title: Piperlongumine induces apoptotic and autophagic death of the primary myeloid leukemia cells from patients via activation of ROS-p38/JNK pathways.
abstract: OBJECTIVE
  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, Bcl-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.
  
  
  CONCLUSIONS
  Piperlongumine induces apoptotic and autophagic death of the primary myeloid leukemia cells from patients via activation of ROS-p38/JNK pathways.
authors.affiliation: Department of Pathophysiology, School of Basic Medicine; Key Laboratory of Neurological Diseases, Ministry of Education; Institute of Brain Research, Huazhong University of Science and Technology, Wuhan 430030, China.
authors.forename: Xin-xin
authors.initials: XX
authors.lastname: Xiong
authors.affiliation: Department of Pathophysiology, School of Basic Medicine; Key Laboratory of Neurological Diseases, Ministry of Education; Institute of Brain Research, Huazhong University of Science and Technology, Wuhan 430030, China.
authors.forename: Ju-mei
authors.initials: JM
authors.lastname: Liu
authors.affiliation: Department of Pathophysiology, School of Basic Medicine; Key Laboratory of Neurological Diseases, Ministry of Education; Institute of Brain Research, Huazhong University of Science and Technology, Wuhan 430030, China.
authors.forename: Xin-yao
authors.initials: XY
authors.lastname: Qiu
authors.affiliation: 1] Department of Pathophysiology, School of Basic Medicine; Key Laboratory of Neurological Diseases, Ministry of Education; Institute of Brain Research, Huazhong University of Science and Technology, Wuhan 430030, China [2] Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Pan
authors.affiliation: Department of Pathophysiology, School of Basic Medicine; Key Laboratory of Neurological Diseases, Ministry of Education; Institute of Brain Research, Huazhong University of Science and Technology, Wuhan 430030, China.
authors.forename: Shang-bin
authors.initials: SB
authors.lastname: Yu
authors.affiliation: Department of Pathophysiology, School of Basic Medicine; Key Laboratory of Neurological Diseases, Ministry of Education; Institute of Brain Research, Huazhong University of Science and Technology, Wuhan 430030, China.
authors.forename: Xiao-qian
authors.initials: XQ
authors.lastname: Chen
mesh_terms: D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D001343:Autophagy / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D004148:Dioxolanes / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D006801:Humans; D020128:Inhibitory Concentration 50; D048031:JNK Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D018384:Oxidative Stress / Q000187:drug effects*; D010766:Phosphorylation; D061251:Primary Cell Culture; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010881:Piperidones
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D000975:Antioxidants; D051017:Apoptosis Regulatory Proteins; D004148:Dioxolanes; D048031:JNK Mitogen-Activated Protein Kinases; D047428:Protein Kinase Inhibitors; D017382:Reactive Oxygen Species; D048051:p38 Mitogen-Activated Protein Kinases; C498077:piperlongumine; D010881:Piperidones