【摘要】 目的 探讨miRNA-106a在人淋巴瘤Jurkat细胞中的表达及作用机制。方法 取对数生长期人淋巴瘤Jurkat细胞，分别向培养基中加入5 ml生理盐水配制的浓度为0、0.5、1.0、1.5μg/ml的多柔比星，取健康体检者（对照组）的单个核细胞。采用定量逆转录聚合酶链反应（qRT-PCR）检测miRNA-106a以及视网膜母细胞瘤1(RB1)、E2F转录因子1(E2F1)、胱天蛋白酶3(caspase 3)mRNA的表达水平，采用噻唑蓝（MTT）法检测细胞增殖能力，流式细胞术检测细胞凋亡能力，采用蛋白质印迹法（Western blot）检测RB1、E2F1、caspase 3蛋白的表达水平。结果 0μg/ml多柔比星干预人淋巴瘤Jurkat细胞miRNA-106a的表达水平明显高于对照组单个核细胞（P﹤0.01）。随多柔比星浓度升高、作用时间延长，miRNA-106a表达水平逐渐降低，光密度（OD）值逐渐降低，细胞增殖抑制率（IR）和凋亡率均逐渐升高，RB1、caspase 3 mRNA及其蛋白的表达水平均逐渐升高，E2F1 mRNA及其蛋白的表达水平均逐渐降低，差异均有统计学意义（P﹤0.05）。相关性分析结果显示，miRNA-106a与RB1、caspase 3的表达均呈负相关（P﹤0.01），与E2F1的表达呈正相关（P﹤0.01）。结论 miRNA-106a在人淋巴瘤Jurkat细胞中高表达，其可能通过调控RB/E2F1通路相关蛋白的表达来调节淋巴瘤进展。更多还原

【Abstract】 Objective To study the expression and mechanism of miRNA-106a in human lymphoma Jurkat cells.Method Take human lymphoma Jurkat cells in logarithmic growth phase, and prepare 5 ml of physiological saline in the culture medium with concentrations of 0, 0.5, 1.0, and 1.5 μg/ml of doxorubicin. Take blood mononuclear cells from healthy individuals(control group). Quantitative reverse transcription-polymerase chain reaction(qRT-PCR) was used to detect the expression levels of miRNA-106a and retinoblastoma 1(RB1), E2F transcription factor 1(E2F1), and caspase3 mRNA. Methyl thiazolyl terazolium(MTT) assay was used to detect cell proliferation ability, flow cytometry was used to detect cell apoptosis ability, and Western blot was used to detect the expression levels of RB1, E2F1, and caspase 3protein. Result The expression levels of miRNA-106a in Jurkat cells of lymphoma treated with 0 μg/ml doxorubicin were higher than that in blood mononuclear cells from control group(P<0.01). As the concentration of doxorubicin increased and the duration of action prolonged, the expression levels of miRNA-106a gradually decreased, the optical density(OD) gradually decreased, and the inhibition rate(IR) of cell proliferation and apoptosis gradually increased, the expression levels of RB1, caspase 3 mRNA and their proteins also gradually increased, while the expression levels of E2F1mRNA and its protein gradually decreased, and the differences were statistically significant(P<0.05). The correlation analysis results showed that miRNA-106a were negatively correlated with the expression of RB1 and caspase 3(P<0.01), and positively correlated with the expression of E2F1(P<0.01). Conclusion miRNA-106a is highly expressed in human lymphoma Jurkat cells, and may regulate lymphoma progression by regulating the expression of RB/E2F1 pathway related proteins.更多还原