Apoptosis of HL-60 human leukemia cells induced by Asiatic acid through modulation of B-cell lymphoma 2 family proteins and the mitogen-activated protein kinase signaling pathway

Wu,Qiuling; Lv,Tingting; Chen,Yan; Wen,Lu; Zhang,Junli; Jiang,Xudong; Liu,Fang

doi:10.3892/mmr.2015.3534

Apoptosis of HL-60 human leukemia cells induced by Asiatic acid through modulation of B-cell lymphoma 2 family proteins and the mitogen-activated protein kinase signaling pathway

Authors:
- Qiuling Wu
- Tingting Lv
- Yan Chen
- Lu Wen
- Junli Zhang
- Xudong Jiang
- Fang Liu
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Affiliations: Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China
Published online on: March 24, 2015 https://doi.org/10.3892/mmr.2015.3534
Pages: 1429-1434

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Abstract

The toxicities of conventional chemotherapeutic agents to normal cells restrict their dosage and clinical efficacy in acute leukemia; therefore, it is important to develop novel chemotherapeutics, including natural products, which selectively target cancer‑specific pathways. The present study aimed to explore the effect of the chemopreventive agent asiatic acid (AA) on the proliferation and apoptotic rate of the leukemia cell line HL‑60 and investigated the mechanisms underlying its anti‑tumor activity. The effect of AA on the proliferation of HL‑60 cells was evaluated using the MTT assay. Annexin V‑fluorescein isothiocyanate/propidium iodide double staining followed by flow cytometric analysis as well as Hoechst 33258 staining were used to analyze the apoptotic rate of the cells. Furthermore, changes of survivin, B‑cell lymphoma 2 (Bcl‑2), myeloid cell leukemia 1 (Mcl‑1), extracellular signal‑regulated kinase (ERK), c‑Jun N‑terminal kinase (JNK) and p38 expressions were detected by western blot analysis. AA blocked the growth of HL‑60 cells in a dose‑ and time‑dependent manner. The IC50‑value of AA on HL‑60 cells was 46.67±5.08 µmol/l for 24 h. AA induced apoptosis in a dose‑dependent manner, which was inhibited in the presence of Z‑DEVD‑FMK, a specific inhibitor of caspase. The anti‑apoptotic proteins Bcl‑2, Mcl‑1 and survivin were downregulated by AA in a dose‑dependent manner. Concurrently, AA inhibited ERK and p38 phosphorylation in a dose‑dependent manner, while JNK phosphorylation was not affected. In conclusion, the present study indicated that the p38 and ERK pathways, as well as modulation of Bcl‑2 family and survivin proteins were key regulators of apoptosis induced in HL‑60 cells in response to AA.

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