中文摘要：

目的研究miR-202对多发性骨髓瘤U266细胞中B细胞活化因子（BAFF）的调节作用及其机制。方法通过生物信息学软件预测miR-202与BAFF的潜在结合位点，构建荧光素酶报告基因载体。将人类miR-202—3P模拟物（has—miR-202—3P—mimics）、人类miR-202—3P抑制物（has—miR-202，inhibitor）、siBAFF及各自阴性对照转染U266细胞，采用实时荧光定量PCR和Westernblot法验证miR-202对BAFF的调控作用，采用细胞增殖实验和流式细胞仪检测转染后U266细胞的增殖能力和凋亡情况。结果未转染组、has—miR-202-3P—mimics转染组、has—miR-202—3P-inhibitor转染组和siBAFF转染组U266细胞中BAFFmRNA的相对表达量分别为1．0404-0．057、0．5734-0．073、1．205±0．097和0．368±0．052。与未转染组比较，has—miR-202-3P—mimics转染组和siBAFF转染组U266细胞中BAFFmRNA的表达明显下调（P〈0．05）。Westernblot法检测BAFF蛋白的表达结果与mRNA基本一致。未转染组、has．miR-202．3P．mimics转染组、has—miR-202-3P—inhibitor转染组和siBAFF转染组U266细胞的A450。。值分别为1．063±0．052、0．7144-0．045、0．9364-0．066和0．764±0．053。与未转染组比较，has—miR-202—3P-mimics转染组和siBAFF转染组U266细胞的A450nm值明显降低（P〈0．05）。未转染组、has-miR-202-3P-mimics转染组、has—miR-202-3P—inhibitor转染组和siBAFF转染组U266细胞的凋亡率分别为26．2％、49．6％、21．1％和30．7％。与未转染组比较，has．miR-202．3P—mimics转染组U266细胞的凋亡率明显增加（P〈0．05）。has—miR-202-3P—mimics转染组U266细胞中p-JNK蛋白的表达水平降低。结论miR-202通过调节BAFF的表达对U266细胞发挥抑制增殖和诱导凋亡的作用。JNK／SAPK信号通路参与了miR-202对BAFF的调控作用。

英文摘要：

Objective To explore the regulating effect of miR-202 on B cell-activating factor, and check whether the regulation influences the growth of multiple myeloma cells. Methods The potential binding sites of BAFF for miR-202 were predicted using bioinformatics software. Luciferase reporter gene analysis was used to evaluate the regulatory effect of miR-202 on BAFF. Human multiple myeloma U266 cells were transfected with has-miR-202-mimics, has-miR-202-inhibitor, siBAFF and their negative controls, respectively. After above treatments, BAFF mRNA and protein levels were detected by real-time PCR and Western blot analysis, and the proliferation and apoptosis in the multiple myeloma （MM） cells were examined by WST-1 and annexin V-FLUOS assay, respectively. Results The BAFF mRNA expression levels in the untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P- inhibitor transfeeted group and siBAFF transfected group were 1. 040：1： 0. 057, 0. 573 ±0. 073, 1. 205 ±0. 097 and 0. 368 ＋ 0. 052, respectively. BAFF mRNA expressions in U266 cells transfected with has-miR- 202-3P-mimics and siBAFF were significantly decreased compared with that in the untransfeeted group （ P 〈 0.05）. The BAFF protein expression level of each group was consistent with the mRNA assay result. The absorbanee value in 450 nm of the untransfeeted group, has-miR-202-3P-mimics transfected group, hasmiR-202-3P-inhibitor transfected group and siBAFF transfected group were 1. 063 ± 0.052, 0.714 ± 0.045, 0.936± 0. 066 and 0.764 ±0.053, respectively. In comparison with the untransfected group, the absorbance value at 450 nm of has-miR-202-3P-mimics and siBAFF transfected groups was significantly reduced （P 〈 0.05 ）. The cell apoptosis rates of untransfected group, has-miR-202-3P-mimies transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 26.2% , 49.6%, 21.1% and 30.7%, respectively. Therefore, the cell apoptosis rate of has-miR-202-3P-mimics transfected group was signifi