Role of vasodilator‑stimulated phosphoprotein in RANKL‑differentiated murine macrophage RAW264.7 cells: Modulation of NF‑κB, c‑Fos and NFATc1 transcription factors

Abstract

Vasodilator-stimulated phosphoprotein (VASP) is essential for osteoclast differentiation, and reduced VASP expression results in depressed osteoclast differentiation. Previously, we demonstrated the importance of VASP and Ras-related C3 botulinum toxin substrate 1 interactions in osteosarcoma cell migration and metastasis using Mg-63 and Saos2 cells. However, the molecular details of the functional role of VASP in cell motility and migration remain to be elucidated. The present study demonstrated that VASP affects the expression of αV-integrin, tartrate-resistant acid phosphatase (TRAP) and lamellipodia protrusion in RAW 264.7 murine macrophage cells. The RAW 264.7 mouse monocyte macrophage cell line was used as an osteoclast precursor. RAW 264.7 cells were treated with 50 ng/ml of receptor activator of nuclear factor κ-Β ligand (RANKL) in order to induce cell differentiation (osteoclastogenesis). Small interfering RNA (siRNA) was used to silence VASP, and RT-PCR and western blotting were used to determine the expression for genes and proteins, respectively. TRAP staining as a histochemical marker for osteoclast and fluorescent microscopy for lamellipodia protrusion was performed. RANKL treatment significantly increased the gene and protein expression of VASP, αV-integrin and TRAP in RAW 264.7 cells. Silencing of VASP significantly reduced the RANKL-induced expression of αV-integrin, TRAP and lamellipodia protrusion. In addition, knockdown of VASP attenuated RANKL-stimulated activation of NF-κB, c-Fos and nuclear factor of activated T cells cytoplasmic 1 transcription factors, and the phosphorylation of the p65 and IκBα. These results suggest the critical role of VASP in regulating osteoclast differentiation, which should be further explored in osteosarcoma research.

Vericiguat Modulates Osteoclast Differentiation and Bone Resorption via a Balance between VASP and NF-κB Pathways

“…To further expound the potential regulatory mechanism through which Vericiguat regulated the NF- κ B pathway, we found increasing expression of VASP induced by either Vericiguat only or RANKL in a dose-dependent manner. Furthermore, in vitro experiment suggested that VASP was essential to OC differentiation, consistent with a recent study from Hu et al [ 35 ]. Additionally, we found that Vericiguat only could also dually regulate the activation of NF- κ B.…”

“…The results of molecular modeling experiments confirmed our hypothesis that high-dose Vericiguat may directly disturb the activation of the NF- κ B signaling cascade. In fact, Flores-Costa et al reported that another sGC stimulator, praliciguat, could also reduce the phosphorylation of I κ B and NF- κ B to exert anti-inflammatory effect [ 35 ]. We deduced that Vericiguat may dominantly activate the sGC/VASP/I κ B- α /NF- κ B signaling pathway at low concentration.…”

Clinical Significance of Detection of Peripheral Blood VASP Level in Lung Cancer Patients

“…In recent years, studies have found that VASP is a cancer-promoting molecule [ 1 ]. The deletion of VASP can lead to the generation of tumor cells, and its expression level causes transformation of tumor cells [ 2 , 3 ]. Interestingly, VASP is highly expressed in lung adenocarcinoma tissues.…”

“…Now it is the leading cause of tumor. Despite the continuous progress in the diagnosis and treatment of lung cancer, no signi cant improvement was achieved in its prognosis and patients' survival time [1][2][3][4][5][6][7]. To make matters worse, the prognosis and survival rate of lung cancer noticeably decrease as it moves to more advanced stages [8][9][10].…”

N-Butanol Extract of Modified You-Gui-Yin Attenuates Osteoclastogenesis and Ameliorates Osteoporosis by Inhibiting RANKL-Mediated NF-κB Signaling

“…RANKL signaling is thought to be the major target of antiresorptive agents that inhibit bone loss and osteoclast activation. RANKL-induced osteoclast differentiation activates NF-kB and NFATc1 (42)(43)(44). Under normal conditions, NF-kB protein is stored as an inactive form in the cytoplasm (the p50/p65 heterodimer associated with repressive IkBa) (45).…”