目的 研究花旗松素對人肺癌細(xì)胞A549增殖的抑制作用,并探討其誘導(dǎo)細(xì)胞凋亡的可能機(jī)制。方法 WST-1法檢測不同質(zhì)量濃度(0～200 μg/mL)花旗松素對A549細(xì)胞增殖的影響;Annexin-V/PI雙染法檢測花旗松素對A549細(xì)胞凋亡的影響;細(xì)胞免疫熒光法觀察花旗松素處理后A549細(xì)胞中凋亡蛋白Bax的表達(dá);Western blotting法檢測Bcl-2、Akt、P53蛋白表達(dá)的變化。結(jié)果 WST-1法檢測到花旗松素能有效抑制A549細(xì)胞的增殖,呈現(xiàn)時間-劑量-效應(yīng)關(guān)系。通過流式細(xì)胞儀分析,花旗松素誘導(dǎo)細(xì)胞凋亡,上調(diào)了促凋亡蛋白Bax的表達(dá),下調(diào)抗凋亡蛋白Bcl-2的表達(dá),抑制Akt的過表達(dá),并且促進(jìn)P53的表達(dá)。結(jié)論 花旗松素對A549細(xì)胞的增殖具有顯著的抑制作用,其誘導(dǎo)的凋亡機(jī)制可能與Bcl-2、Akt相關(guān)信號通路的激活有關(guān)。

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.