[Loganetin Induces AML Cell Differentiation and Chromosomal Instability via KLHL6-Mediated CDK2 Ubiquitination Degradation].

OBJECTIVE

To investigate how loganetin, an active compound from Cornus officinalis, inhibits acute myeloid leukemia (AML) by regulating CDK2 ubiquitination-mediated degradation.

METHODS

Human AML MOLM-13 cells were treated with gradient concentrations (0, 120, 240, 480 μmol/L) of loganetin for 48 hours. CDK2 protein expression and its interaction with KLHL6 were analyzed by Western blot and co-immunoprecipitation (Co-IP), respectively. Cellular morphology was observed via Giemsa staining. Intracellular oxidative stress was evaluated by NBT assay. An AML mouse model was established by tail-vein injection of MOLM-13 cells (1×107/mL, 2×106 cells/mouse); bone marrow cells were analyzed by flow cytometry for hCD45+ and CD11b+ surface markers.

RESULTS

Loganetin dose-dependently induced KLHL6-mediated CDK2 ubiquitination and degradation. The Western blot analysis results showed that the level of CDK2 protein significantly decreased with the increase of loganetin concentration (P <0.01), Co-IP confirmed that the CDK2-KLHL6 interaction was enhanced in a dose-dependent manner. Giemsa staining revealed morphological changes (e.g., multinucleated cells, nuclear fragmentation) indicative of chromosome instability in high-concentration groups (480 μmol/L). NBT assay demonstrated elevated intracellular oxidative stress (P <0.01), accompanied by downregulation of PRDX2 and upregulation of MAFB. In vivo, treated mice showed reduced bone marrow hCD45+ AML cells (P <0.05) and increased CD11b+ differentiated cells (P <0.05), and the splenomegaly was alleviated.

CONCLUSION

Loganetin promotes AML cell differentiation by degrading CDK2 via KLHL6-mediated ubiquitination, synergistically enhancing oxidative stressand genomic instability.