茶多酚联合顺铂对肺癌细胞A549 增殖和凋亡的影响

Effect of tea ployphenols combined with cisplatin on A549 lung cancer cells and its mechanism

摘要:

目的 观察茶多酚（TP）、顺铂（DDP）及两者联合对人肺癌细胞A549 增殖和凋亡的影响，探 讨TP 联合DDP 联合影响A549 细胞生长的机制。方法 人肺癌细胞A549 经TP、DDP 单独或联合处理后， 将A549 细胞分成TP 组、DDP 组、TP 联合DDP 组（TP+DDP 组）及空白对照组，MTT 法检测细胞增殖 情况，Annexin V/PI 双染流式细胞术分析细胞凋亡，Western blot 检测细胞中p-AKT 和p-ERK1/2 的表达。 结果 TP 组、DDP 组及TP+DDP 组细胞增殖率分别为（91.0±3.6）%、（84.3±5.0）% 和（70.3±4.2）%， TP+DDP 组的细胞增殖率与DDP 组比较，差异有统计学意义（P <0.05），TP+DDP 组低于DDP 组；流式细胞 术示TP 组细胞凋亡率无明显影响，TP+DDP 组凋亡率与DDP 组比较，差异有统计学意义（P <0.05）；DDP 组细胞中p-AKT、p-ERK1/2 蛋白表达降低，TP 联合DDP 组中p-AKT、p-ERK1/2 的蛋白表达与DDP 组 比较，差异有统计学意义（P <0.05），TP+DDP 组低于DDP 组。结论 TP 与DDP 联合用药可增强DDP 的抑 制增殖和促进凋亡作用，其机制可能与下调AKT 的表达和ERK1/2 蛋白的磷酸化有关。

Abstract:

Objective To study the effect of tea ployphenols (TP), cisplatin (DDP) alone and in combination on the proliferation and apoptosis ability of human lung carcinoma cell line A549 in vitro and the mechanism. Methods A549 cells were treated with 10 μg/ml TP (TP group), 5 μg/ml DDP (DDP group) and 10 μg/ml TP plus 5 μg/ml DDP (TP+DDP group) respectively, the cells without treatment were used as the blank control group. Cell proliferation was determined by MTT assay. The morphology of apoptotic cells was observed by Annexin V/PI double staining. Flow cytometry was used to analyze apoptosis. The expressions of ERK1/2 and p-AKT were detected by Western blot. Results The cell proliferation rate of the TP, DDP and TP+DDP groups was (91.0 ± 3.6)%, (84.3 ± 5.0) % and (70.3 ± 4.2)% respectively. The anti-proliferation effect of the TP+DDP group was superior to that of the DDP group (P < 0.05). The results of flow cytometry showed that the apoptotic rate was not obviously changed in the TP group, but the apoptotic rate in the TP+DDP group was significantly different from that in the DDP group (P < 0.05). Low doses of TP had no significant apoptosis-promoting effect. Western blot analysis showed that the expressions of ERK1/2 and p-AKT decreased in the DDP group, which were significantly different from those in the TP+DDP group (P < 0.05).Conclusions TP combined with DDP can significantly inhibit the proliferation of A549 cells, induce cell apoptosis, which may be associated with the reduction of ERK1/2 expression and AKT phosphorylation.