[Effects of Magnolol combined with Gefitinib on A549 non-small cell lung cancer cells].

Objective: To investigate the synergistic effects of magnolol and gefitinib on non-small cell lung cancer A549 cells. Methods: A549 cells were treated with Magnolol (6.25～500 μmol/L) or gefitinib (6.25～500 μmol/L) for 24 h, respectively, and the cell viability was detected by cell counting Kit-8 (CCK-8) experiment (n=3). Magnolol 100 μmol/L and gefitinib 5 μmol/L were selected in the following experiments (n=3, 24 h). Control group, magnolol group, gefitinib group and magnolol+gefitinib group were set up for factorial analysis. Colony formation experiment was applied to detect the cell proliferation. Western blot was used to detect protein expressions. Flow cytometry was applied to test cell apoptosis and sorting CD44+ and CD133+ cells. Results: Compared with the control group, the colony formation rate of Magnolol or Gefitinib groups was decreased significantly (P＜0.05); the apoptosis rate was increased significantly (P＜0.05); the number of CD44+ and CD133+ cells was reduced significantly (P＜0.05); the expressions of Ki67, PCNA, and stem cell marker proteins SOX2 and OCT4 were down-regulated (P＜0.05); and the ratio of Bax/Bcl-2 was increased significantly (P＜0.05). Compared with the Magnolol group or Gefitinib group, the Magnolol+Gefitinib group further promoted the above changes (P＜0.05), and the apoptosis rate, the ratio of Bax/Bcl-2, SOX2 and OCT4 all showed interactions between magnolol and gefitinib (P＜0.05). Conclusion: Magnolol and gefitinib promote the apoptosis of A549 cells and inhibit its stem cell-like properties, and the effect of the two combined is better than separated administration. Magnolol and gefitinib have interactive effects on A549 cells.目的: 探讨厚朴酚与吉非替尼协同影响非小细胞肺癌A549细胞的作用。方法: 以浓度为6.25～500 μmol/L厚朴酚、0.625～100 μmol/L吉非替尼分别处理A549细胞24 h,CCK-8实验检测细胞活力 (n=3),选24 h及100 μmol/L厚朴酚与5 μmol/L吉非替尼作后续处理(n=3);采用对照组、厚朴酚组、吉非替尼组和厚朴酚+吉非替尼组的析因分析设计;克隆形成检测细胞增殖;蛋白印迹测蛋白表达;流式细胞术检测细胞凋亡及分选CD44+和CD133+细胞。结果: 与对照组比,厚朴酚和吉非替尼组的克隆形成率显著降低(P＜0.05);凋亡率显著升高(P＜ 0.05);CD44+和CD133+细胞数量显著减少(P＜0.05);Ki67和PCNA及干细胞标记蛋白SOX2和OCT4表达显著下调(P＜0.05);Bax/Bcl-2表达比例显著上调(P＜0.05)。与厚朴酚组或吉非替尼组比较,厚朴酚+吉非替尼组进一步促进了上述改变(P＜0.05),且凋亡率、Bax/Bcl-2、SOX2和OCT4等指标都存在厚朴酚和吉非替尼的交互作用(P＜ 0.05)。结论: 厚朴酚与吉非替尼促进A549细胞凋亡和抑制其干细胞样特性,且联合用药效果优于单一给药。二者对A549细胞的抑癌作用有交互影响。.