4′,5,7-三羟基异黄酮抗骨髓瘤细胞增殖机制的研究

Inhibitory effect of genistein on the proliferation of multiple myeloma cell

目的:为了研究4′,5,7-三羟基异黄酮(genistein,Gen)是否能抑制人多发性骨髓瘤(multipule myeloma,MM)细胞株XG1的增殖,研究Gen处理骨髓瘤细胞后B细胞淋巴瘤/白血病-2基因(bcl-2)、bcl-xl、细胞周期蛋白D1(cyclin D1)、细胞间黏附因子1(ICMA-1)基因表达变化及骨髓瘤细胞中核转录因子κB(NF-κB)表达的变化。方法:利用体外培养人MM细胞株XG1,依据剂量梯度分别给予Gen,用噻唑蓝染色法(MTT)观察不同药物浓度的Gen对MM细胞的增殖影响;5、10、15μg/mLGen与溶剂对照组分别作用XG1细胞48h后,半定量逆转录聚合酶链反应(RT-PCR)法检测XG1细胞bcl-2、bcl-xl、细胞周期蛋白D1、ICMA-1基因表达;应用5μg/mLGen处理XG1细胞,用免疫组织化学法观察Gen处理前后细胞内NF-κB的表达情况。结果:Gen作用XG1细胞48h,随浓度增加,细胞增殖逐渐下降;5、10、15μg/mLGen处理组mRNA的表达均低于溶剂对照组(P<0.05),伴随Gen处理浓度逐渐增加,bcl-2、bcl-xl、细胞周期蛋白D1、ICMA-1基因mRNA的表达逐渐下降;Gen能够使NF-κB在XG1细胞内重新分布,胞浆内出现NF-κB表达,胞核内NF-κB表达下降。结论:Gen可能通过下调骨髓瘤细胞核中NF-κB的表达来抑制bcl-2、bcl-xl、细胞周期蛋白D1、ICMA-1基因mRNA的表达,进而抑制骨髓瘤细胞的增殖、黏附、转移。

Objective To investigate the inhibitory effect of genistein on the proliferation of human multiple myeloma （MM） cell and the explession of B-cell leukemia-lymphoma bcl-2, bcl-xl, cyclin D1, ICMA-1 gene expression and the change of expression of nuclear transcription factor （NF）-κB after treated the myeloma cell with genistein. Methods Human MM cell line XG1 cells were treated with varying doses of genistien. The effect of genistein on the proliferation of XG1 cells was determined by 3-（4,5-dimethyhhiazol-2-yl）-2,5-diphenyhetrazolium bromide（MTF） assay. Gene expressions of bcl-2, bcl-xl, cyclin D1, ICMA-1 were measured by reverse transcriptase-polymerase chain reaction （RT-PCR）. Intracellular expression of NF-κB was assayed by immunohistochemistry. Results The proliferation of XG1 cell decreased dose dependently after treated with genistein for 48 hours. Expressions of bcl-2, bcl-xl, cyclin D1 and ICMA-1 mRNA decreased in a dose dependant manner after treated with 5, 10, 15 μg/mL genlstein. Genistein caused redistribution of NF- κB, with the expression of NF-κB in cytoplasm and decrease of NF-κB expression in nucleus. Conclusions Genistein inhibited the expressions of bcl-2, bcl-xl, cyclin D1, ICMA-1 mRNA via down-regulating the intranuclear expression of NF-κB, thereby inhibiting the proliferation, adhesion and metastasis of MM cells.