K562细胞株侧群细胞分离及耐药蛋白和细胞周期的表达

背景：血液肿瘤细胞中的侧群细胞能够逃避细胞周期化疗药物的作用,可能与白血病复发有关。目的：鉴定人慢性粒细胞白血病细胞株K562中是否存在侧群细胞,并观察侧群细胞亚群与非侧群细胞亚群耐药蛋白、细胞周期表达的差异。方法：采用流式细胞术检测K562细胞株中是否存在侧群细胞;并分析K562侧群细胞和非侧群细胞两亚群间耐药蛋白P-gp、ABCG2以及细胞周期的表达情况。结果与结论：K562细胞株中均存在侧群细胞,这群细胞比例均少。侧群细胞亚群耐药蛋白ABCG2表达高于非侧群细胞亚群（P〈0.05）,两亚群耐药蛋白P-gp中表达差异无显著性意义;侧群细胞亚群中G0/G1期细胞占80%,非侧群细胞亚群中G0/G1期细胞占43.7%。证实K562细胞株中确实存在较少比例的侧群细胞,其和主群细胞在耐药蛋白表达上有异质性,大部分处于静止期,可能富含和肿瘤耐药有关的白血病干细胞。

Isolation of side population cells in leukemia K562 cell lines and the expression of resistance protein and cell cycle in two subpopulations

BACKGROUND：It is possible that leukemia relapse is related with side population（SP） cells,which can escape cell cyles phase specific chemotherapeutic drugs.OBJECTIVE：To isolate and preliminarily identify whether the human chronic myeloid leukemia cell line-K562 contains SP cells or not,and to further research K562 SP cells and study the expression of multidrug resistance proteins and DNA content in two subpopulations.METHODS：Flow cytometry with UV excitation light was used to detect the percentage of SP cells in logarithmic growth period K562,which were then sorted by the fluorescence activating cell sorter,and then SP and non-SP subpopulations were collected.The expression of multidrug resistance proteins were examined by flow cytometry technique in two subpopulations.DNA content of two subpopulations was examined by flow cytometry.RESULTS AND CONCLUSION：The K562 cell line contained SP cells,and the proportion of SP cells was much lower.Although there were no differences between these two subpopulations in P-gp expression,ABCG2＋ cells expression in the SP subpopulation was significant higher than that in the non-SP subpopulation（P 0.05）.The G0/G1 phase cells in the K562 SP subpopulation accounted for about 80% of the total cells.According to the significant differences in the expression of MDR proteins between the SP subpopulation and the non-SP subpopulation,and the majority in a quiescent period,it is possible that leukemia stem cells are enriched in SP cells.