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Imidazole-dione conjugate induces apoptosis and inhibits proliferation of osteosarcoma cells via activation of p65NFκB
Abstract
Purpose: To investigate the effect of imidazole-dione conjugate (IMC) on proliferation of MG63 osteosarcoma cells.
Methods: The effect of IMC on proliferation of MG63 osteosarcoma cells was determined using 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, while mRNA expressions of PTEN, FasL and FasR were assayed with real-time reverse transcription polymerase chain reaction (RT-PCR). Cell apoptosis was studied by flow cytometry. The protein expression level of IκBα was determined using western blotting.
Results: There were reductions in the proliferation of IMC-treated MG63 cells and Saos-2 cells at IMC dose of ≥ 4 μM (p < 0.05). Degree of proliferation of MG63 cells on exposure to 1, 2, 4, 6, 8 and 10 μM IMC was 99, 98, 76, 59, 34 and 21 %, respectively, relative to 100 % in untreated cultures. In MG63 cell cultures, treatment with 4, 6, 8 and 10 μM IMC led to 22, 39, 62 and 69 % apoptosis, respectively, when compared with 0.9 % apoptosis in control cell cultures (p < 0.05). Concentration-dependent increases were observed in PTEN, FasL and FasR mRNA in IMC-treated MG63 cells. Western blot assay showed a marked increase in the level of IκBα in MG63 cells following treatment with IMC. IMC treatment also caused a concentration-dependent increase in the expression of phospho-Ser536 p65NF-κB (p < 0.05).
Conclusion: IMC exerts inhibitory effect on the proliferation of MG63 cells via up-regulation of NF-κB phosphorylation. Thus, IMC may be useful as a therapeutic agent for osteosarcoma.