白花蛇舌草乙醇提取物体外诱导人AML-M2白血病细胞凋亡机制研究

    Study on the Mechanism of Inducing Apoptosis of AML-M2 Cells in Vitro by Ethanol Extraction of Hedyotis Diffusa Willd.

    • 摘要: 目的 观察白花蛇舌草乙醇提取物(ethanol extract of Hedyotis diffusa Willd.,EEHDW)对AML-M2白血病Kasumi-1细胞增殖和凋亡的影响机制。方法 采用MTT比色法、Hoechest荧光染色检测测定EEHDW作用后Kasumi-1细胞增殖和凋亡的情况,Western blotting法检测Kasumi-1细胞中Bcl-2、Bax、caspase-3、caspase-9、Cyto-C、P65、p-P65、IkBα、p-IkBα、IKKα/β、C-myc以及AML1-ETO的蛋白水平。结果 EEHDW抑制急性髓系白血病Kasumi-1细胞增殖,并具有时间和浓度依赖性。0.04,0.06,0.08 mg·mL-1的EEHDW能诱导细胞凋亡(P<0.05或P<0.01)。EEHDW可以显著上调Cyto-C、cleaved PARP、cleaved caspase-3、cleaved caspase-9以及Bax的表达水平,而降低Bcl-2、C-myc和AML1-ETO蛋白的表达,同时EEHDW能够浓度依赖性地增加p-P65、p-IkBα的蛋白表达。结论 EEHDW诱导Kasumi-1细胞凋亡一方面是通过调节Bax/Bcl-2的表达影响线粒体途径,另外一方面还与激活NF-кB信号通路有关,在这个过程中同时抑制原癌基因C-myc和AML1-ETO融合基因的表达。

       

      Abstract: OBJECTIVE To observe the effect of ethanol extract of Hedyotis diffusa Willd. (EEHDW) on the proliferation and apoptosis of AML-M2 cells Kasumi-1 and its mechanism. METHODS MTT colorimetry and Hoechest staining were used to detect the effect of EEHDW on the proliferation and apoptosis of Kasumi-1 cells. Western blotting was used to detect the protein levels of Bcl-2, Bax, caspase-3, caspase-9, Cyto-C, p65, p-P65, IkBa, p-IkBa, IKKa/b, C-myc and AML1-ETO in Kasumi-1 cells. RESULTS EEHDW inhibited the proliferation of Kasumi-1 cells. The 0.04, 0.06 and 0.08 mg·mL-1 of EEHDW could induce apoptosis(P<0.05 or P<0.01). It could significantly up regulate the expression of Cyto-C, cleaved PARP, cleaved caspase-3, cleaved caspase-9 and Bax, but decreased the expression of Bcl-2, C-myc and AML1-ETO. Meanwhile, EEHDW could increase the expression of p-P65 and p-IKBα in a concentration dependent manner. CONCLUSION EEHDW induce apoptosis of Kasumi-1 cells not only related to the regulation of Bax/Bcl-2 expression, but also relate to the activation of NF-кB signal pathway, which inhibits the expression of C-myc and AML1-ETO fusion gene.

       

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