Effects Of Chidamide And Cytosine Arabinoside On Proliferation And Apoptosis Of KG-1a Cells And Its Related Mechanisms Of PTEN/PI3K/Akt Pathway

Objective:To investigate the effects of Chidamide(CS055)and Cytosine Arabinoside(AraC)on acute myeloid leukemia cell lines KG-1a singlely and working together,and to observe their proliferation,inhibition and apoptosis.Moreover,to provide experimental basis for improving research on PTEN / PI3 K / Akt pathway.Methods:1.Thiazole Blue(MTT)Reduction Method was used to detect the cell growth inhibition after CS055,Ara-C and their combined action on the KG-1a cells in 24 hours,48 hours even 72 hours;2.Flow Cytometry(FCM)Method was used to detect the cell apoptosis in 72 hours after 1?mol/L CS055,0.1?mol/L Ara-C and their combined effect on the KG-1a cells;3.Real-time Fluorescent Quantitative Polymerase Chain Reaction(RT-PCR)was used to detect the condition of Bcl-2,PTEN and PIK-3ca-20 in 72 hours after 1?mol/L CS055,0.1?mol/L Ara-C and combined effct on the KG-1a cells.Results:1.The result of MTT showed that different concentrations of CS055 and Ara-C interfered with KG-1a cells will significantly inhibited the KG-1a.Specifically,with the increasing of the consistence and intervention,the growth inhibition became more and more obvious.Compared with the blank control group,the difference was statistically significant(P<0.01).It means CS055 and Ara-C had obvious dose dependent and time dependent when they effect on the KG-1a cells.Their IC50,CS055 and Ara-C,were 4.39?mol/L and 0.26?mol/L respectively after 72 hours.Compared with the single drug,combined effect on the inhibition were more strongly and their statistical differences were more obvious at the same time(P<0.01).2.FCM showed that after 72 hours of KG-1a cell culture,the apoptosis rate in the control group was(0.67±0.21)%,and the apoptosis rate of 1?mol/L CS055 was(16.8±0.56)%,The apoptosis rate of 0.1?mol/L L Ara-C was(9.8±0.70)%,and combined drug group was(27.9±0.91)%.Compared with the single drug,their combination group(CS055 1?mol/L + Ara-C 0.1?mol/L)increased significantly,and the difference between the groups was statistically significant(P <0.01).3.RT-PCR showed that the 1?mol/L CS055,0.1?mol/L Ara-C effected on the KG-1a cells respectively,the mRNA of BCL-2 and PIK-3ca-20 of cells decreased,and the mRNA of PTEN increased.Compared with the single drug,their combination group(CS055 1?mol/L + Ara-C 0.1?mol/L)was more significant and the statistical difference was more obvious(P <0.01).Conclusions:1.CS055 and Ara-C can inhibit the growth and promote the apoptosis of KG-1a cells,and thier combination have a synergistic effect.2.When CS055 with Ara-C interfered in the KG-1a cells together,the mRNA levels of BCL-2 and PIK-3ca-20 decreased and the mRNA levels of PTEN increased.3.CS055 combined with Ara-C may regulate PTEN / PI3 K / Akt signal pathway by activating PTEN gene expression.