Molecular mechanism of the G1 arrest and cellular senescence induced by LEE011, novel CDK4/CDK6 inhibitor, in acute myeloid leukemia cells

ABSTRACT: Background CDK4 and CDK6 are frequently over expressed or hyper activated in human cancers. Due to the importance of CDK4/6 activity in cancer cells, CDK4/6 inhibitors have emerged as promising candidates for cancer treatment. LEE011, a novel inhibitor of CDK4/6, until now the molecular function of LEE011 in acute myeloid leukemia is still unknown. Methods AML cell growth was assessed by CCK-8 and annexin V/propidium iodide staining followed by flow cytometry detected apoptosis in cell culture. AML cell senescence was assessed by β-Galactosidase Staining, p16INK4a expression and HP1α Staining analysis. Gene expressions of LEE011-treated tumor cells were analyzed using an Arraystar Human LncRNA Array. The Multi Experiment View cluster software analyzed the expression data. Differentially expressed genes from the cluster analyses were imported into the GO and KEGG pathway analysis. Results Our studies demonstrated that LEE011 inhibited proliferation of leukemia cells and do not induce apoptosis. Hoechst 33342 staining analysis showed that DNA fragmentation and abnormal nuclear cells was observed after LEE011 treatment. Cell cycle analysis showed in these eight acute leukemia cells, LEE011 induce the cell cycle G1 arrest very significantly, except THP-1 cells. Cell senescence β-Galactosidase Staining analysis, p16INK4a expression and HP1α Staining analysis all showed that LEE011 treatment can induce the cell senescence of AML cells. LncRNA microarray was used to identify 2083 differently expressed mRNAs and 3224 differentially expressed lncRNAs in LEE011-treated HL-60 cells compared with control group. Molecular function analysis showed that LEE011 induced cellular senescence in leukemia cells partially through down regulation the transcriptional expression of MYBL2. Conclusions In this study, we showed for the first time that LEE011 treatment resulted in inhibition of cell proliferation and induction of G1 arrest and cellular senescence in AML cells. LncRNA microarray was used to identify differentially expressed genes and lncRNAs in LEE011-treated HL-60 cells and we demonstrated that LEE011 induce cellular senescence partially through down regulation the expression of MYBL2. These results may provide new insights into the molecular mechanism of LEE011-induced cellular senescence; however, further research will be required to determine the underlying details. Taken together, our findings firstly suggest that LEE011 may act as new candidate’s drug for AML.

ORGANISM(S): Homo sapiens

PROVIDER: GSE81060 | GEO | 2017/06/13

SECONDARY ACCESSION(S): PRJNA320395

REPOSITORIES: GEO