小白菊内酯对白血病K562细胞及其干细胞的作用  

Effect of parthenolide on leukemia K562 cells and its leukemia stem cells

作　　者：易娟 陈静[1] 孙静[1] 魏虎来[1] 石建功[2] 

机构地区：[1]兰州大学基础医学院医学实验中心甘肃省新药临床前研究重点实验室,甘肃兰州730000 [2]北京协和医学院中草药物质基础与资源利用教育部重点实验室,北京100005

出　　处：《中国中药杂志》2010年第2期219-222,共4页China Journal of Chinese Materia Medica

基　　金：甘肃省自然科学研究基金项目(096RJZA034);中草药物质基础与资源利用教育部重点实验室(北京协和医学院)开放课题(0801);甘肃省新药临床前研究重点实验室(兰州大学)开放课题(GSKFKT-0802)

摘　　要：目的:研究小白菊内酯(parthenolide,PTL)对白血病K562细胞及其白血病干细胞(leukemia stem cells,LSC)的作用。方法:以白血病K562细胞为靶细胞,四氮唑蓝(MTT)比色法测定细胞增殖活性,AnnexinⅤ/PI染色法测定细胞凋亡;流式细胞术检测LSC相对含量,甲基纤维素集落形成法检测细胞的自我更新和增殖能力。结果:PTL显著抑制K562细胞的增殖,24,48,72 h的IC50分别为17.1,8.67,9.42μmol.L-1。5,10μmol.L-1PTL处理48 h,K562细胞的凋亡率分别为(49.56±5.11)%,(71.88±2.12)%。结合干细胞免疫标志分析,K562细胞中LSC样(CD34+CD38-)细胞的凋亡率分别为(52.63±4.14)%,(57.50±4.47)%。K562细胞中LSC的相对含量仅轻度增高,但高浓度(15μmol.L-1)PTL处理,LSC含量则增高15倍。0.5～4.0μmol.L-1PTL显著抑制K562细胞的集落形成能力,集落数降低24.1%～89.2%;5～15μmol.L-1PTL预处理,存活K562细胞的集落形成数增高5.0%～50.0%。结论:小白菊内酯可抑制K562细胞及其干细胞的增殖活性,并诱导其凋亡。Objective： To investigate the inhibitory and apoptosis-inducing effects of parthenolide （PTL） on human leukemia K562 cells and its leukemia stem cells（LSC）. Method： MTr assay was used to detect the proliferating activity of K562 cells, and the cellular apoptosis was assayed with Annexin V/PI double staining. Flow cytometry （ FCM ） was employed to determine the relative proportion of LSC in K562 cells. The self-renewal and proliferating potential were examined with methylcellulose colony-forming units （CFU） assay. Result： By use of MTT assay, we found PTL had significant inhibitory effect on the proliferation of K562 cells, the 50% inhibitory concentration （ IC50 ） values were 17. 1,8.67,9, 42 μmol·L^-1 for 24, 48 and 72 h, respectively. After administration with 5 μmol·L^-1 and 10 μmol·L^-1 PTL, the apoptotic rate of K562 cells was （49. 56 ＋ 5.11 ） % and （ 71.88 ＋ 2. 12 ） %, and （52. 63 ＋ 4. 14） % and （57. 50 ＋ 4. 47） % in LCS-like （ CD34^＋ CD38^- ） cells in K562 cell population, respectively. A slightly increase of relative content of LSC in K562 cells was observed. There was an 15-fold increase in the higher concentration of the PTL-treated cells. The methylcellulose colony-forming units assay showed a 24. 1% to 89. 2% decrease in the CFU of K562 cells administrated with 0. 5μmol·L^-1 to 4. 0 μmol·L^-1.PTL, and the CFU of the surviving cells increased by 5.0% to 50.0% on condition that K562 cells were pre-treated with 5 μmol·L^-1 to 15 μmol·L^-1 PTL for 48 h. Conclusion： PTL eminently inhibits proliferation of K562 cells and LSC in K562 cells, and induces the cell apoptosis.

关 键 词：白血病干细胞 小白菊内酯 集落形成 凋亡 

分 类 号：R733.7[医药卫生—肿瘤] R733.71[医药卫生—临床医学]