Sphingosine-1-phosphate inhibits differentiation of C3H10T1/2 cells into adipocyte

Abstract

Mesenchymal stem cells (MSCs) can differentiate into a number of cell types, including adipocytes and osteoblasts. MSC differentiation into adipocytes inhibits osteogenic differentiation and vice versa. Therefore, understanding the mechanisms of MSC differentiation at the signaling level can lead to the development of novel therapeutic strategies toward tissue regeneration. Sphingosine-1-phosphate (S1P) is a signaling molecule that regulates many cellular responses, including cellular differentiation. However, the effects of S1P on MSC differentiation are largely unknown. The purpose of study was to investigate whether S1P drives MSCs toward either adipogenic or osteogenic differentiation, and if so, to clarify the underlying signaling mechanisms for such differentiation. We found that S1P inhibited adipogenic differentiation of C3H10T1/2 multipotent stem cells, while promoting their osteogenic differentiation. During adipogenic differentiation, S1P suppressed the cAMP accumulation in a Gi-protein-dependent manner. The Gi-dependent S1P signaling suppressed C/EBPβ expression, which is essential for adipogenic differentiation. Furthermore, S1P did not affect cAMP-independent adipogenic differentiation. These findings suggest that S1P suppresses cAMP accumulation, leading to inhibition of C/EBPβ expression, thereby resulting in decreased adipogenic differentiation of C3H10T1/2 cells. Thus, our findings provide novel molecular mechanisms as regards how S1P inhibits adipogenic differentiation of C3H10T1/2 cells, indicating a potential beneficial role for regeneration and repair of tissues.

Sphingosine‐1‐phosphate‐enhanced Wnt5a promotes osteogenic differentiation in C3H10T1/2 cells

“…It is generally believed that the induction of osteoblastogenesis of MSC leads to the inhibition of adipogenesis and vice versa (Cawthorn et al, ; Rahman et al, ). The current study and our recent study (Hashimoto et al, ) indicate that S1P may regulate both the promotion of osteoblastogenesis and the suppression of adipogenesis in MSC. Therefore, we speculate that S1P may be a suitable therapeutic agent for senile osteoporosis, which is characterized by both an increased number of adipocytes and a decreased number of osteoblasts.…”

“…Therefore, our experimental system may not be suitable for the observation of OC expression. However, our previous studies showed that S1P enhanced osteoblastmediated calcification in osteoblast-like MC3T3-E1 cells (Matsuzaki et al, 2013) and in C3H10T1/2 cells (Hashimoto et al, 2015). Consequently, understanding the role of S1P in MSC differentiation is important for facilitating bone regeneration.…”

“…We (Matsuzaki et al, 2013) previously reported that S1P enhances osteogenic differentiation of osteoblast-like cells. We also observed that S1P promotes the mRNA level of osteoblast differentiation markers in C3H10T1/2 pluripotent stem cells (Hashimoto et al, 2015). However, the mechanism of S1P-modulated mesenchymal stem cells (MSC) differentiation into osteoblasts remained unclear.…”

Sphingosine-1-phosphate/S1PR2-mediated signaling triggers Smad1/5/8 phosphorylation and thereby induces Runx2 expression in osteoblasts

“…Consistently, S1P/S1PR1 also regulates osteoclastogenesis [17]. However, we have previously demonstrated that a substantial amount of S1PR2 mRNA is expressed in osteoblast-like cells [10], C3H10T1/2 cells (mesenchymal stem cell-cell lines) [18]. Gi-independent S1P/S1PR2 signaling also activates RhoA/Rho kinase (ROCK) in smooth muscle cells and CHO cells [19,20].…”

The role of the progressive ankylosis protein (ANK) in adipogenic/osteogenic fate decision of precursor cells

“…Recent studies have suggested that S1P may play an important role in the regulation of osteogenesis. Specifically, these studies have shown that S1P promotes osteogenic differentiation of precursor cells and inhibits their adipogenic differentiation [47–49]. Future studies have to determine the exact mechanisms of how ANK affects osteogenic and adipogenic differentiation of precursor cells.…”