摘要
目的研究Runx3敲减对BMP9诱导的小鼠胚胎成纤维细胞iMEFs成骨分化的作用。方法 BMP9腺病毒感染iMEFs,用Western blot检测内源性Runx3蛋白的表达。Runx3干扰腺病毒ad-siRunx3和BMP9条件培养基共同处理iMEFs,用碱性磷酸(ALP)染色和活性定量测定检测成骨早期指标ALP;用茜素红S染色检测成骨晚期指标钙盐沉积;用RT-PCR检测成骨相关基因Id1、Id2、Id3和Runx2,用Western blot检测成骨相关蛋白Dlx5;用SBE荧光素酶报告质粒检测smad1/5/8的转录活性。结果 BMP9可以使Runx3表达降低(P<0.05);干扰Runx3可以抑制早期成骨指标ALP活性(P<0.05)和晚期成骨指标钙盐沉积;ad-siRunx3抑制BMP9诱导的成骨相关转录因子Id1、Id2、Id3和Runx2基因的表达(P<0.05)及DLX5蛋白的表达(P<0.05)。结论敲减Runx3可以抑制BMP9诱导小鼠胚胎成纤维细胞向成骨分化。
Objective To analyse the effect of knocking down Runx3 on BMP9-induced osteogenic differenciation of iMEFs. Methods iMEFs were infected with ad-BMP9 and then the expression of endogenous Runx3 was determined by RT-PCR and Western blot. iMEFs were treated with ad-siRunx3 and BMP9,then the alkaline phosphatase（ ALP） activity was detected by quantitative and staining assay; calcium deposition was detected by Alizarin Red S staining; the gene expression levels of Id1,Id2,Id3 and Runx2 were determined by RT-PCR and the protein expression level of DLX5 was determined by Western blot. Results BMP9 inhibited the expression of Runx3. ad-siRunx3 can inhibit the expression of ALP,which is an early osteogenic marker,and the expression of calcium deposition,which is late osteogenic marker. ad-siRunx3 can inhibit the expression of Id1,Id2,Id3,Runx2 and DLX5 induced by BMP9（ P 〈 0. 05）. Conclusions Knocking down Runx3 can inhibit the BMP9-induced osteogenic differentiation of iMEFs.
出处
《基础医学与临床》
CSCD
2016年第2期173-178,共6页
Basic and Clinical Medicine
基金
国家自然科学基金(81272006)
重庆市基础与前沿研究计划(cstc2013jcyj A10061)
