Shh与Wnt5a基因在Cornelia De Lange综合征中的表达及意义

Expression and significance of Shh and Wnt5a genes in Cornelia de Lange syndrome

目的通过研究Shh、Wnt5a基因在NIPBL^(+/-)胎鼠肢芽中的表达情况,探讨两基因与CdLS综合征的相关性。方法实验组取NIPBL^(+/-)胎鼠肢芽(n=36)和对照组取NIPBL^(+/+)胎鼠肢芽(n=36),分别于妊娠(E)10 d、E11 d、E12 d(n=12)取胎鼠肢芽组织,应用real-time PCR、Western blot技术分别检测Shh、Wnt5a基因转录及蛋白表达水平。结果 E10 d、E11 d、E12 d时,Shh、Wnt5a基因及其蛋白在两组胎鼠肢芽中均有表达,其表达水平在实验组与对照组比较均降低(P<0.01)。实验组和对照组胎鼠肢芽内Shh、Wnt5a基因及其蛋白表达水平在E10 d时均较少,E11 d时升高,随后E12 d时表达水平减少,且低于E10 d时的水平(P<0.01)。结论 Shh及Wnt5a基因与其相关蛋白水平表达存在一致性;CdLS综合征的发病可能与NIPBL基因低表达,从而抑制Shh及Wnt5a基因及其蛋白的表达相关。[中国当代儿科杂志,2019,21(5):485-490]

Objective To study the expression of Shh and Wnt5 a genes in the limb buds of NIPBL+/-fetal rats and the association of these two genes with Cornelia de Lange syndrome(CdLS).Methods A total of 72 NIPBL+/-fetal rats were divided into an experimental group and a control group,with 36 rats in each group.The limb buds were collected from 12 fetal rats each on embryonic days 10,11 and 12(E10,E11 and E12)respectively.Real-time PCR and Western blot were used to measure the mRNA and protein expression of Shh and Wnt5 a.Results The mRNA and protein expression of Shh and Wnt5 a was detected in the limb buds on E10,E11 and E12,and the experimental group had significantly lower expression than the control group(P<0.01).The mRNA and protein expression of Shh and Wnt5 a in limb buds was at a low level on E10,followed by an increase on E11 and a reduction on E12,and the expression on E12 was still lower than that on E10(P<0.01).Conclusions The mRNA and protein expression of Shh and Wnt5 a are consistent.The pathogenesis of CdLS may be associated with the low mRNA and protein expression of Shh and Wnt5 a inhibited by the low expression of NIPBL gene.[Chin J Contemp Pediatr,2019,21(5):485-490]