Antitumor Effects And Molecular Mechanisms Of Novel HDAC Inhibitors Fa In T-lymphoma Cell Lines Jurkat

Objective:To examine antitumor effects of novel HDAC inhibitors Fa and explore anti-proliferative molecular mechanisms in human T-lymphoma cell lines Jurkat.Method:1?Using CCK-8 assays to detect different concentration of Fa to Jurkat cell lines proliferation: the concentration of Fa,respectively,for 1.56?3.125?6.25?12.5?25 and50 ?M.Observe Jurkat cells inhibition rate and calculate half growth inhibition concentration?IC50?,when cells were stimulated for 24?48 and 72 h respectively;2?Examine the effects on the cell cycle arrest by flow cytometry,when the cells were stimulated for 72h;Observe the change of cell cycle,when Jurkat cells were stimulated for 24?48 and 72 h respectively by 25?M Fa;3?Using western blotting to detect the protein expression of cyclin D ? CDK4 and p21^cip/WAF;4 ? The genes expression of HDAC1?HDAC2 and HDAC3 were detected by RT-PCR method;5?The software Prism 5.0 was used to analyze the data.Mean±standard deviation expresses grouped data.Two groups t-test and one-way analysis of variance were used for comparing means.P<0.05 was indicated statistical significance.Results:CCK-8 test results showed that in normal cells as constract,half growth inhibition concentration?IC50?respectively were 88.72 ± 0.13?25.45 ± 0.03?12.21 ± 0.07 ?M,after the Jurkat cells were stimulated for 24?48 and 72 h by different concentrations of Fa.When the cells were stimulated for 72 h,the percentage of cell viability decreased from 95.6±2.57% to 11.4±1.41%,which was time and dose-dependent;Flow cytometry results showed that,different concentrations of Fa acting on Jurkat cells for72 h cells can produce G0/G1 cell cycle arrest,which was dose-dependentand.On the concentration of 25?M Fa,with the prolongation of time,the percentage of cells in G0/G1 phase increased from 42.54 ± 2.11% to 61.42 ± 0.59%.Protein expression of p21^cip/WAF increased,and protein expression of cyclin D,CDK4 was down-regulated;Fa drugs can effectively inhibit the activity of HDAC1,HDAC2,HDAC3.Conclusion:Fa has a certain anti-tumor activity on T cell lymphoma cell lines,and its mechanism is related to the induction of cell cycle arrest and up regulation of tumor suppressor gene p21^cip/WAF expression.