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孤儿核受体SHP(small heterodimer partner)是核受体超家族中的一员,具有LXXLL模体及配体结合域,但无经典的DNA结合域.它可与多种转录因子结合,调节细胞的增殖、分化和代谢等生物学过程.但目前关于SHP在BMP9诱导成骨分化中的确切作用却尚不清楚.本研究证明,SHP参与BMP9诱导的C3H10T1/2细胞成骨分化.RT-PCR结合Western印迹方法检测蛋白揭示,异位表达BMP9上调了SHP在C3H10T1/2细胞中的表达.小干扰RNA敲减SHP基因在C3H10T1/2细胞的表达下调了成骨相关基因Runx2、Id1、Id2及CTGF的表达,而过表达BMP9则可上调这些基因的表达.碱性磷酸酶(ALP)活性测定/染色及茜素红染色显示,敲减核受体SHP基因可抑制BMP9的成骨分化作用,而过表达BMP9可部分消除SHP敲减导致的成骨抑制作用.上述结果提示,核受体SHP为BMP9诱导的C3H10T1/2细胞成骨分化所必需.究竟BMP9如何上调SHP基因表达,以及SHP究竟通过何种机制上调BMP9下游成骨分化相关基因的表达尚待进一步研究.
Orphan nuclear receptor SHP (small heterodimer partner) is a member of the nuclear receptor superfamily, with LXXLL motif and ligand binding domain, but no classical DNA binding domain. It can bind a variety of transcription factors, regulating cells Differentiation, metabolism and other biological processes.However, the exact role of SHP in BMP9-induced osteogenic differentiation remains unclear.This study demonstrates that SHP is involved in BMP9-induced osteogenic differentiation of C3H10T1 / 2 cells.RT-PCR Detection of protein by Western blotting revealed that ectopic expression of BMP9 upregulated the expression of SHP in C3H10T1 / 2 cells, and knockdown of SHP gene in C3H10T1 / 2 cells downregulated the expression of Runx2, Id1, Id2 and CTGF expression, and overexpression of BMP9 upregulated the expression of these genes.Alkaline phosphatase (ALP) activity assay / staining and alizarin red staining showed that knockdown of nuclear receptor SHP gene can inhibit BMP9 osteogenic differentiation, Overexpression of BMP9 partially abolished the osteogenic effect induced by SHP knockdown.The above results suggest that nuclear receptor SHP is necessary for BMP9-induced osteogenic differentiation of C3H10T1 / 2 cells.However, how BMP9 up-regulates SHP gene expression, and whether SHP Through what mechanism Expression of transferred further downstream BMP9 osteogenic differentiation related genes yet.