Study Of The Effects Of Chidamide On Proliferation?Apoptosis And Related Mechanism Of Hedgehog Signaling In K562/G01 Cells

Objectives:The purpose of this study was to observe the effect of chidamide on the proliferation and apoptosis in K562/G01 cell which is Imatinib-resistaIlt chronic myeloid leukemia cell lines and detecting the expression of related genes in Hedgehog pathway.We discuss the possible mechanism of chidamide on K562/G01 cells for the purpose of providing the new ways and the theory basis for chidamide's application in clinical treatment of Imatinib-resistaIlt chronic myeloid leukemia.Methods:The K562/G01 cells were respectively treated by chidamide with different concentrations in 0-60?mol/L for 24 h,48h and 72 h.The cells proliferation inhibition rates was measured by methyl thiazolyl tetrazolium(MTT),and SPSS 20.0 software was used to calculate the half inhibitory concentration value(IC50)of chidamide on K562/G01 cells.The apoptosis rate was detected by flow cytometry.The m RNA expressions of Gli1?BCR/ABL and CyclinD2 were analyzed by real-time fluorescent quantitative polymerase chain reaction(RQ-PCR).The protein levels of BCR/ABL?Gli1?CyclinD2 and Acety-histone H3 were determined by Western Blot.Experiments were all repeated 3 times,The results were expressed with mean±SEM(x ±s,%),and P<0.05 was considered statistically significant.Results:1.The proliferation of K562/G01 cells were significantly inhibited by chidamide and the inhibition rates was dose and time dependent.When the drug concentration was increased from 2.5?mol/l to 60?mol/l,the inhibition rate of cell proliferation was increased from(10.57±0.48)% to(63.08±0.79)% at 24 hours,and the inhibition rate was increased from(27.10±0.49)% at 48 hours.(74.67±1.78)%;72-hour proliferationinhibition rate was increased from(32.61±1.44)% to(84.48±3.04)%.2.After K562/G01 cells respectively treated with 0,10,30?mol/L chidamide for48 hours and 72 hours,the corresponding apoptosis rates detected by Annexin V/PI flow cytometry and whose at 48 hours were(1.1±0.36)%?(13.8±1.50)%?(31.2±2.79)% and at 72 h hours were(4.4±1.45)% ?(33.7±1.72)% ?(62.4±2.68)%,the apoptosis rates of chidamide was dose dependent.3.After K562/G01 cells respectively treated with chidamide,the dates detected by RQ-PCR showed that m RNA expression levels of Gli1 ? CyclinD2 ? BCR/ABL decreased obviously.4.After K562/G01 cells treated with chidamide,the dates detected by Western Blot showed that the expression level of BCR/ABL?Gli1?CyclinD2 protein decreased obviously and the level of acetylated instone H3 was significantly increased.Conclusions:1.The inhibition of proliferation and apoptosis of K562/G01 cells by chidamide in a concentration and time dependent manners.2.The inhibition of proliferation and apoptosis of K562/G01 cells by chidamide is related to BCR/ABL gene inhibition and histone H3 acetylation.3.The inhibition of proliferation and apoptosis of K562/G01 cells by chidamide is regulating the activity of Hedgehog signaling pathway.