Effect Of MS-275on RECK Gene Expression In Osteosarcoma Cells

ObjectiveTo observe the effect of histone deacetylase inhibitors Entinostat (MS-275) on RECKgene expression in osteosarcoma cells.MethodsOsteosarcoma cell line MG-63was cultured in vitro and divided into control groupand different concentrations of MS-275groups. For the control group, cells weretreated by0.1%DMSO；For the MS-275groups, cells were treated by0.1,1.0,3.0μmol/L MS-275for24~72h.The multiplication of MG-63cells were detected byWST-1assay. Total cellular RNA and protein was extracted, and expression of RECKmRNA and protein were detected by RT-PCR. Meanwhile, enzyme activity of Matrixmetalloproteinase-9(MMP-9) was detected by foerster resonance energy transferassay.Results1. WST-1results showed that0.1~3umol/L MS-275significantly inhibited theproliferation of MG-63cells, and the inhibiting rate reached to46%when3μmol/LMS-275was administrated.2. RT-PCR results showed, RECK gene was lowly expressed in MG-63cells. Aftertreatment of0.1~3μmol/L MS-275, the RECK mRNA increased. Western blot resultsalso showed, different concentration MS-275could increase the expression of RECKprotein in MG-63cells.3. Enzyme activity results showed, MMP-9Enzyme activity level was high in thesupernatant of MG-63cells. After0.1~3μmol/L MS-275treatment, enzyme activity significantly decreased.3μmol/L MS-275could decrease the enzyme activity rate to43%.ConclusionMS-275can increase the expression of mRNA and protein of RECK, and then affectthe enzyme activity of MMP-9to preventing tumors.