Zhang Xia, Zhao Zichu, Yi Sha, Wen Lu, He Jing, Hu Jingyu, Ruan Jun, Fang Jun, Chen Yan
Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, People's Republic of China.
Anticancer Drugs. 2017 Aug;28(7):723-738. doi: 10.1097/CAD.0000000000000494.
Nucleophosmin (NPM1), a restricted nucleolar localization protein, shuttles between the nucleus and the cytoplasm. Mutated (Mt)-NPM1 protein, which has aberrant cytoplasmic dislocation of nucleophosmin, occurs in approximately one-third of acute myeloid leukemia cases. Deguelin, a rotenoid isolated from several plant species, is a strong antitumor agent. NOD/SCID mice xenografted with human Mt-NPM1 OCI/AML3 cell lines served as in-vivo models. Wright-Giemsa staining and flow cytometry analysis were used for differentiation assays. Associated molecular events were assessed by western blot and histological analyses. Kaplan-Meier estimates were used to calculate survival. Deguelin toxicity in mice was assessed by immunohistochemistry staining and serum markers. Clinical samples were differentiated by flow cytometry analysis. Deguelin induced differentiation by downregulating the Mt-NPM1 protein levels, which was accompanied by a decrease in SIRT1, p21, and HDAC1 and an increase in CEBPβ and granulocyte colony-stimulating factor receptor protein expression levels. A low-deguelin dose prolonged survival compared with the control group, and there were no apparent lesions to the brain, liver, heart, and kidney in vivo. In clinical samples, deguelin induced the differentiation of fresh blasts with Mt-NPM1 protein, but not with the wild-type NPM1 protein. Taken together, these findings further provide new evidence that the Mt-NPM1 protein plays an important role in inducing differentiation in vivo and in vitro. Mutated NPM1 protein may be a therapeutic target of deguelin in acute myeloid leukemia with the NPM1 mutation.
核磷蛋白（NPM1）是一种局限于核仁定位的蛋白质，在细胞核和细胞质之间穿梭。核磷蛋白发生突变（Mt）的蛋白质在急性髓系白血病病例中约占三分之一，其核磷蛋白在细胞质中出现异常移位。鱼藤素是从几种植物中分离出的一种鱼藤酮类化合物，是一种强效抗肿瘤剂。用人Mt-NPM1 OCI/AML3细胞系异种移植的NOD/SCID小鼠作为体内模型。采用瑞氏-吉姆萨染色和流式细胞术分析进行分化检测。通过蛋白质印迹和组织学分析评估相关分子事件。采用Kaplan-Meier估计法计算生存率。通过免疫组织化学染色和血清标志物评估鱼藤素对小鼠的毒性。通过流式细胞术分析对临床样本进行鉴别。鱼藤素通过下调Mt-NPM1蛋白水平诱导分化，同时伴随着SIRT1、p21和HDAC1的减少以及CEBPβ和粒细胞集落刺激因子受体蛋白表达水平的增加。与对照组相比，低剂量鱼藤素可延长生存期，且在体内对脑肝肾无明显损伤。在临床样本中，鱼藤素可诱导带有Mt-NPM1蛋白的新鲜原始细胞分化，但不能诱导带有野生型NPM1蛋白的原始细胞分化。综上所述，这些发现进一步提供了新的证据，表明Mt-NPM1蛋白在体内和体外诱导分化中起重要作用。突变的NPM1蛋白可能是鱼藤素治疗NPM1突变型急性髓系白血病的靶点。
