The Role Of Thioridazine Combined With Panobinostat In Anti-acute Myeloid Leukemia And The Underlying Mechanisms

Objective: To investigate the effects of thioridazine combined with panobinostat on the growth and proliferation of acute myeloid leukemia cells and the underlying mechanism.Methods: Acute myeloid leukemia cell lines U937 and MV-4-11 were treated with thioridazine and panobinostat at different concentrations alone or in combination for 72 h.Cell viability was detected by CCK8 assay,and cell proliferation ability was detected by soft AGAR clone formation assay.Cell apoptosis was detected by Annexin V/PI double staining flow cytometry and caspase-3 activity assay.Western blot was used to detect the protein expression and phosphorylation of necroptotic related molecules RIPK1/RIPK3/MLKL.IFN-γ was added simultaneously to detect the effects of thioridazine combined with panobinostat on the IFN-γ-induced differentiation of acute myeloid leukemia cells.Cell count assay,WrightGiemsa stain and soft AGAR clone formation test were performed to determine the combination effects on the growth,proliferation and morphology of acute myeloid leukemia cells.The expressions of SOCS1 and STAT1 were detected by western blot.Results: The results of CCK8 and soft AGAR cloning assay showed that the combination of thioridazine and panobinostat inhibited the growth and proliferation of AML cells.Flow cytometry,caspase-3 activity assay and western blot showed that the anti-leukemia effects of thioridazine combined with panobinostat were achieved by inducing apoptosis and inhibiting necroptosis mediated by RIPK1/RIPK3/MLKL.Its antileukemia effects were further enhanced by inducing the differentiation of AML cells with the addition of IFN-γ.Conclusion: Combined treatment with thioridazine enhanced the antileukemia effects of panobinostat by inducing apoptosis and inhibiting necroptosis,and promoted the differentiation of AML cells induced by IFN-γ.Figures 7,tables 9,references 63...