Mangiferin抑制多发性骨髓瘤恶性生物学特性与发挥抗癌效应机制的实验研究

Experimental Study on the Mechanism of Mangiferin Inhibiting Malignant Biological Characteristics of Multiple Myeloma and Exerting Anticancer Effect

目的:探讨纯中药提取物Mangiferin对多发性骨髓瘤(MM)细胞恶性生物学行为的影响,分析Mangiferin抗骨髓瘤效应的分子机制,为MM替代治疗提供实验依据。方法:不同浓度Mangiferin干预人MM细胞株U266、RPMI8226细胞后,CCK-8法检测细胞增殖,Annexin V/PI双染流式细胞术检测细胞凋亡,Western blot检测凋亡及相关信号通路蛋白的表达,实时荧光定量聚合酶链式反应(qRT-PCR)检测基质金属蛋白酶(MMP)、CXC趋化因子受体(CXCR)家族的表达变化。结果:Mangiferin可抑制U266、RPMI8226细胞增殖活性,并诱导其凋亡。当Mangiferin干预U266和RPMI8226细胞48 h后,U266和RPMI8226细胞中Bcl-2家族促凋亡蛋白Bax表达上调,并下调survivin、Bcl-xL蛋白的表达同时水解活化caspase-3以促进细胞凋亡,且显著下调U266细胞中Bcl-2蛋白表达以诱导细胞凋亡(P<0.05)。在Mangiferin干预MM细胞后,其不仅可增加肿瘤抑制因子p53的表达水平,同时通过抑制抗凋亡分子的表达并下调AKT、NF-κB磷酸化水平继而诱发MM细胞程序性死亡。Mangiferin干预后可明显下调U266细胞CXCR4、MMP2及MMP9的表达(P<0.05),对CXCR2、CXCR7以及MMP13的表达基本无影响(P>0.05);而干预RPMI8226细胞后可下调CXCR4、MMP9、MMP13的表达(P<0.01),对MMP2表达影响微弱,对CXCR2、CXCR7的表达基本无影响(P>0.05)。结论:Mangiferin能抑制MM细胞增殖并诱导其凋亡,其机制可能与抑制NF-κB信号通路激活并通过影响Bcl-2家族蛋白表达以及使MMP、CXCR家族核心成员表达受抑等有关。

Objective:To investigate the effect of pure Chinese herbal extract Mangiferin on the malignant biological behaviors of multiple myeloma(MM) cells,and to analyze the molecular mechanism of the anti-myeloma effect of Mangiferin,so as to provide experimental basis for MM replacement therapy.Methods:U266 and RPMI8226 of human MM cell lines were intervened with different concentrations of Mangiferin.Cell proliferation was detected by CCK-8method.Annexin V/PI double staining flow cytometry was used to detect cell apoptosis.Western blot was used to detect the expression of apoptosis and related signaling pathway proteins,and real-time quantitative polymerase chain reaction(qRT-PCR) was used to detect the expression of matrix metalloproteinase(MMP) and CXC chemokine receptor(CXCR) family.Results: Mangiferin could inhibit the proliferation activity of U266 and RPM I8226 cells and induce cells apoptosis.After Mangiferin intervened in U266,RPM 18226 cells for 48 h,the expression of Bcl-2 family proapoptotic protein Bax was up-regulated,while the expression of survivin and Bcl-xL proteins was down-regulated and caspase-3 was hydrolyzed and activated to promote cell apoptosis,besides.the expression of Bcl-2 protein in U266 cells was also significantly down-regulated to induce apoptosis(P<0.05).After Mangiferin intervenes in MM cells,it can not only increase the expression level of tumor suppressor p53,but also induce programmed cell death of M M cells by inhibiting the expression of anti-apoptotic molecules and down-regulating the phosphorylation levels of AKT and NF-κB.In addition,after the intervention of Mangiferin,the expressions of CXCR4,MMP2 and MMP9 in U266 cells were down-regulated(P<0.05),while there is no effect on the expressions of CXCR2,CXCR7 and MMP13(P<0.05).However,the expressions of CXCR4,MMP9,and MMP13 in RPMI8226 cells were down-reg ulated(P<0.01),the expression of MMP2 was weakly affected,and the expression of CXCR2 and CXCR7 was basically not affected(P<0.05).Conclusion:Mangiferin can inhibit the proliferation and induce apoptosis of MM cells,and its mechanism may be related to inhibiting the activation of NF-κB signaling pathway,affecting the expression of Bcl-2 family proteins,and inhibiting the expression of core members of MMP and CXCR family.