青蒿琥酯逆转食管癌Eca109/ADM细胞对多柔比星的耐药

Role of artesunate in resistance-reversal of esophageal cancer Eca109/ADM cells to doxorubicin

目的:研究青蒿琥酯(artesunate,Art)逆转食管癌细胞Eca109/ADM对多柔比星(doxorubicin,ADM)的耐药作用及其机制。方法:实验分为生理盐水(normal saline,NS)对照组(NS组)、Art组(0.1μmol/L)、ADM组(0.2μg/ml)和Art+ADM联合组。Art、ADM、Art+ADM作用Eca109/ADM细胞48 h后,流式细胞术检测细胞凋亡率、细胞内ADM的含量及细胞中三磷酸腺苷结合转运蛋白G超家族成员2(ATP-binding cassette transporter G2,ABCG2)蛋白的表达量,Western blotting检测细胞中ABCG2蛋白表达水平。结果:Art+ADM作用Eca109/ADM细胞48 h后,细胞的凋亡率[(12.89±0.87)%]显著高于Art组[(1.58±0.12)%]、ADM组[(6.55±0.90)%]及NS组[(1.44±0.10)%](P<0.05),Art可提高Eca109/ADM细胞对ADM的敏感性。流式细胞术检测结果显示,Art+ADM组Eca109/ADM细胞中ABCG2蛋白表达量(644.60±3.21)显著低于ADM组(659.15±4.59)及NS组(658.14±6.88)(P<0.05),但与Art组(644.31±3.96)相比无显著差异(P>0.05)。Western blotting检测结果与流式细胞术结果一致,Art组Eca109/ADM细胞中ABCG2蛋白的表达水平为(0.70±0.02),与对照组的(0.80±0.03)相比显著降低(P<0.05),Art+ADM组的ABCG2蛋白(0.71±0.04)与单独应用ADM组的ABCG2蛋白(0.81±0.05)相比,ABCG2蛋白表达水平显著降低(P<0.05)。Art+ADM组Eca109/ADM细胞内ADM的含量(848.02±5.04)显著高于单独应用Art组(763.29±4.02)、ADM组(800.25±3.84)及NS组(763.88±2.03)(P<0.01)。结论:Art可以降低食管癌Eca109/ADM细胞内ABCG2蛋白表达,增加ADM的含量,逆转肿瘤细胞对ADM的耐药。

Objective： To explore the role of artesunate（Art） in resistance-reversal of Eca109/ADM cells and the related mechanism.Methods：The present study was divided into 4 groups： normal saline（NS） control group,Art（0.1 μmol/L） group,ADM（0.2 μg/ml） group,and Art（0.1 μmol/L）＋ADM（0.2 μg/ml） group.The apoptosis rate,ADM content and ABCG2（ATP-binding cassette transporter G2） protein expression in Eca109/ADM cells were detected by flow cytometry 48 h after treated with Art,ADM,and Art＋ADM.ABCG2 protein expression in Eca109/ADM cells was further examined by Western blotting analysis.Results：The apoptosis rate of Eca109/ADM cells in Art＋ADM group was（12.89±0.87）%,being significantly higher than that in the Art group（1.58±0.12）%,ADM group（6.55±0.90）% and NS group（1.44±0.10）%（P〈0.05）.Art increased the sensitivity of Eca109/ADM cells to ADM.Flow cytometry results showed that the ABCG2 protein expression in Eca109/ADM cells of Art＋ADM group（644.60±3.21） was significantly lower than that in ADM group（659.15±4.59） and NS group（658.14±6.88）（P〈0.05）,but was similar to that in Art group（644.31±3.96）（P〉0.05）.Western blotting analysis results were consistent with those detected by flow cytometry.The ABCG2 protein expression of Eca109/ADM cells in Art group（0.70±0.02） was significantly decreased compared with NS group（0.80±0.03）（P〈0.05）,and that in Art＋ADM group（0.71±0.04） was also decreased compared with ADM group（0.81±0.05）（P〈0.05）.The ADM content of Eca109/ADM cells in Art＋ADM group was significantly higher than those in Art,ADM and NS groups（848.02±5.04 vs 763.29±4.02,800.25±3.84,763.88±2.03;P〈0.01）.Conclusion： Art can decrease ABCG2 protein expression and increase ADM content in Eca109/ADM cells,and it can also reverse the drug resistance of Eca109/ADM cells.