Mechanism of evodiamine inducing cell cycle arrest and apoptosis in human erythroleukemia K562 cells through inhibiting histone deacetylase 6 / 中草药

ABSTRACT

Objective: To explore the mechanism of evodiamine (Evo) inducing cell cycle arrest and apoptosis in K562 cells. Methods: The effect of Evo on proliferation of K562 cells was measured by Cell Counting Kit-8 assay (CCK-8 assay), and cell cycle distribution and apoptosis were determined by flow cytometry (FCM). Chemical colorimetry assay was used to examine the activity of histone modification enzymes. The expression levels of histone deacetylase 6 (HDAC6), Cyclin D1, CDK4, Bcl-2, Bax, Cleaved Caspase-3, ERK, p-ERK, p38, and p-p38 proteins were ascertained by Western blotting. Results: The proliferation of K562 cells was inhibited by Evo (1-16 μmol/L) in a dose- and time-dependent manner. FCM analyses revealed that Evo induced cell-cycle arrest in G0/G1 phase in K562 cells. The apoptosis rates of K562 cells were (11.47 ± 1.05)%, (12.77 ± 0.79)%, and (18.58 ± 1.37)% respectively after induced by Evo with different concentration (2, 4, and 8 μmol/L), which showed statistically significant difference compared with the control group (2.79 ± 1.01)% (P < 0.01). The activity of HDACs was reduced after treated with Evo (2, 4, and 8 μmol/L). Western blotting assay showed that the expression of Bax, Cleaved Caspase-3, p38, and p-p38 proteins increased, while CDK4, Cyclin D1, Bcl-2, HDAC6, ERK, and p-ERK proteins down-reguation after induced by Evo. Conclusion: Evo can induce cell cycle arrest and apoptosis in K562 cells through the inhibition of HDAC6.