白藜芦醇通过铁死亡途径逆转食管癌细胞Eca109/DDP化疗耐药

Resveratrol reverses drug resistance of esophageal cancer cell line Eca109/DDP via regulating ferroptosis

背景白藜芦醇不仅具有抗肿瘤作用,其还能增强多种化疗药物的化疗敏感性,但其对食管癌耐药细胞的顺铂敏感性的作用尚不清楚.目的探讨白藜芦醇逆转食管癌细胞Eca109/DDP的耐药效应并从铁死亡的角度分析其潜在的作用机制.方法采用MTT法确定白藜芦醇和顺铂的最佳作用时间和浓度;检测细胞增殖、细胞内丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)、活性氧(reactive oxygen species,ROS)及铁离子的水平;Western blot检测铁死亡相关调控因子酰基辅酶A合成酶长链家族成员4(acyl coenzyme A synthetase long chain family member 4,ACSL4)、铁蛋白重链(ferritin heavy chain,FTH)、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)和肿瘤蛋白53(tumor protein 53,P53)的蛋白表达情况.结果MTT检测结果显示,相较于单用顺铂,联用白藜芦醇对Eca109/DDP细胞的生长抑制作用较为显著(P<0.05).此外,白藜芦醇和顺铂联用可显著降低Eca109/DDP细胞克隆形成数(P<0.05),增加细胞内铁离子、MDA和ROS水平(P<0.05),降低GSH水平(P<0.05).铁死亡抑制剂(ferrostatin-1,Fer-1)和铁离子螯合剂(deferoxamine,DFO)可部分逆转白藜芦醇对细胞增殖的抑制作用(P<0.05).Western blot的结果显示相较于其他组,白藜芦醇和顺铂联用组的FTH和GPX4的蛋白表达显著降低(P<0.05),而ACSL4和P53的蛋白表达显著增加(P<0.05).结论白藜芦醇可通过铁死亡抑制Eca109/DDP细胞的增殖并逆转顺铂耐药.

BACKGROUND Resveratrol not only has anti-tumor effects,but also can enhance the chemosensitivity of tumor cells to a variety of chemotherapeutic agents.However,its effect on cisplatin sensitivity of drug-resistant esophageal cancer cells remains unclear.AIM To investigate whether resveratrol reverses the drug resistance of esophageal cancer Eca109/DDP cells and to explore the potential mechanism involved from the perspective of ferroptosis.METHODS The optimal treatment time and concentration of resveratrol and cisplatin were determined by MTT assay.Cell prolife-ration and the intracellular levels of malon-dialdehyde(MDA),glutathione(GSH),reactive oxygen species(ROS),and ferrous iron were detected.The protein expression of the ferroptosis-related molecules acyl coenzyme A synthetase long chain family member 4(ACSL4),ferritin heavy chain(FTH),glutathione peroxidase 4(GPX4),and tumor protein 53(P53)was detected by Western blot assay.RESULTS MTT assay showed that compared with cisplatin alone,resveratrol combined with cisplatin significantly inhibited the growth of Eca109/DDP cells(P<0.05).The combination of resveratrol and cisplatin not only reduced the number of colonies formed(P<0.05),but also increased the levels of ferrous iron,MDA,and ROS(P<0.05)and decreased the level of GSH(P<0.05)in Eca109/DDP cells.The inhibitory effect of resveratrol on Eca109/DDP cell proliferation was partially reversed by ferrostatin-1(Fer-1)and deferoxamine(DFO)(P<0.05).Western blot analysis showed that compared with other groups,the protein expression of FTH and GPX4 in the resveratrol and cisplatin combination group was significantly decreased(P<0.05),and the protein expression of ACSL4 and P53 was significantly increased(P<0.05).CONCLUSION Resveratrol can inhibit cell proliferation and reverse cisplatin resistance by regulating ferroptosis in Eca109/DDP cells.