Ikezoe Takayuki, Yang Yang, Bandobashi Kentaro, Saito Tsuyako, Takemoto Shigeki, Machida Hisanori, Togitani Kazuto, Koeffler H Phillip, Taguchi Hirokuni
Department of Hematology and Respiratory Medicine, KMS, Kochi Medical School, Nankoku, 783-8505 Kochi, Japan.
Mol Cancer Ther. 2005 Apr;4(4):578-86. doi: 10.1158/1535-7163.MCT-04-0277.
This study found that oridonin, a natural diterpenoid purified from Rabdosia rubescens, inhibited growth of multiple myeloma (MM; U266, RPMI8226), acute lymphoblastic T-cell leukemia (Jurkat), and adult T-cell leukemia (MT-1) cells with an effective dose that inhibited 50% of target cells (ED50) ranging from 0.75 to 2.7 microg/mL. Terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling staining showed that oridonin caused apoptosis of MT-1 cells in a time-dependent manner. We explored effects of oridonin on antiapoptotic Bcl-2 family members and found that it down-regulated levels of Mcl-1 and BCL-x(L), but not Bcl-2 protein, in both MT-1 and RPMI8226 cells. Further studies found that oridonin inhibited nuclear factor-kappa B (NF-kappa B) DNA-binding activity in these cells as measured by luciferase reporter gene, ELISA-based, and electrophoretic mobility shift assays. Oridonin also blocked tumor necrosis factor-alpha- and lipopolysaccharide-stimulated NF-kappa B activity in Jurkat cells as well as RAW264.7 murine macrophages. Of note, oridonin decreased survival of freshly isolated adult T-cell leukemia (three samples), acute lymphoblastic leukemia (one sample), chronic lymphocytic leukemia (one sample), non-Hodgkin's lymphoma (three samples), and MM (four samples) cells from patients in association with inhibition of NF-kappa B DNA-binding activity. On the other hand, oridonin did not affect survival of normal lymphoid cells from healthy volunteers. Taken together, oridonin might be useful as adjunctive therapy for individuals with lymphoid malignancies, including the lethal disease adult T-cell leukemia.
本研究发现,从冬凌草中纯化得到的天然二萜类化合物冬凌草甲素,可抑制多发性骨髓瘤(MM;U266、RPMI8226)、急性淋巴细胞性T细胞白血病(Jurkat)和成人T细胞白血病(MT-1)细胞的生长,其抑制50%靶细胞的有效剂量(ED50)为0.75至2.7μg/mL。末端脱氧核苷酸转移酶介导的dUTP缺口末端标记染色显示,冬凌草甲素以时间依赖性方式诱导MT-1细胞凋亡。我们探究了冬凌草甲素对抗凋亡Bcl-2家族成员的影响,发现它可下调MT-1和RPMI8226细胞中Mcl-1和BCL-x(L)的水平,但不影响Bcl-2蛋白水平。进一步研究发现,通过荧光素酶报告基因、基于ELISA的方法和电泳迁移率变动分析测定,冬凌草甲素可抑制这些细胞中核因子-κB(NF-κB)的DNA结合活性。冬凌草甲素还可阻断肿瘤坏死因子-α和脂多糖刺激的Jurkat细胞以及RAW264.7小鼠巨噬细胞中的NF-κB活性。值得注意的是,冬凌草甲素可降低来自患者的新鲜分离的成人T细胞白血病(3个样本)、急性淋巴细胞白血病(1个样本)、慢性淋巴细胞白血病(1个样本)、非霍奇金淋巴瘤(3个样本)和MM(4个样本)细胞的存活率,同时抑制NF-κB的DNA结合活性。另一方面,冬凌草甲素不影响健康志愿者正常淋巴细胞的存活率。综上所述,冬凌草甲素可能对包括致命疾病成人T细胞白血病在内的淋巴系统恶性肿瘤患者作为辅助治疗有用。