急性淋巴细胞白血病患儿骨髓间充质干细胞对K562和K562/AO_2细胞耐药的影响  

Effects of Bone Marrow Mesenchymal Stem Cells on Chemoresistance of K562 and K562/AO_2 Leukemia Cell Lines

作　　者：王昭霞[1,2] 邹亚伟[1] 李敏敏[1] 马玉花[1] 赵玉新[2] 陈福雄[1] 关镜明[1] 卫凤桂[1] 吴梓梁[1] 

机构地区：[1]广州医学院第一附属医院儿科,广州510120 [2]威海市文登中心医院儿科,山东威海264400

出　　处：《实用儿科临床杂志》2012年第19期1507-1510,共4页Journal of Applied Clinical Pediatrics

基　　金：广东省科技计划项目(2011B031800346)

摘　　要：目的研究急性淋巴细胞白血病(ALL)患儿骨髓间充质干细胞(MSCs)对K562和K562/AO2细胞药物耐受性的影响,并探讨其机制。方法从白血病患儿骨髓中分离、培养并鉴定MSCs;建立K562和K562/AO2细胞株与MSCs共培养体系,观察MSCs对2种细胞生长的影响;Annexin V-FITC检测一定浓度的多柔比星对2种细胞凋亡的影响;流式细胞仪测定不同培养条件下的2种细胞周期;反转录PCR分析K562和K562/AO2细胞株的黏附培养组Bcl-2和Bax基因表达;实时荧光定量PCR检测2种细胞mdr1基因转录水平。结果与单独培养的K562和K562/AO2细胞相比,与MSCs黏附共培养的白血病细胞生长较为缓慢,未见明显的对数生长期;黏附共培养的白血病细胞,G0～G1期细胞增多,S期细胞减少;K562和K562/AO2细胞株的黏附培养组Bcl-2基因表达强于悬浮组,此2组Bax基因表达均无显著差异,K562/AO2细胞Bcl-2基因相对表达要强于K562细胞株;白血病细胞单独悬浮培养组、黏附共培养组细胞mdr1基因转录水平比较差异无统计学意义(P>0.05)。结论白血病患儿MSCs能抑制白血病细胞株K562和K562/AO2生长,改变细胞周期而逃避药物的促凋亡作用,K562/AO2对多柔比星产生耐药性可能与黏附共培养后Bcl-2基因表达增强有关,而与其本身含有的mdr1基因无关。Objective To investigate the interaction between children bone marrow mesenchymal stem cells （MSCs） derived from the bone marrow of children with acute lymphoblastic leukemia（ALL） and K562 leukemia cell line, as well as the doxorubiein -tolerance K562/ AO2 cell line. Methods MSCs were harvested from bone marrow of ALL children by divided, cultured and identification, K562 or K562/AO2 cell line co - cultured to MSCs and study the effect on proliferation of leukemia cells. Every group treated with different concentrations of doxo- rubiein and measured apoptosis by AnnexinV - FITC. Measuring cell cycle after co - culture by flow eytometry. Established real - time fluores- cent quantitative polymerase chain reaction and reverse transcription polymerase chain reaction to detect muhidrug resistance protein（ mdrl ）, Bel - 2 and Bax gene. Results Unlike the culture of leukemia cells alone, co - culture with MSCs acquired lower growth rate, and without having a significant exponential phase. Increase of GO - GI phase, and decrease of S phase in both leukemia cell lines were observed by using flow eytometry. Multidrug resistance protein 1 （ MDR1 ） gene expression suggested no more significant difference between single culture and co - culture in both K562 and K562/AO2 cell lines. MSCs from child leukemia patients may restrain the growth of K562 and K562/AO2 cells, by blocking the cell cycle, and accelerating the expression of Bcl -2,while apoptosis of leukemia ceils induced by biochemieals was evaded. Otherwise ,no difference was detected in the expressions of MDR1 gene between K562 and K562/AO2 cell lines（ P 〉 0.05 ）. Conclusions ALL children MSCs suppress the growth and blocked the cell cycle of K562 and K562/AO2 cells in vitro ,and make leukemia cells to resistant eytotoxic drugs. The resistance to doxorubicin of leukemia cells may be relevant to the high expression of Bcl - 2 gene, but not mdr1 gene.

关 键 词：白血病 间充质干细胞 K562 K562/AO2 耐药 

分 类 号：R733.71[医药卫生—肿瘤]