Inhibition of Ras-Mediated Cell Proliferation by Benzyloxybenzaldehyde

Abstract

Excessive proliferation of vascular smooth muscle cells (VSMC) in the intima is an important etiologic factor in vascular proliferative disorders such as atherosclerosis and restenosis after balloon angioplasty. Therefore, control of VSMC growth may be a suitable therapeutic intervention in vascular proliferative disorders. In the present work, we have studied the 2-benzyloxybenzaldehyde (CCY1a)-mediated antiproliferative effect and its mechanisms of action. CCY1a inhibited serum-induced VSMC proliferation in a concentration-dependent manner, as demonstrated using [³H]thymidine incorporation and MTT assays; the IC₅₀ values were calculated to be 7.0 × 10^–6 and 1.2 × 10^–5M, respectively. Furthermore, it also significantly suppressed serum-induced progression of the cell cycle, as shown by flow cytometric analysis. CCY1a as well as PD98059 almost completely abolished serum-induced activation of p42/44 mitogen-activated protein kinase (MAPK), the downstream effectors of c-fos and c-jun mRNA expression and activator protein-1 (AP-1) DNA binding activity, suggesting the central roles of these signaling cascades. Interestingly, CCY1a also effectively blocked serum-induced IĸB-α phosphorylation, IĸB-α degradation and nuclear factor-ĸB (NF-ĸB) binding activity. Based on these observations, we examined the effect of CCY1a on serum-mediated Ras activity, an upstream regulator of the above signaling events. The data demonstrated a marked inhibition of Ras activation by CCY1a. We conclude that CCY1a blocks cell proliferation via inhibition of the upstream effector of Ras and downstream events, including p42/44 MAPK activation and c-fos and c-jun mRNA expression, as well as NF-ĸB and AP-1 DNA binding activities.

CCY-1a-E2 induces G2/M phase arrest and apoptotic cell death in HL-60 leukemia cells through cyclin-dependent kinase 1 signaling and the mitochondria-dependent caspase pathway

“…In addition, 2-benzyloxybenzaldehyde can block Ca 2+ entry and suppress formyl peptide-stimulated increase in intracellular Ca 2+ in neutrophils (28). Pan et al (29) demonstrated that 2-benzyloxybenzaldehyde modulates vascular smooth muscle cell proliferation by blocking the Ras/p42/44 MAPK pathway and inhibiting NF-κB and AP-1 DNA binding activities. Our previous study showed that CCY-1a-E2 reduced the percentage of viable murine leukemia WEHI-3 cells, and the IC 50 value of CCY-1a-E2 was 5 µM for a 24-h treatment (19).…”

In vivo evaluation of the synthesized novel 2-benzyloxybenzaldehyde analog CCY-1a-E2 for the treatment of leukemia in the BALB/c mouse WEHI-3 allograft model

“…The results showed that 2-benzyloxybenzaldehyde exhibited antiproliferative effects on the vascular smooth muscle cells through the inhibition of the Ras/MAPK signal pathway and its downstream effectors (9). It has also been demonstrated that 2-benzyloxybenzaldehyde inhibits superoxide anion generation through the suppression of Akt and PLD activation in rat neutrophils (10,11).…”

The indazole derivative YD-3 inhibits thrombin-induced vascular smooth muscle cell proliferation and attenuates intimal thickening after balloon injury

“…Cultured VSMCs were lysed in a standard lyses buffer. The cell lysate was centrifuged and the supernatant was used for the Western blotting analysis as described previously (19). ERK1/2 and phosphorylated ERK1/2 antibodies (Santa Cruz Biotechnology) diluted 1 : 1000 in phosphate-buffered saline (PBS), were used as primary antibodies and were applied for overnight incubation at 4°C.…”