Antitumor effect of osthole on human B-cell acute lymphoblastic leukemia 697 cells and its mechanism
10.3781/j.issn.1000-7431.2019.11.439
- Author:
Cong ZHU
1
Author Information
1. Department of Pediatrics, Affiliated Hospital of Binzhou Medical University
- Publication Type:Journal Article
- Keywords:
Apoptosis;
Autophagy;
Cell proliferation;
Experimental;
Leukemia;
Osthole
- From:
Tumor
2019;39(2):91-98
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the antitumor effect of osthole on human B-cell acute lymphoblastic leukemia (B-ALL) 697 cells and its possible mechanism. Methods: After B-ALL 697 cells were treated with different concentrations (8, 16, 32, 64 and 128 μmol/L) of osthole, the inhibition rate of cell proliferation was detected by CCK-8 assay. After B-ALL 697 cells were treated with 8 and 32 μmol/L osthole, the apoptosis was detected by FCM, the expressions of apoptosis-associated molecule Bcl-2 and Bax were detected by real-time fluorescent quantitative PCR and Western blotting. After B-ALL 697 cells were treated with 8 and 32 μmol/L osthole alone or in combination with autophagy inhibitor 3-methyladenine (3-MA), the intracellular mean fluorescent intensity (MFI) was detected by FCM [stained by monodansylcadaverine (MDC)] to reflect the autophagy. The expressions of autophagy-associated molecule Beclin 1 mRNA and protein was detected by real-time fluorescent quantitative PCR and Western blotting, respectively. Results: The proliferation of B-ALL 697 cells in 8, 16, 32, 64 or 128 μmol/L osthole treatment group was significantly inhibited in a dose-and time-dependent manner (all P < 0.05). The apoptosis rate of B-ALL 697 cells in 8 or 32 μmol/L osthole treatment group was significantly increased (both P < 0.01), the expression levels of Bax mRNA and protein were increased (all P < 0.05), while the expression levels of Bcl-2 mRNA and protein were decreased (all P < 0.05). The MDC MFI of B-ALL 697 cells in 8 or 32 μmol/L osthole treatment group was increased (both P < 0.01), and the expression levels of Beclin 1 mRNA and protein were significantly increased (all P < 0.05). There was no significant change in MDC MFI and the expressions of Beclin 1 mRNA and protein after treatment with 3-MA in combination with osthole (all P > 0.05). Conclusion: Osthole inhibits the proliferation, and induces the apoptosis and autophagy of B-ALL 697 cells. The mechanism of promoting apoptosis may be related to the up-regulation of Bax expression and the down-regulation of Bcl-2 expression. Beclin 1 participates in the autophagy.
