0.5-Gy X-ray irradiation induces reorganization of cytoskeleton and differentiation of osteoblasts

Osteoblasts are sensitive to ionizing radiation. The small GTPase RhoA and its effector Rho-associated protein kinase (ROCK) are critical to several cellular functions, including cytoskeleton reorganization, cell survival, and cell differentiation. However, whether the RhoA/ROCK signaling pathway is involved in the regulation of osteoblast cytoskeleton reorganization and differentiation induced by low-dose X-ray irradiation remains to be determined. The aim of the present study was to investigate the role of the RhoA/ROCK signaling pathway in mediating differentiation of osteoblasts and reorganization of the cytoskeleton under low-dose X-ray irradiation. Osteoblasts were pretreated with the ROCK kinase-specific inhibitor (Y-27632) before exposure to low-dose X-ray irradiation. The changes of F-actin in MC3T3 cells were observed at different time points following X-ray irradiation. Cell Counting Kit-8 assay, alkaline phosphatase activity, Alizarin red staining and western blotting were used to detect the proliferation and differentiation of osteoblasts after 0.5-Gy X-ray irradiation. In the present study, low-dose X-ray irradiation promoted the expression of genes associated with the cytoskeleton reorganization. Indeed, the results showed that, 0.5-Gy X-ray irradiation can induce reorganization of cytoskeleton and promote differentiation of osteoblasts through the RhoA/ROCK signaling pathway. Additionally, inhibiting ROCK activity blocked low-dose X-ray irradiation-induced LIMK2 phosphorylation, stress fiber formation and cell differentiation. Thus, these results demonstrated the excitatory effects of low-dose X-ray irradiation on MC3T3-E1 cells, including reorganization of the cytoskeleton and differentiation of osteoblasts.