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[摘要]
目的 研究花旗松素对人肺癌细胞A549增殖的抑制作用,并探讨其诱导细胞凋亡的可能机制。方法 WST-1法检测不同质量浓度(0~200 μg/mL)花旗松素对A549细胞增殖的影响;Annexin-V/PI双染法检测花旗松素对A549细胞凋亡的影响;细胞免疫荧光法观察花旗松素处理后A549细胞中凋亡蛋白Bax的表达;Western blotting法检测Bcl-2、Akt、P53蛋白表达的变化。结果 WST-1法检测到花旗松素能有效抑制A549细胞的增殖,呈现时间-剂量-效应关系。通过流式细胞仪分析,花旗松素诱导细胞凋亡,上调了促凋亡蛋白Bax的表达,下调抗凋亡蛋白Bcl-2的表达,抑制Akt的过表达,并且促进P53的表达。结论 花旗松素对A549细胞的增殖具有显著的抑制作用,其诱导的凋亡机制可能与Bcl-2、Akt相关信号通路的激活有关。
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[Abstract]
Objective To investigate the inhibition of taxifolin on proliferation of human lung cancer cell A549, and study the possible mechanism of inducing cell apoptosis. Methods WST-1 assay was employed to evaluate the inhibition of taxifolin with various concentrations from 0 μg/mL to 200 μg/mL on proliferation of A549 cells. With Annexin V and PI dual parameter markers, A549 cell apoptosis was detected by flow cytometry of taxifolin. The expressions of Bax, Bcl-2, Akt, and P53 were evaluated by Western blotting or immunocytochemistry. Results The WST-1 results showed that taxifolin could inhibit A549 cell viability with time-drug concentration-proliferation inhibition rate relationships. The results of Western blotting and fluorescence microscope showed that taxifolin could induce apoptosis of A549, up-regulate express of pro apoptotic protein Bax, down-regulate express of anti apoptotic protein Bcl-2, inhibit expression of Akt, and also induced the expression of P53. Conclusion Taxifolin can significantly inhibit the proliferation of A549 cell, and its mechanism may be related to activate the pathway of Bcl-2 and Akt-dependent apoptosis in A549 cells.
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