Piperlongumine induces apoptotic and autophagic death of the primary myeloid leukemia cells from patients via activation of ROS-p38/JNK pathways

• Published in: Acta pharmacologica Sinica 2015-03, Vol.36 (3), p.362-374
• Main Authors: Xiong, Xin-xin, Liu, Ju-mei, Qiu, Xin-yao, Pan, Feng, Yu, Shang-bin, Chen, Xiao-qian
• Format: Article
• Language: English
• Subjects: Antineoplastic Agents, Phytogenic - pharmacology; Antioxidants - pharmacology; Apoptosis - drug effects; Apoptosis Regulatory Proteins - metabolism; Autophagy - drug effects; Biomedical and Life Sciences; Biomedicine; Cell Survival - drug effects; Dioxolanes - pharmacology; Dose-Response Relationship, Drug; Enzyme Activation; Humans; Immunology; Inhibitory Concentration 50; Internal Medicine; JNK; JNK Mitogen-Activated Protein Kinases - antagonists & inhibitors; JNK Mitogen-Activated Protein Kinases - metabolism; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - drug therapy; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - enzymology; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - pathology; Leukemia, Myeloid, Acute - drug therapy; Leukemia, Myeloid, Acute - enzymology; Leukemia, Myeloid, Acute - pathology; Medical Microbiology; Original; original-article; Oxidative Stress - drug effects; p38; p38 Mitogen-Activated Protein Kinases - antagonists & inhibitors; p38 Mitogen-Activated Protein Kinases - metabolism; Pharmacology/Toxicology; Phosphorylation; Primary Cell Culture; Protein Kinase Inhibitors - pharmacology; Reactive Oxygen Species - metabolism; Signal Transduction - drug effects; Tumor Cells, Cultured; Vaccine; 凋亡诱导; 患者; 抗氧化剂; 白血病细胞; 细胞凋亡; 自噬基因
• ISSN: 1671-4083; 1745-7254
• DOI: 10.1038/aps.2014.141

Aim： To investigate the effects of piperlongumine （PL）, an anticancer alkaloid from long pepper plants, on the primary myeloid leukemia cells from patients and the mechanisms of action. Methods： Human BM samples were obtained from 9 patients with acute or chronic myeloid leukemias and 2 patients with myelodysplastic syndrome （MDS）. Bone marrow mononuclear cells （BMMNCs） were isolated and cultured. Cell viability was determined using MTT assay, and apoptosis was examined with PI staining or flow cytometry. ROS levels in the cells were determined using DCFH-DA staining and flow cytometry. Expression of apoptotic and autophagic signaling proteins was analyzed using Western blotting. Results： PL inhibited the viability of BMMNCs from the patients with myeloid leukemias （with IC50 less than 20 μmol/L）, but not that of BMMNCs from a patient with MDS. Furthermore, PL （10 and 20 μmol/L） induced apoptosis of BMMNCs from the patients with myeloid leukemias in a dose-dependent manner. PL markedly increased ROS levels in BMMNCs from the patients with myeloid leukemias, whereas pretreatment with the antioxidant N-acetyl-L-cysteine abolished PL-induced ROS accumulation and effectively reduced PL-induced cytotoxicity. Moreover, PL markedly increased the expression of the apoptotic proteins （Bax, Bci-2, and caspase-3） and autophagic proteins （Beclin-1 and LC3B）, and phosphorylation of p38 and JNK in BMMNCs from the patients with myeloid leukemias, whereas pretreatment with the specific p38 inhibitor SB203580 or the specific JNK inhibitor SP600125 partially reversed PL-induced ROS production, apoptotic/autophagic signaling activation and cytotoxicity. Conclusion： Piperlongumine induces apoptotic and autophagic death of the primary myeloid leukemia cells from patients via activation of ROS-p38/JNK pathways.