The expression and properties of alkaline phosphatase (ALP) in Eca109 cells, a cell line derived fromhuman esophageal cancer were studied with specific inhibition assay and polyacrylamide gel electrophoresis.The resul...The expression and properties of alkaline phosphatase (ALP) in Eca109 cells, a cell line derived fromhuman esophageal cancer were studied with specific inhibition assay and polyacrylamide gel electrophoresis.The results showed that ALP of Eca109 cells was heat stable and was strongly inhibited by L-pheuylalanine, but slightly inhibited by urea. Preduisolone could causedramatic increase in activity of ALP, but no change in ALP isozyme and concomitant increase in lactic dehydrogenase activity were found after prednisolone treatment. The results suggested that placental alkaline phosphatase as an oncodevelopmental gene product could be expressed ectopically by Eca109 cells and prednisolone could specifically induce increase in its activity.展开更多
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...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.展开更多
BACKGROUND The poor prognosis and rising incidence of esophageal cancer highlight the need for improved therapeutics that are essential prior to treatment.LCL161 is an SMAC(second mitochondrial activator of caspases)m...BACKGROUND The poor prognosis and rising incidence of esophageal cancer highlight the need for improved therapeutics that are essential prior to treatment.LCL161 is an SMAC(second mitochondrial activator of caspases)mimic and inhibitor of apoptosis protein(IAP)antagonist which exhibits anti-tumor effects and improves the chemical sensitivity of many cancers.AIM To ascertain the effects and mechanisms of the SMAC analog LCL161 on esophageal cancer cells.METHODS MTT assay and TUNEL assay were used to detect cell proliferation and apoptosis,respectively.Western blot analysis was used to study the molecular mechanisms of LCL161-induced death of ECA109 cells.RESULTS LCL161 decreased ECA109 cell proliferation in dose-and time-dependent manner and induced apoptosis of ECA109 cells in a dose-dependent manner.Also,LCL161 induced a significant decrease in the expression of the XIAP and significant increase in the expression of Caspase-3.In addition,Bax increased significantly with increasing concentrations of LCL161,and the relative expression of Bax was significantly different between groups.CONCLUSION These findings support the hypothesis that LCL161 can inhibit proliferation and induce apoptosis in esophageal cancer cells by regulating the expression of IAP family members,suggesting that it has potential to be an effective treatment for esophageal squamous cell carcinoma.展开更多
文摘The expression and properties of alkaline phosphatase (ALP) in Eca109 cells, a cell line derived fromhuman esophageal cancer were studied with specific inhibition assay and polyacrylamide gel electrophoresis.The results showed that ALP of Eca109 cells was heat stable and was strongly inhibited by L-pheuylalanine, but slightly inhibited by urea. Preduisolone could causedramatic increase in activity of ALP, but no change in ALP isozyme and concomitant increase in lactic dehydrogenase activity were found after prednisolone treatment. The results suggested that placental alkaline phosphatase as an oncodevelopmental gene product could be expressed ectopically by Eca109 cells and prednisolone could specifically induce increase in its activity.
文摘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.
文摘BACKGROUND The poor prognosis and rising incidence of esophageal cancer highlight the need for improved therapeutics that are essential prior to treatment.LCL161 is an SMAC(second mitochondrial activator of caspases)mimic and inhibitor of apoptosis protein(IAP)antagonist which exhibits anti-tumor effects and improves the chemical sensitivity of many cancers.AIM To ascertain the effects and mechanisms of the SMAC analog LCL161 on esophageal cancer cells.METHODS MTT assay and TUNEL assay were used to detect cell proliferation and apoptosis,respectively.Western blot analysis was used to study the molecular mechanisms of LCL161-induced death of ECA109 cells.RESULTS LCL161 decreased ECA109 cell proliferation in dose-and time-dependent manner and induced apoptosis of ECA109 cells in a dose-dependent manner.Also,LCL161 induced a significant decrease in the expression of the XIAP and significant increase in the expression of Caspase-3.In addition,Bax increased significantly with increasing concentrations of LCL161,and the relative expression of Bax was significantly different between groups.CONCLUSION These findings support the hypothesis that LCL161 can inhibit proliferation and induce apoptosis in esophageal cancer cells by regulating the expression of IAP family members,suggesting that it has potential to be an effective treatment for esophageal squamous cell carcinoma.
