【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常,这可能是淋巴细胞白血病的发病机制之一。更多还原

【Abstract】 This study was aimed to investigate the expression level and mechanism of microRNA-223 and LMO2 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 MicroRNA-223 and LMO2 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 transfection,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 LMO2 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 expreseion 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 LMO2 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 LMO2 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 G₁/G₂ 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 G₁/G₂ phase was（97.03±2.08）% and obviously increased（P<0.05）,the cell ratio in S phase was（2.97±2.08）% and significantly decreased（P<0.05）.Before transfection the apoptosis rate was （54.47±8.72）%,and obviously was higher than that after transfection（60.48±8.81）%.And in the normal control,the cell ratio in G₁/G₂ phase was significantly higher than that after transfection [（96.73±2.26）%,（94.55±2.77）%,P<0.05)],and the cell ratio in S phase was significantly increased[（3.25±2.26）%,（5.45±2.77）%（P<0.05）].The apoptotic rate in the ALL and CLL patients was significantly higher than that after the transfection [（54.47±8.72）% vs（60.48±8.81）%,respectively（P<0.05）].The apoptotic rate in the narnol control was significantly lower than that after the transfection[（59.02±10.20）%,（51.96±10.20）%,respectively（P<0.05）].It is concluded that the expression of MicroRNA-223 decreases,and the expression of LMO2 inceases in lymphocytic leukemia cells which leads to the lymphocytes over-proliferation and abnormal apoptosis,thus may be one of pathogenesis in lymphocytic leukemia.更多还原