紫背万年青提取物联合吉非替尼对A549肺癌细胞的协同作用研究

Synergistic Inhibitory Effect of Extract From Rhoeo Discolor and Gefitinib on Human Lung Adenocarcinoma A549 Cells

目的:探讨紫背万年青提取物与吉非替尼联用对A549肺癌细胞的协同作用,并阐明其可能的分子机制。方法:采用CCK-8法检测细胞活力和细胞增殖的影响,并通过Isobologram方法进行联用效应分析。采用流式细胞术检测细胞凋亡和细胞周期。采用Western blot法检测相关蛋白,包括Bcl-2、Bax、p-PI3K和p-Akt的表达水平。结果:与单用吉非替尼相比,紫背万年青提取物与吉非替尼联用能显著降低吉非替尼抑制A549细胞IC50(P<0.05);Isobologram分析表明联用具有协同效应。联用可以显著诱导细胞的凋亡(P<0.05),同时上调凋亡标记蛋白Bax/Bcl-2的表达比例(P<0.05)。同时,联用也可以显著提高G0/G1期的比例(P<0.05),促进细胞分裂G0/G1停滞。Western blot结果表明,联用可以显著下调p-PI3K和p-Akt的蛋白表达水平(P<0.05)。结论:紫背万年青提取物与吉非替尼联用可协同抑制细胞增殖、阻滞细胞周期和促进细胞凋亡,其机制可能与抑制PI3K/Akt信号通路有关。

Objective:To investigate synergistic effects of extract from Rhoeo discolor(Water extract from Rhoeo discolor,WER)combined with gefitinib(GEF)on A549 lung cancer cells,and to explore underlying mechanisms.Methods:Cell viability and cell proliferation were detected by CCK-8 analysis,and the combination effect was analyzed by Isobologram analysis.Cell apoptosis and cell cycle were detected by flow cytometry test.Western blot was used to detect the expression levels of related proteins,including Bcl-2,Bax,p-PI3K and p-Akt.Results:Compared with GEF-treated alone,the combination of WER and GEF significantly reduced the IC50 of GEF;Isobologram analysis showed that the combination of WER and GEF provided a synergistic effect in A549 cells.The combination of WER and GEF could significantly enhance cell apoptosis and up-regulate the expression of apoptosis marker protein,Bax/Bcl-2.In addition,the combination could also significantly increase the ratio of G0/G1 phase and promote cell division G0/G1 stagnation.Western blot indicated that the combination could significantly down-regulate the protein expression of p-PI3K and p-Akt.Conclusion:The combination of WER and GEF could synergistically inhibit cell proliferation,block cell cycle and promote cell apoptosis,and the mechanism might be well associated with the inhibition of PI3K/Akt signaling pathway.