Study In The Effect Of G9a Inhibitor BIX-01294on Cell Proliferation And Apoptosis In Molt-4Line

Objects To investigate the effect of BIX-01294on cell proliferation, apaptosis,DNA methylation and histone modulation in acute leukemia cell line Molt-4, weresearch the possible mechanism.Methods The proliferation of Molt-4cells after exposure to BIX-01294wasdetermined by MTT. Apoptotic rate was observed by flow cytometry. The change ofapoptosis related protein proCaspase-3, Bcl-2, Bax，and cell-cycle regulating factorP27、P21was observed by Western Blot. The change of histone H3acetylation, H3K9and H3K27methylation, the expression of P15and DNMT1was detected by WesternBlot.Results (1) BIX-01294inhibited Molt-4cells proliferation. Cells proliferation ratewas (86.06±4.35)%,(65.43±4.21)%,(46.23±3.17)%,(12.56±1.34)%after24hoursexposure to BIX-01294in1,2,4,8μmol/L,(p<0.05). The Cells proliferation rate was(86.06±4.35)%,(79.25±3.56)%,(70.06±3.47)%after24,48,72hours exposure toBIX-01294in1μmol/L,（p<0.05）. BIX-01294upregulated P21, P27in concentrationdependent manner.(2) BIX-01294induced apoptosis. After24hours exposure toBIX-01294in0,1,2,4μmol/L, cells apoptosis rate was (5.54±1.35)%,(15.24±2.26)%,(32.28±3.26)%,(47.52±4.37)%,(P<0.05). BIX-01294downregulated bcl-2, andupregulated Bax and proCaspase-3.(3) BIX-01294decreased histone methylatedH3K9me1, H3K9me2, H3K27me1and H3K27me2，and DNMT1, increased P15.The variation of acetylated H3and H3K9me3was not seen. Conclusions (1) BIX-01294, which is G9a inhibitor, inhibited methylation of H3K9me1, H3K9me2, H3K27me1and H3K27me2.(2) BIX-01294decreased DNAmethyltransferase DNMT1, P15was de novo.(3)BIX-01294induced cell apoptoosis,which might be consequence of downregulation of Bcl-2and uprelation of Bax andprocaspase-3.(4) BIX-01294increased the expression of P21and P27protein, andinhibited cells proliferation. BIX-01294might be a potential drug for anti-leukemia.