Study On Inhibitory Efficacy Of The Histone Acetylation Enzyme To The Cells Of Glioma U251

Glioma as one of the most challenging malignant tumors in centralnervous system,accounts for40%of the intracranial tumours.Although thereare multiple treating strategies to glioma at present,each one has itsshortcomings.The invasiveness of glioma cells determines it’s difficult todistinguish the bundary of glioma tissue from the normal cerebral tissueintraoperatively,radiotherapy is harmful to the normal cerebral tissue and thedrugs in common use are not so effective because of blood–brain-barrier anddrug resistance.So,finding a promising chemotherapeutic agent is urgent at themoment.We find that chemotherapeutics targeting histone acetyltransferase（HAT）is potent in glioma inhibiting.The differentiation and metabolism of a normal cell depends on theequlibrium of histone HAT and histone deacetylase（HDAC）. A cell with theinterfering of the equilibrium of HAT and HDAC will prone to cancerization.HDAC inhibitors,by the way of hindering cell cycle,inducing celldifferentiation and apoptosis and inhibiting tumor angiogenesis could impedingglioma.,But the mechanism is unclear.In this study we aimed to investigate the effect of HDAC inhibitors toglioma cells and the related genes and explore the mechanism preliminarily.Purposes: By observing the impact of HDAC inhibitor LBH589on theproliferation rate of human glioma cells U251in vitro,we try to explore a newdrug in the clinical management of glioma and its theoretical basis.Methods: Cultivate the U251glioma cells in vitro,and select the cells inthe logarithmic growth period.The cells were divided into the Controltrol group,LBH5890.5μmol.L^-1Group, LBH5891μmol.L^-1Group and LBH5892μmol.L^-1Group.Detect the cell viability of each group by MTT assay, the cell apoptosis of U251cells by the way of Hoechst33342, the level ofmitochondrial apoptosis relating protein Bcl-2/Bax in U251cells and theexpression of apoptosis relating protein Caspase-3by the method of Westernblot, the mRNA of Bax/Bcl-2and caspase-3through RT-PCR.Results:1、MTT showed Compared with the Control group, the proliferation ratedcreases obviously in LBH5891μmol.L^-1Group.2、Compared with the Control group, in LBH5890.5μmol.L^-1Group,LBH5891μmol.L^-1Group and LBH5892μmol.L^-1Group, the Bcl-2/Bax level ofthe U251cells increased orderly （P<0.05）, and the expression of Caspase-3increased（P<0.05） by the method of RT-PCR,which showed that themitochondrial pathway participates in the cell apoptosis.3、Hoechst33342showed Compared with the Control group, in LBH589when the drug Controlcentration reachs2umol.L^-1,cells apoptosisedprofoundly.4、RT-PCR showed compared with the Control group,the mRNA ofBax/Bcl-2and caspase-3increases obviously.Conclusions:1、HDAC inhibitor LBH589can inhibit the proliferation and growth ofglioma cell U251.2、HDAC inhibitor LBH589may induce the apoptosis of glioma cellU251through the mitochondrial pathway.