Effects Of Histone Deacetylase Inhibitor Trichostatina A And Panobinostat On The Proliferation Of Hereditary Gingival Fibroblastoma Cell And The Mechanism

Objective:The aim of this study was to explore the influnces of Histone deacetylase inhibitor Trichostatina A and Panobinostat on cell proliferation of hereditary gingival fibroblastoma cells and to explore the underlying mechanisms.Methods:1.H-GMSCs were gently separated from HGF gingival tissues which were then induced into osteogenesis and adiponenesis differentiation to detect the multiple differentiate potentials.Flow assay was adopted to detect the surface maker of MSC.Osteogenesis and adipogenesis differentiation were observed by Alizarin Red staining and Oil-O Red staining.Mesenchymal stem cells isolated from healthy gingiva were performed as control.2.The effect of histone deacetylase inhibitor on the proliferation,apotosis and cell cycle of H-GMSCs and the molecular mechanismsMTT and Annexin V analysis were applied to demonstrate the influnces of different concentrations of TSA and LBH589 on the H-GMSCs.Flow assay was adopted to detect the effect of TSA and LBH589 on cell cycle.RT–PCR was performed to exam the p21 ^Waf/Cip1af/Cip1 expression of N-GMSC and H-GMSC.Treated with different concentrations of TSA and LBH589,total mRNA was extracted and underwent RT-PCR to detect p21 ^Waf/Cip1 mRNA profiles.Results:1.N-GMSC and H-GMSC were isolated from healthy gingiva and gingiva tissue from HGF,the well grown and spindle shaped cells arranged in whorled which can be observed under the microscope.Flow assay demonstrated that H-GMSCs expressed special biomarkers of mesenchymal stem cell.The results of Alizarin Red staining and Oil-O Red staining were positive.2.The MTT assay showed that TSA and LBH589 inhibited the proliferation ofH-GMSC,and the proliferation activity showed decreased with the increase concentration of HDACi.Annexin V assay showed that TSA and LBH589 promoted H-GMSC apoptosis and were dose dependent.After the treatment of H-GMSC with different concentrations of TSA and LBH589,flow cytometry showed that both TSA and LBH589 increased the G1 phase and the growth index PI was dose dependence.By semi quantitative RT-PCR,we found that the p21 ^Waf/Cip1 expression of H-GMSC was lower than that of N-GMSC.After H-GMSC was treated with TSA or LBH589,the up-regulated expression of p21^Waf/Cip1 arrested cell cycle and inhibited cell proliferation.Conclusion:1.Gingival mesenchymal stem cells（H-GMSCs）were obtained from the gingival tissue of the patients with hereditary gingival fibroma.Flow cytometry showed the expression of CD90,OCT-4,SSEA-4 and CD146 on H-GMSC surface.Alizarin red and oil red O staining were positive and H-GMSC showed multiple differentiation ability.2.Histone deacetylase inhibitors TSA and LBH589 can upregulate the p21^Waf/Cip1 in the H-GMSC,prolong the G1 phase,inhibit cell proliferation and promote cell apoptosis in a dose dependent manner.