拓扑替康杀伤骨髓增生异常综合征原始细胞的体外和动物实验研究  

Inhibition effect of topotecan on human myelodysplastic syndrome cells in vitro and in vivo

作　　者：刘俊青[1] 章谓方[1] 金洁[1] 钱文斌[1] 

机构地区：[1]浙江大学医学院附属第一医院,浙江杭州310003

出　　处：《浙江大学学报（医学版）》2006年第2期127-131,共5页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省自然科学基金项目(M303773);浙江省科技厅社会发展重点研究项目(2004c23005)

摘　　要：目的:研究拓扑替康(TPT)在体内外对骨髓增生异常综合征(MDS)细胞株(M utz-1)细胞的杀伤作用及其机制。方法:用MTT法测定TPT对M utz-1的生长抑制作用,流式细胞仪检测细胞凋亡率,电镜观察凋亡细胞形态。对荷瘤小鼠用TPT腹腔注射,测量肿瘤大小和动物生存期作为评介TPT的抗肿瘤活性。结果:TPT显著抑制M utz-1细胞生长,呈剂量依赖性。半数生长抑制值(IC50)为272 ng/L。TPT作用48 h和72 h后细胞凋亡率分别为54.16%±4.29%和72.97%±6.12%,凋亡细胞具有典型的细胞凋亡形态学特征。用TPT治疗后5周内,荷瘤小鼠肿瘤生长速度减慢,显著低于对照组和A s2O3治疗组(P<0.05)。TPT治疗组小鼠平均生存期7周,而对照组和A s2O3组平均生存期为4周(P<0.001)。结论:在体外和动物实验中,TPT对MDS细胞株M utz-1细胞有显著的杀伤作用。Objective： To investigate the effect of topotecan （TPT） on human myelodysplastic syndrome （MDS） cells in vitro and in vivo. Methods： Cell growth was measured by a MTT assay. The percentage of cells undergoing apoptosis was determined by flow cytometry after staining with annexin V-FITC and propidium iodide. The morphology of apoptotic cells was observed by transmission electron microscopy （TEM）. Furthermore, the antitumor effect on MDS cells in xenotransplanted severe combined immunodeficiency （SCID） mice was evaluated by tumor volume and survival. Western blot was used for determining the expression of topoisomerase Ⅰ （Top1） protein. Results： The growth of Mutz-1 cells was suppressed by TPT treatment in a dose-dependent manner. The 50% inhibition in Mutz-1 cell growth （IC50） of TPT for 72 h was 272 ng/L. The percentage of apoptotic cells observed in the Mutz-1 cells after exposure to TPT （160 ng/L） in 48 h and 72 h was （54.16±4.29）% and （72.97±6.12）%, respeetively. TEM showed the characteristics of apoptosis in Mutz-1 cells treated with TPT. The xenotransplanted SCID mice treated with TPT showed inhibited tumor growth compared with control group. TPT treatment resulted in a longer survival as compared with the control group （P 〈 0.001） and with the As2O3-treated group （P 〈 0.001）. The cells exposed to TPT exhibited a time-dependent decrease of Topl protein expression. Conclusion. TPT can inhibit Mutz-1 cell growth and induce apoptosis in vitro. The downregulation of Topl may be involved in the apoptosis induced by TPT. TPT has a significant antitumor effect in vivo.

关 键 词：骨髓增生异常综合征/药物疗法 拓扑替康 拓扑异构酶Ⅰ 细胞凋亡 

分 类 号：R551.3[医药卫生—血液循环系统疾病]