论文部分内容阅读
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。
The purpose of this study was to investigate the expression of MicroRNA-223 and LMO2 in primary lymphocytes and chronic lymphocytic leukemia (CLL) primary cells and the mechanism of action of MicroRNA-223. The human lymphocyte subsets of ALL, CLL patients and normal human bone marrow lymphocytes were selected, and MicroRNA-223 analogs in bone marrow lymphocytes of ALL and CLL patients were targeted to enhance the expression of MicroRNA-223 MicroRNA-223 expression in target-knockdown cells transfected with MicroRNA-223 inhibitor in normal human bone marrow lymphocytes. The expression of MicroRNA-223 and LMO2 before and after transfection was detected by RT-PCR and the correlation between the expression of MicroRNA-223 and LMO2 was observed after 72 h of transfection. The changes of apoptosis and cell cycle were further observed by flow cytometry. The results showed that the expression of MicroRNA-223 in ALL and CLL patients was (433.11 ± 144.88) and the level of LMO2 was (807.10 ± 238.41) before transfection with MicroRNA-223 analogs. Before microRNA-223 inhibitor transfection, The expression level of -223 was (949.59 ± 267.39) and LMO2 was (455.32 ± 176.83). The expression of MicrRNA-223 in normal controls was significantly higher than that in ALL and CLL patients (P <0.05) Was significantly lower than ALL, CLL patients (P <0.05). After transfection, the expression of MicroRNA-223 in ALL and CLL patients was significantly increased (571.86 ± 142.00) (P <0.05), while the expression of LMO2 was significantly decreased (651.97 ± 230.12) (P <0.05) (646.32 ± 172.93) (P <0.05), and the expression of LMO2 increased significantly (541.27 ± 158.86) (P <0.05). The changes of cell cycle and apoptosis rate after transfection showed that the percentage of G1 / G2 cell cycle was (94.75 ± 3.15)% in SLE and (5.14 ± 3.12)% in SLL, The percentage of G1 / G2 cells was significantly increased (97.03 ± 2.08)% (P <0.05) in S phase and significantly decreased (2.97 ± 2.08)% in S phase (54.47 ± 8.72) %, After transfection (60.48 ± 8.81)%, the latter was significantly increased (P <0.05). The percentage of G1 / G2 cells was (96.73 ± 2.26)% and that of S phase was (3.25 ± 2.26)%, respectively. The percentage of G1 / G2 cells decreased significantly after transfection (94.55 ± 2.77)% ) Significantly increased in S phase (5.45 ± 2.77)% (P <0.05). The apoptosis rate was (59.02 ± 10.20)% in S phase and decreased significantly (51.96 ± 10.20)% after transfection (P <0.05). Conclusion: The expression of MicroRNA-223 in primary lymphoblastic leukemia cells is decreased and the expression of LMO2 is increased, leading to lymphocyte proliferation cycle and abnormal apoptosis, which may be one of the pathogenesis of lymphocytic leukemia.