MicroRNA-223在淋巴细胞白血病原代细胞中表达及作用机制的研究  

Expression of MicroRNA-223 in Lymphocytic Leukemia Cells and Its Action Mechanism

作　　者：南祯[1] 梁勇[1] 付蓉[1] 刘惠[1] 阮二宝[1] 王晓明[1] 王国锦[1] 瞿文[1] 刘鸿[1] 吴玉红[1] 宋嘉[1] 邢莉民[1] 关晶[1] 李丽娟[1] 王化泉[1] 邵宗鸿[1] 

机构地区：[1]天津医科大学总医院血液科,天津300052

出　　处：《中国实验血液学杂志》2013年第3期556-561,共6页Journal of Experimental Hematology

摘　　要：本研究旨在探讨急性淋巴细胞白血病(ALL)、慢性淋巴细胞白血病(CLL)原代细胞中MicroRNA-223与LMO2基因的表达水平及MicroRNA-223的作用机制。应用人淋巴细胞分离液分选ALL、CLL患者及正常人骨髓中淋巴细胞,在ALL、CLL患者骨髓淋巴细胞中转染MicroRNA-223的类似物靶向升高细胞中MicroRNA-223的表达,在正常人骨髓淋巴细胞中转染MicroRNA-223抑制物靶向敲低细胞中MicroRNA-223的表达。转染后培养72 h,用RT-PCR方法检测转染前后MicroRNA-223和LMO2表达量及其相关性,并用流式细胞术进一步观察细胞凋亡和细胞周期的变化。结果表明,转染MicroRNA-223类似物前,ALL、CLL患者中MicroRNA-223表达水平为(433.11±144.88),LMO2水平为(807.10±238.41),正常人转染MicroRNA-223抑制物前,MicroRNA-223表达水平为(949.59±267.39),LMO2的表达为(455.32±176.83);MicrRNA-223的表达在正常人中明显高于ALL、CLL患者(P<0.05),而LMO2的表达在正常人中明显低于ALL、CLL患者(P<0.05)。转染后,ALL、CLL患者中MicroRNA-223表达明显增加(571.86±142.00)(P<0.05),而LMO2表达明显减少(651.97±230.12)(P<0.05);在正常人中MicroRNA-223表达明显降低(646.32±172.93)(P<0.05),LMO2的表达明显升高(541.27±158.86)(P<0.05)。转染后细胞周期及细胞凋亡率的变化表现为,在ALL、CLL患者转染前细胞周期G1/G2细胞比例为(94.75±3.15)%,S期为(5.14±3.12)%;转染后G1/G2细胞比例明显增加(97.03±2.08)%(P<0.05),在S期明显减少(2.97±2.08)%(P<0.05);转染前细胞凋亡率为(54.47±8.72)%,转染后为(60.48±8.81)%,后者明显增加(P<0.05)。正常人转染前细胞周期G1/G2细胞比例是(96.73±2.26)%,S期是(3.25±2.26)%;转染后G1/G2细胞比例明显减少(94.55±2.77)%(P<0.05),在S期明显增加(5.45±2.77)%(P<0.05),细胞凋亡率为(59.02±10.20)%,转染后明显减少(51.96±10.20)%(P<0.05)。结论:淋巴细胞白血病原代细胞中MicroRNA-223表达降低,LMO2表达增高,导致淋巴细胞增殖周期及凋亡异�This study was aimed to investigate the expression level and mechanism of microRNA-223 and LM02 in acute lymphoblastic leukemia（ALL） and chronic lymphocytic leukemia（CLL） cells and the mechnism. MicroRNA-223 mimics was transfected to increase the expression of MicroRNA-223 in the lymphocytes sorted by ficoll separation from the bone marrow mononuclear cells （BMMNC） of ALL and CLL patients. MicroRNA-223 inhibitor was transfected to decrease the expression of the MicroRNA-223 in the lymphocytes of normal controls. Then the expression of the MicroR- NA-223 and LM02 in transfected lymphocytes before and after cultivating for 72 hours were detecfed by RT-PCR, the apoptosis and cell cycle of these cells were measured by flow cytometery. The results indicated that before the transfec tion, the expression of MicroRNA-223 in ALL and CLL cells was （433.11 ± 144.88）, which was significantly lower than that in the norinal lymphocyte（949.59 ± 267.39 ） ; the expression of LM02 was （ 807.10 ± 238.41 ）, which wassignificantly higher than that in the normal lymphocytes （455.32 ± 176.83 ） （P 〈 0.05 ） ; after the transfection, the expr- eseion of MicroRNA-223 was （571.86± 142.00） in ALL and CLL cells, which was significantly higher than that before transfection （P 〈0.05）, but the expression of LM02 was significantly lower than that before transfection （651.97±230.12 ） （ P 〈 0.05 ） ; in the normal control the expreseion of MicroRNA-223 obviously decreased （ 646.32 ± 172.93 ） （ P 〈 0.05 ）, the expression of LM02 was significantly increased （ 541.27 ± 158.86.2 ） （ P 〈 0.05 ）. After transfection, the cell cycle G1/G2 phase and apoptosis changed in ALL and CLL cells. Before transfection the cell ratio in cell cycle GiG2 phase was （94.75 ±3.15）%, the cell ratio in S phase was （5.14 ±3.12）% ; after transfection the cell ratio in cell cycle G/G2 phase was （97.03 ± 2.08 ）% and obviously increased（ P 〈 0.05 ）, the cell ratio in S phase was

关 键 词：急性淋巴细胞白血病 慢性淋巴细胞白血病 MicroRNA-223 LMO2基因 

分 类 号：R733.7[医药卫生—肿瘤]