Noggin基因沉默对BMP和Wnt信号通路表达的影响

Effect of Noggin silencing on the BMP and Wnt signaling pathways

目的检测分析Noggin基因沉默对毛囊发育中BMP和Wnt信号通路的影响。方法采用实时荧光定量PCR和western blot技术对Noggin基因沉默的MC3T3-E1稳转细胞系中BMP-2、BMP-4、BMPR-IA、BMP-6、BMP-7、LEF-1、β-catenin的表达情况进行检测分析。结果实时荧光定量PCR结果显示,BMP信号通路中的五个基因的表达都受到Noggin基因沉默的显著的影响,其中BMP-2(P<0.001)、BMP-4(P<0.01)、BMP-6(P<0.001)、BMP-7(P<0.001)表达量均升高;BMPR-IA(P<0.01)表达量降低。同时Wnt信号通路中的两个基因LEF-1(P<0.001)、β-catenin(P<0.001)的表达也都显著降低。Western blot结果显示,两条信号通路中这几种蛋白的表达也都受到影响,其中显著升高的有BMP-2(P<0.05)、BMP-4(P<0.05)、BMP-6(P<0.05)和BMP-7(P<0.05);显著降低的是β-catenin(P<0.05)、BMPR-IA(P<0.01)和LEF-1(P<0.001)。结论在体外Noggin基因对BMP信号通路可能存在反馈性抑制机制,而对Wnt信号通路存在反馈性激活机制,为下一步探究在体内Noggin基因对BMP和Wnt信号通路表达的作用提供一定的依据。

Objective To analyze the effect of Noggin silencing on the BMP and Wnt signaling pathways in hair follicle development. Methods The expression of BMP-2,BMP-4,BMPR-IA,BMP-6,BMP-7,LEF-1 and β-catenin in Noggin silencing MC3T3-E1 stable cell line was detected by RT-PCR and western blot. Results RT-PCR results showed that the expressions of five genes in BMP signaling pathway were all significantly influenced by Noggin silencing,the expressions of BMP-2（ P〈0. 001）,BMP-4（ P〈0. 01）,BMP-6（ P〈0. 001） and BMP-7（ P〈0. 001） were all increased and the expression of BMPR-IA（ P〈0. 01） was decreased. While the expressions of the two genes LEF-1（ P〈0. 001）and β-catenin（ P〈0. 001） in Wnt signaling pathway were significantly decreased. Western blot results showed that the expressions of these proteins in the two signaling pathways were also affected. The expressions of BMP-2（ P〈0. 05）,BMP-4（ P〈0. 05）,BMP-6（ P〈0. 05） and BMP-7（ P〈0. 05） were all increased,while the expressions of BMPR-IA（ P〈0. 05）,LEF-1（ P〈0. 01） and β-catenin（ P〈0. 001） were decreased. Conclusions There may be a negative feedback regulation of Noggin on the BMP signaling pathway in vitro,but a positive feedback regulation on the Wnt signaling pathway in vitro. It provides certain evidence for studies on the effect of Noggin gene on BMP and Wnt signaling pathways in vivo.There may be an interaction between hair follicle development-related signaling pathways,which still needs further experiments to prove.