5-FU富集人白血病细胞系KG-1a中肿瘤干细胞样亚群细胞  

Enrichment of Tumor Stem-like Subpopulation Cells with 5-FU in Human Leukemia Cell Lines KG-1a

作　　者：杨松[1] 钟晓明[1] 张伶[1] 高玉洁[1] 骆展鹏[1] 何鹏[1] 

机构地区：[1]重庆医科大学医学检验系临床检验诊断学省部共建教育部重点实验室,重庆市重点实验室,重庆400016

出　　处：《细胞生物学杂志》2008年第6期786-790,共5页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(No.30872418);重庆市教委科学技术研究基金(No.KJ050309)资助项目~~

摘　　要：探讨利用细胞周期特异性药物5-氟尿嘧啶(5-fluorouracil,5-FU)从人白血病细胞系KG-1a中富集肿瘤干细胞样亚群细胞。体外药物敏感试验确定5-FU的最佳作用浓度和作用时间;KG-1a细胞经5-FU药物处理后,流式细胞术检测存活细胞群中CD34+CD38-亚群细胞比例;吖啶橙染色观察细胞内的核酸组成;RT-PCR半定量检测细胞内三磷酸腺苷结合转运蛋白G超家族成员2(ATP-binding cassette superfamily Gmember2,ABCG2)mRNA的表达;半固体培养观察细胞的集落形成能力。结果显示,50μg/ml5-FU作用KG-1a细胞4天后,CD34+CD38-亚群细胞比例提高10倍以上;吖啶橙染色可见大部分细胞核酸以发出绿色荧光的DNA为主,RNA含量低;此类细胞高表达ABCG2 mRNA水平;而药物处理后细胞集落形成数量较未处理细胞明显减少。研究结果表明,利用5-FU能够杀死增殖期细胞的性质,成功建立体外药物筛选富集人白血病细胞系KG-1a中肿瘤干细胞样亚群细胞的方法。To enrich tumor stem cell-like subpopulation in human leukemia cell lines KG-la with 5- fluorouracil （5-FU）, optimal concentration and effect time of 5-FU were determined with in vitro drug sensitivity assay. The CD34^＋CD38^- subpopulation in the enriched subpopulation of KG-1a cells was detected with flow cytometry. The RNA content of enriched subpopulation cells was measured by using acridine orange staining, and their expression of ABCG2 mRNA by RT-PCR. The colony-forming ability of this subpopulation cells was evaluated by semi-solid culturing. It was indicated through our research that after treatment of KG-1a cells with 50 μg/ml 5- FU for 4 days, the proportion of CD34^＋CD38^- cells in enriched subpopulation was elevated by more than 10 folds; The RNA content of enriched subpopulation cells was low, but the ABCG2 mRNA was highly expressed. The colony forming ability of enriched subpopulation cells was lower than of non-enriched KG-1a cells. Our study suggested that utilizing 5-FU for killing most proliferating cells, we successfully established the method in which selecting and enriching tumor stem-like subpopulation cells in human leukemia cell lines KG-1a with 5-FU.

关 键 词：白血病干细胞 5-氟尿嘧啶 KG-1a细胞系 集落形成 耐药 

分 类 号：R733.7[医药卫生—肿瘤]