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Objective To investigate the effect of MS-275, an histone deacetylase ( HDAC ) inhibitor, on acute liver failure ( ALF ) induced by D-galactosamine and lipopolysaccharide in mice. Methods Thirty specific pathogen free (SPF) C57BL/6 male mice were randomly and equally divided into control, ALF model and MS-275 groups. ALF model was induced by D-galactosamine ( D-Gal ) and lipopolysaccharide (LPS), and the mice in MS-275 group received MS-275 (1 mg/kg) at 2 h before the induction of ALF.Serum and liver samples of mice were obtained at 24 h after ALF induction.The serum levels of ALT, AST, TBil and tumor necrosis factor-alpha ( TNF-α) , interferon γ( IFNγ) , interleukin ( IL )-1β, high mobility group box 1 ( HMGB1 ) were tested by biochemical methods or ELISA kit, respectively.The expression of HDAC1, HDAC3, acetylation of histone H3, H4, P65, acetylation and phosphorylation of P65 in liver were detected by Western blotting.The changes of histology in liver was detected by HE staining, and the translocation of P65 in liver was detected by immunohistochemistry. Comparison of variables among the groups was performed using t test.Results MS-275 inhibited the infiltration of inflammatory cells and improved the pathological changes of liver tissue.Compared with ALF group, serum ALT, AST, TBil levels were decreased in MS-275 group ( t =-22.215, -11.914 and-12.160, all P<0.05), but still higher than those in the control group (t=14.852, 11.692 and 8.333, all P<0.05); serum TNF-α, IFNγ, IL-1β, HMGB1 levels were also significantly decreased in MS-275 group (t=-7.926, -3.427, -2.475 and -5.920, all P<0.05), but TNF-αand IFNγwere still higher than those in the control group (t=5.541 and 5.514, all P<0.05).Compared with control group, the expression of class I HDAC in liver tissue was significantly decreased in MS-275 group ( t=-3.676 and-10.576, P<0.05), while the expressions of acetylation of histone H3, H4 and P65 were significantly increased (t=3.976, 5.559 and 4.588, all P<0.05).MS-275 inhibited the translocation of P65 from cytoplasm to the nucleus.Conclusion MS-275 can protect liver from acute failure in mice through enhancing the acetylation levels of non-histones.
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Objective The use of targeting therapy for the treatment of gastric glandular cancer has been a hot topic in recent years. This study aims to clarify that through what ways the histone deacetylase inhibitor MS-275 completes its selectively killing effect on gastric glandular cancer cell line SGC-7901. Methods SGC-7901 cells and GES-1 cells were respectively cultured for 24h, with(10-100) μmol/L concentrations of MS-275. (1) The survival rate of SGC-7901 cells, GES-1 cells and the normal cells were analyzed by WST-1; (2) The change of the mitochondrial membrane potential in SGC-7901 was estimated by flow cytometry;(3) The expression levels of p21, p27, p57, cyclinB1, cyclinD1 were determined by Western blot and PCR methods. Results (1) MS-275 could decrease the survival rate of SGC-7901 cells, the effect was significantly enhanced with the increasing of the concentration (P<0.05), but MS-275 showed no obvious effect on normal gastric mucosa epithelial cells GES-1; (2) MS-275 treatment could decreased the mitochondrial membrane potential of SGC-7901 cells (P<0.05); (3) MS-275 treatment could increase the relative contents of p21, p27, p57 genes and their protein and, at the same time, decrease the relative contents of CyclinB1 and CyclinD1 (P<0.05). Conclusion MS-275 treatment can selectively kill gastric glandular cancer cells SGC-7901 through several possible ways, such as inducing mitochondrial apoptosis and regulating the expression levels of cell cycle-related genes and proteins.
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PURPOSE: This study was conducted to identify the surface characteristics of titanium discs coated with MS275/PLGA by electrospray and which is effective to mesenchymal stem cell proliferation. MATERIALS AND METHODS: We used anodized surface coated with PLGA as a control group and anodized surface coated with MS275 0.5 microM, 1 microM, 1.5 microM as test groups. To examine that the coating particles are nanometer sized, FE-SEM was used and AFM was utilized to determine the difference of coating surface roughness. We checked the mesenchymal stem cell proliferation by using MTT assay on 1st, 4th, 7th days. RESULTS: There was no significant difference between control groups and test groups in AFM results (P>.05). In MTT assay results, mesenchymal stem cell proliferation was increased with time, at 7th day, cell viability on discs coated with 1.5 microM MS275 was significantly higher than control group (P<.05). As SEM showed, the number of cells on all discs was increased and the morphology of cell attachment was also wider and closer with time. CONCLUSION: Titanium surface coated with MS275/PLGA showed significantly higher cell proliferation and the more density of MS275 was dispersed on titanium discs, the faster cells grew.
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Cell Proliferation , Cell Survival , Lactic Acid , Mesenchymal Stem Cells , Polyglycolic Acid , TitaniumABSTRACT
Objective:To investigate the influence of combined application of histone deacetylase inhibitor(HDACi)MS-275 and 5-FU on the apoptosis and cycle arrest on human hepatoma cell line HepG2 and unclose the related mechanism.Methods:Divided all the cells into 4 groups:control group,MS-275 group,5-FU group and drug combination group.Flow cytometry(FCM)was used to examine the effects of 5-FU with MS-275 on the apoptosis and cell cycle of HepG2 cells.Bcl-2,Bax,CyclinD1 and P21 protein were determined by Western blot assay.Results:5-FU and MS-275 combination could inhibit HepG2 cell growth through G0-G1 arrest,and induce apoptosis,both time and dose dependently.The combination of two agents increased P21 protein levels and decreased Bcl-2,CyclinD1 protein levels.The levels of Bax protein were not changed.Conclusion:The combination of 5-FU and MS-275 can induce apoptosis and cell cycle arrest,the effect might be associated with down-regulating expression of Cyclin D1 and Bcl-2 protein and upregulating the expression of P21 protein.
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Objective Curcumin and MS-275,an inhibitor of histone deacetylase (HDAC),are promising anti-tumor agents. The aim of present study was to investigate the mechanisms of apoptosis induced by combined use of MS-275 in low dosage and curcumin in prostate cancer cell line DU145. Methods MTT assay was used to evaluate the lethal effect on DU145 cells by solitary use of MS-275 and or combined with curcumin. The changes in cell life cycle was detected by flow cytometry. The expressions of the survival signaling pathways were determined by Western blotting. Results Solitary application of MS-275 or curcumin may inhibit the growth of DU145 cells in a time and dose-dependent manner. The combination of MS-275 (1?mol/L) and curcumin (20?mol/L) exhibited obvious cytotoxic effect on cell viability,which was only 45.9% 48h after the combined treatment. Cell cycle assay showed that the combination of MS-275 and curcumin resulted in obvious appearance of sub-G1 phase in DU145 cells,implying that the cell apoptosis had been induced. The results of Western blotting showed that after treatment of MS-275 or curcumin singly,the phosphorylation level of Akt and ERK kinase declined slightly,however,when MS-275 and curcumin were used together,there appeared a prominent inhibitory effect on Akt and ERK kinase,indicated by a sharp decline of their phosphorylation level,and at the same time,the level of cleaved PARP,a hallmark of apoptosis,was increased in DU145 cells. Conclusion Combined use of MS-275 and curcumin may exert a synergistic cytotoxic effect on the viability of DU145 cells,and exhibit an inhibitory activity on Akt and ERK kinases to induce apoptosis.