGF-βrs1800469的Fst为>0.05~0.15,即发生中等程度遗传分化。结论该易感基因SNPs多位点图谱可以有效实现基因分型,易感位点rs16944和rs1800469更易受到疾病选择压力的影响,与尘肺病的易感风险有关。

Objective To construct a single nucleotide polymorphisms(SNPs)multilocus map of pneumoconiosis susceptibility genes.Methods Based on RFLP technology,we integrated the reported SNPs of susceptibility genes,and designed different amplification fragments and digestion fragments which cover several different susceptibility gene locus.Then the multiple bands of different lengths obtained by agarose gel electrophoresis were used to identify the genetic typing rapidly.In addition,the cohort of coal workers′pneumoconiosis(CWP)and dust receiving population in a mining area in northern Shaanxi was used for detection,and the genetic differentiation of different polymorphic loci of susceptible genes in the cohort was analyzed by fixation index(Fst).Results The SNPs of five susceptible genes were successfully constructed,which included IL-1βrs16944 T>C,IL-6 rs1800796 G>C,IL-8 rs4073 A>T,HSP70-1 rs562047 C>G,TGF-βrs1800469 T>C.These SNPs genotypes can be identified effectively.Fst analysis showed that the Fst of IL-1βrs16944 and TGF-βrs1800469 was>0.05-0.15,indicating that,moderate genetic differentiation occurred.Conclusions The susceptibility loci rs16944 and rs1800469 were more susceptible to disease selection pressure,which was related to the susceptibility risk of pneumoconiosis.