槲皮素对人骨髓瘤细胞U266侵袭和迁移的影响

EFFECT OF QUERCETIN ON THE INVASION AND MIGRATION OF HUMAN MYELOMA U266 CELLS

目的研究槲皮素对骨髓瘤细胞U266侵袭和迁移的影响及机制。方法用不同浓度的槲皮素处理人骨髓瘤细胞U266,噻唑蓝(MTT)方法测定细胞存活变化,计算其半数抑制浓度。人骨髓瘤细胞U266分为空白对照组(不做药物处理)、阳性对照组(用地西他滨处理)、槲皮素组(半数抑制浓度的槲皮素处理)、PDTC组(NF-κB信号抑制剂PDTC处理)、PDTC+槲皮素组(半数抑制浓度的槲皮素和PDTC处理)共5组进行细胞培养。各组以MTT检测细胞存活能力,划痕愈合实验检测细胞运动能力,Transwell小室检测细胞侵袭和迁移能力,蛋白免疫印迹(Western blot)检测细胞中基质金属蛋白酶2(MMP-2)、核因子-κBp65亚型(NF-κBp65)和基质金属蛋白酶9(MMP-9)蛋白表达水平。结果槲皮素组、PDTC组、PDTC+槲皮素组细胞存活、侵袭、迁移能力及细胞中MMP-2、MMP-9、NF-κBp65表达水低于空白对照组(F=28.419~167.394,P<0.001)。PDTC+槲皮素组细胞存活、侵袭、迁移能力及细胞中MMP-2、MMP-9、NF-κBp65表达水平低于槲皮素组和PDTC组(P<0.05)。结论槲皮素能够通过抑制NF-κB信号通路降低骨髓瘤细胞的侵袭和迁移能力。

Objective To investigate the effect of quercetin on the invasion and migration of human myeloma U266 cells and related mechanisms.Methods Human myeloma U266 cells were treated with different concentrations of quercetin.MTT assay was used to measure the change in cell viability,and median inhibitory concentration was calculated.Human myeloma U266 cells were divided into blank control group(without treatment),positive control group(treated with decitabine),quercetin group(treated with quercetin at its median inhibitory concentration),pyrrolidine dithiocarbamate(PDTC)group(treated with the NF-κB signaling pathway inhibitor pyrrolidine dithiocarbamate),and PDTC+quercetin group(treated with quercetin at its median inhibitory concentration and PDTC)for cell culture.MTT assay was used to measure cell viability;the wound healing test was used to evaluate the invasion and migration abilities of cells;Western blot was used to measure the protein expression of matrix metalloproteinase-2(MMP-2),nuclear factor-kappa B(NF-κB)p65,and matrix metalloproteinase-9(MMP-9).Results Compared with the blank control group,the quercetin group,the PDTC group,and the PDTC+quercetin group had significantly lower cell viability,invasion and migration abilities,and expression of MMP-2,MMP-9,and NF-κBp65(F=28.419-167.394,P<0.001).Compared with the quercetin group and the PDTC group,the PDTC+quercetin group had significantly lower cell viability,invasion and migration abilities,and expression of MMP-2,MMP-9,and NF-κBp65(P<0.05).Conclusion Quercetin can reduce the invasion and migration of myeloma cells by inhibiting the NF-κB signaling pathway.