EGCG对肺癌A549/DDP细胞药物敏感性的影响及机制

EGCG Sensitisizes Cisplatin-resistant Human Lung Adenocarcinoma Cells A549/DDP to Cisplatin

目的探讨EGCG对肺癌耐药细胞株A549/DDP对顺铂敏感性的影响及作用机制。方法MTT法检测EGCG对A549/DDP细胞药物敏感性的影响。AO/EB染色法检测EGCG联合顺铂处理A549/DDP细胞前后凋亡的形态学改变;流式细胞术检测细胞凋亡率;Western Blot方法观察EGCG作用前后耐药蛋白GST-π、Survivin表达的变化。结果MTT比色法显示20、40、60μg/mL EGCG均可增加细胞对顺铂的敏感性;荧光显微镜下观察到EGCG处理后细胞体积缩小,核固缩,呈典型的凋亡形态学改变;Western Blot方法检测发现,不同浓度的EGCG处理细胞24 h后GST-π、Survivin蛋白表达均下调。流式细胞术显示,20、40、60μg/mL EGCG联合顺铂处理细胞24 h后,凋亡率逐渐增加,说明EGCG联合顺铂能够促进肺癌耐药细胞株A549/DDP凋亡。结论EGCG能够增加A549/DDP细胞对顺铂的敏感性,其机制可能与降低GST-π、Survivin蛋白表达从而诱导细胞凋亡有关。

Objective To investigate the effect of EGCG on apoptosis and drug sensitization in eisplatin - resistant human lung adenocarcinoma cell line A549/DDP and to determine the inhibitory effect of EGCG in cells A549/DDP. Methods Cytotoxicity of EGCG and sensibihty to doxo - rubicin （ DOX ） in A549/DDP cell hne, were studied by MTr assay. AO/EB method was used to display the apoptosis morphological of A_549/DDP cell induces by EGCG. The effect of EGCG on inducing of apoptosis was analyzed by flow cytometry （ FCM ）. Western blot analysis was performed to detect the expresssion of GST - π, Survivin protein after EGCG treatment. Results After 24 hours exposure to EGCG, partial cells presented the characteristic morphological changes of apoptosis under the light microscopy including cell shrinkage, nuclear condensation. Western Blot analysis showed that GST - π and Survivin were decreased, after to EGCG （ 20,40,60 μg/ml ）Analysis by flow cytometry showed that the percentages of apoptosis cells were increased. Conclusion EGCG can sensitize cells to DDP increase cell spontaneous, which might be relate to the performed to detect the expresssion of GST - π.Survivin, and could significantly induce apotosis by this regulation