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Identification of Gli1‐interacting proteins during simvastatin‐stimulated osteogenic differentiation of bone marrow mesenchymal stem cells
Author(s) -
Chi Bojing,
Fan Xinhao,
Li Zhengxiao,
Liu Guangyuan,
Zhang Guobin,
Xu Hong,
Li Zhiguo,
Lian Qiangqiang,
Xing Lei,
Tian Faming
Publication year - 2019
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.29221
Subject(s) - gli1 , simvastatin , cyclopamine , hedgehog signaling pathway , microbiology and biotechnology , chemistry , signal transduction , osteocalcin , indian hedgehog , mesenchymal stem cell , mapk/erk pathway , runx2 , biology , alkaline phosphatase , biochemistry , endocrinology , enzyme
Simvastatin has been shown to promote osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Our study aimed to illuminate the underlying mechanism, with a specific focus on the role of Hedgehog signaling in this process. BMSCs cultured with or without 10 −7 mol/L simvastatin were subjected to evaluation of osteogenic differentiation capacity. Osteogenic markers such as type 1 collagen (COL1) and osteocalcin (OCN), as well as key molecules of Hedgehog signaling molecules, were examined by Western blot and real‐time polymerase chain reaction (PCR). Co‐immunoprecipitation and mass spectrometry assays were applied to screen for Gli1‐interacting proteins. Cyclopamine (Cpn) was used as a Hedgehog signaling inhibitor. Our results indicated that simvastatin increased alkaline phosphatase (ALP) activity; mineralization of extracellular matrix; mRNA expression of ALP, COL1, and OCN; and expression and nuclear translocation of Gli1. Contrasting effects were observed in Cpn‐exposed groups, but were partially rescued by the simvastatin treatment. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses indicated that Gli1‐interacting proteins were primarily associated with mitogen‐activated protein kinase (MAPK) ( P = 7.04E −04 ), hippo, insulin, and glucagon signaling. Further, hub genes identified by protein‐protein interaction network analysis included Gli1‐interacting proteins such as Ppp2r1a, Rac1, Etf1, and XPO1/CRM1. In summary, the current study showed that the mechanism by which simvastatin stimulates osteogenic differentiation of BMSCs involves activation of Hedgehog signaling, as indicated by interactions with Gli1 and, most notably, the MAPK signaling pathway.