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

摘要:

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。

Abstract:

Objective To study the expression of Shh and Wnt5a 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 Wnt5a. Results The mRNA and protein expression of Shh and Wnt5a 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 Wnt5a 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 Wnt5a are consistent. The pathogenesis of CdLS may be associated with the low mRNA and protein expression of Shh and Wnt5a inhibited by the low expression of NIPBL gene.