Study on the mechanism of diterpenoid DP from Euphorbia fischeriana against leukemia
- VernacularTitle:狼毒大戟中二萜类化合物DP抗白血病的作用机制研究
- Author:
Liwei MA
1
,
2
;
Zhe CHEN
3
;
Wenbao WANG
4
;
Jinling ZHANG
1
;
Hongtao ZHANG
5
;
Pengling GE
2
;
Jicheng LIU
1
Author Information
1. Research Institute of Medicine and Pharmacy （Postdoctoral Workstation），Qiqihar Medical University，Heilongjiang Qiqihar 161006，China
2. Mobile Postdoctoral Station，Heilongjiang University of Chinese Medicine，Harbin 150040，China
3. School of Public Health，Qiqihar Medical University，Heilongjiang Qiqihar 161006，China
4. School of Pharmacy，Qiqihar Medical University，Heilongjiang Qiqihar 161006，China
5. Dept. of Obstetrics and Gynecology，the Third Affiliated Hospital of Qiqihar Medical University，Heilongjiang Qiqihar 161099，China
- Publication Type:Journal Article
- Keywords:
Euphorbia fischeriana;
12-deoxyphorbol-13-palmitate;
human myeloid leukemia HL60 cells;
phosphoinositide 3-
- From:
China Pharmacy
2023;34(7):825-831
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To explore whether diterpenoid 12-deoxyphorbol-13-palmitate （DP） from Euphorbia fischeriana can exert anti-leukemia effects through the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signal pathway， and to provide experimental evidence for developing it into a new anti-leukemia drug. METHODS Using LY294002 （PI3K specific inhibitor） as tool drug， the effects of 24 h DP treatment on the proliferation and apoptosis of human myeloid leukemia HL60 cells were detected by MTT method， Annexin Ⅴ-FITC/PI staining and AO-EB staining. ELISA method was used to detect lactic dehydrogenase （LDH） release and the activities of cysteinyl aspartate specific proteinase 3 （caspase-3） and caspase-9. The transcriptional level of caspase-3， caspase-9， forkhead box O3a （FoxO3a） and B cell lymphoma 2 interacting mediator of cell death （Bim） mRNA were detected by real-time quantitative polymerase chain reaction （qRT-PCR）. The protein expression of phosphorylated FoxO3a （p- FoxO3a） and phosphorylated Akt （p-Akt） were detected by Western blot method. The nuclear translocation of FoxO3a protein was detected by immunostaining combined with laser confocal microscopy. RESULTS 10 μmol/L DP and 10 μmol/L DP+LY294002 could inhibit the proliferation and induce the apoptosis of HL60 cells （P＜0.01）. After treatment of 5， 10， 20 μmol/L DP， HL60 cells showed typical morphological characteristics of apoptosis； DP could significantly increase the levels of LDH release and the activities of caspase-3 and caspase-9 （P＜0.05 or P＜0.01）， in dose-dependent manner. After treatment of 10 μmol/L DP and 10 μmol/L DP+LY294002， the transcriptional levels of caspase-3， caspase-9 and Bim mRNA were increased significantly （P＜0.05 or P＜0.01）， and transcriptional level of FoxO3a mRNA and protein expressions of p-FoxO3a and p-Akt were decreased significantly （P＜0.05 or P＜0.01）. Nuclear translocation changes were observed in FoxO3a protein in 10 μmol/L DP+LY294002 group， and the change was more significant than that of LY294002 group. CONCLUSIONS DP can inhibit the proliferation and induce the apoptosis of HL60 cells via inhibiting PI3K/Akt signaling pathway.
