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

作　　者：Xin-xin XIONG Ju-mei LIU Xin-yao QIU Feng PAN Shang-bin YU Xiao-qian CHEN 

机构地区：[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, Tonal Medical College, Huazhong University of Science and Technology, Wuhan 430022, China

出　　处：《Acta Pharmacologica Sinica》2015年第3期362-374,共13页中国药理学报（英文版）

摘　　要：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.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.

关 键 词：piperlongumine ayurvedic medicine myeloid leukemia myelodysplastic syndrome apoptosis autophagy ROS P38 JNK N-ACETYL-L-CYSTEINE SB203580 SP600125 

分 类 号：Q785[生物学—分子生物学] O614.33[理学—无机化学]