孤儿核受体SHP敲减抑制BMP9诱导C3H10T1/2细胞的成骨分化  

Knockdown of Nuclear Receptor SHP Suppresses BMP9-induced Osteogenic Differentiation of C3H10T1 /2 Cells

作　　者：董谦[1] 李丽[1] 王玉凤[1] 冯巧灵[1] 吉彩霞[1] 刘晓骅 罗进勇[1] 

机构地区：[1]重庆医科大学检验医学院临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国生物化学与分子生物学报》2015年第2期161-167,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金项目(No.81272006)~~

摘　　要：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.The orphan nuclear receptor small hetero-dimer partner（ SHP） is a unique member of the nuclear receptor family,which contains a LXXLL and a ligand-binding domain,but lacks the typical domain for DNA binding. SHP regulates a variety of cellular events, such as cell proliferation,differentiation and metabolism,by interacting with various transcription factors. Whether SHP plays a role in BMP9-induced osteogenic differentiation has not yet been elucidated. In our study, SHP was demonstrated to involve in the differentiation of BMP9-induced C3H10T1 /2 cells into osteoblasts. SHP expression was up-regulated by BMP9 treatments as determined by RT-PCR and Western blot. Small interfering RNA was used to knock down the expression of SHP gene in C3H10T1 /2 cells,and reduced the expression of osteogenic-related genes Runx2,Id1,Id2 and CTGF,which were known to respond to BMP9 for up-regulations. The alkaline phosphatase（ ALP） activity / staining and Alizarin red staining showed that the knockdown of SHP expression inhibited BMP9-mediated osteogenic differentiation. The inhibition could be partially rescued by BMP9 overexpression. These results suggested that SHP was a required factor in BMP9-induced osteogenic differentiation in C3H10 1 /2 cells.

关 键 词：孤儿核受体SHP 重组腺病毒 小干扰RNA 骨形态发生蛋白9 C3H10T1/2细胞 

分 类 号：R34[医药卫生—基础医学] Q2[生物学—细胞生物学]