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目的:研究染料木素(Genistein)对人骨髓瘤细胞株XG-1的核因子-κB(NF-κB)表达的影响及Genistein处理前后XG-1细胞内蛋白激酶B(Akt)和磷酸化Akt的表达。方法:凝胶电泳迁移率试验检测Genistein处理前后NF-κB在XG-1细胞核内的表达水平;蛋白质印迹法检测Genistein处理前后XG-1细胞内Akt和磷酸化Akt的表达。结果:凝胶电泳迁移率实验(EMSA)发现,NF-κB在XG-1细胞株中持续激活,而Genistein能下调这一激活。随着浓度逐渐提高,细胞核内NF-κB的活力逐渐下降,呈剂量依赖,其中溶剂对照组与10mg/L的Genistein组相比,χ2=4.614,P<0.05;溶剂对照组与15mg/L的Genistein组相比,χ2=4.324,P<0.05。Genistein能拮抗这一作用。XG-1细胞用不同浓度Genistein处理24h,可见Akt无明显变化,而磷酸化Akt随着浓度逐渐提高,表达逐渐下降,呈剂量依赖,其中溶剂对照组与10mg/L的Genistein组相比,χ2=4.751,P<0.05;溶剂对照组与15mg/L的Genistein组相比,χ2=5.114,P<0.05。结论:NF-κB在XG-1人类骨髓瘤细胞株中持续激活,Genistein下调NF-κB的激活形式而且抑制了Akt磷酸化,Genistein抑制细胞的NF-κB的激活部分经由Akt通路完成。
Objective: To investigate the effect of Genistein on the expression of nuclear factor-κB (NF-κB) in human myeloma cell line XG-1 and the effect of Genistein on Akt and phosphorylation of Akt expression. Methods: The expression of NF-κB in XG-1 cells was detected by gel electrophoretic mobility shift assay before and after Genistein treatment. The expression of Akt and phosphorylated Akt in XG-1 cells was detected by Western blotting. Results: Electrophoretic mobility shift assay (EMSA) showed that NF-κB was activated continuously in XG-1 cell line and Genistein down-regulated this activation. Compared with 10 mg / L Genistein group, χ2 = 4.614, P <0.05; solvent control group and 15 mg / L Compared with Genistein group, χ2 = 4.324, P <0.05. Genistein can antagonize this effect. XG-1 cells treated with different concentrations of Genistein for 24h, showing no significant changes in Akt, while the phosphorylated Akt with the gradual increase in concentration, the expression decreased gradually, in a dose-dependent manner, the solvent control group and 10mg / L Genistein group, χ2 = 4.751, P <0.05; χ2 = 5.114, P <0.05 compared with 15 mg / L Genistein group in solvent control group. CONCLUSION: NF-κB is activated continuously in XG-1 human myeloma cell line. Genistein down-regulates the activation of NF-κB and inhibits Akt phosphorylation. Activation of NF-κB by Genistein inhibits cell via Akt pathway.