7-Dehydrocholesterol reductase regulated the palatal development by the sonic hedgehog-bone morphogenetic protein 2 signal pathway
10.3760/cma.j.issn.1002-0098.2014.12.007
- VernacularTitle:7-脱氢胆固醇还原酶基因沉默对体外培养腭突音猬基因-骨形成蛋白2信号通路的影响
- Author:
Daizun ZHANG
1
;
Yaoxiang XU
;
Wenlin XIAO
;
Cuizhu ZHUANG
Author Information
1. 青岛大学附属医院儿童口腔科
- Keywords:
Palate;
Cleft palate;
Gene silencing;
Maxillofacial development
- From:
Chinese Journal of Stomatology
2014;49(12):728-732
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of 7-dehydrocholesterol reductas(Dhcr-7) gene silencing on the palatal development by sonic hedgehog(Shh)-bone morphogenetic protein2(BMP-2) signal pathway in vitro.Methods A total of 60 pairs of palatal shelves fromgestation day(GD) 13.5 mouse embryos were divided into three groups(A,B,C) of 20 randomly.In group A(control),palatal shelves were cultured with medium containing no cholesterol.In group B(Dhcr-7-siRNA),palatal shelves were cultured without cholesterol medium but containing Dhcr-7 siRNA adenovirus.After 48h,the culture medium of groups A and B were changed with medium without cholesterol.In group C(cholesterol),palatal shelves were cultured without cholesterol medium but containing Dhcr-7 siRNA adenovirus.After 48h,the culture medium of group C was changed with medium containing 600 mg/L cholesterol.After 72h again,tissues dyeing and scanning electron microscope(SEM) technique were used to observe morphological changes of palates.Both RT-PCR and Western blottingtechniques were used to measure mRNA and protein expressions for Dhcr-7,Shh,and BMP-2,respectively.Results The tissues dyeing and SEM showedthat the palates fusedin groups A and Candthe palates did not fuse in group B eventually.The expression of both mRNA and proteins for Shh and BMP-2 in group B wasdecreased with the Dhcr-7 reduction.In group B,the mRNA and protein expression of Shh was separately 0.063±0.018 and 0.092±0.065;the mRNA and protein expression quantity of BMP-2 was separately 0.054±0.018 and 0.049±0.021.In group A,the mRNA and protein expression of Shh was separately 0.667±0.093 and 0.639±0.078;the mRNA and protein expression of BMP-2 was separately 0.591 ±0.043 and 0.569± 0.081.The difference of Shh and BMP-2 mRNA and protein expression between A and B group were statistically significant separately(P＜0.05).The expression of both mRNA and protein for Dhcr-7(0.074±0.034 and 0.075±0.028) did not changebasicallyin group C,compared with the Dhcr-7expression of mRNA and protein(0.083 ± 0.045;0.067± 0.065) in group B,the difference wasnot statistically significant(P＞0.05).In group C,the mRNA and protein expressionof Shh(0.649±0.085 and 0.608±0.092) and BMP-2(0.578±0.062 and 0.548±0.065) were significantly increased.The difference of Shh and BMP-2 mRNA and protein expression between B and C group were statistically significant separately(P＜ 0.05).Conclusions Dhcr-7 could influence the expression of Shh and BMP-2.Dhcr-7 reductase regulated the palatal development by the Shh-BMP-2 signal pathway.
