Effect Of Chalcone Derivative Cc-85 On The Apoptosis Of Esophageal Cancer Cells Induced By Reactive Oxygen Species

PurposeThe anti-tumor activity of cc-85 was studied by investigating the mechanism of the new chalcone derivative cc-85 in esophageal cancer cell lines.The anti-tumor activity of cc-85 was studied through the death receptor pathway and mitochondrial pathway,aiming at Nrf2 as the target.It provides the theoretical basis for the development of tumor drug.MethodsWe studied contents of the subject through a series of pharmacology and molecular biology experimental methods.Cell viability was detected by MTT assay,the level of reactive oxygen species and the mitochondrial membrane potential were detected by flow cytometry.Protein expression was detected by western blot.DAPI method was used to detect the nuclear morphology and Immunofluorescence assay was used to detect protein localization.ResultsCc-85 inhibited cell viability of esophageal cancer cell lines,such as EC109,KYSE750 and EC-1.The results showed that cc-85 had a relatively small toxicity to normal esophageal epithelial cells and it was more sensitive to EC109 and KYSE750;cc-85 could induce the apoptosis of EC109 and KYSE750 cells;DAPI staining showed that the nuclear morphology of EC109 cells was smaller,the staining was deeper than control and some apoptotic bodies were observed;expressions of DR4 and DR5 in KYSE750 cells were significantly up-regulated by cc-85;cc-85 could decrease mitochondrial membrane potential,then regulate expressions of intracellular anti-apoptotic protein and pro-apoptotic protein;cc-85 could activate caspase-3,8,9,and caspase full enzyme inhibitor z-VAD-fmk could partially block the apoptosis which cc-85 induced;NAC,ROS inhibitor,could reverse the apoptosis of EC109 and KYSE750 cells induced by cc-85;NAC reversed the mitochondrial membrane potential of EC109 cells induced by cc-85;NAC reversed expressions ofanti-apoptotic protein and pro-apoptotic protein in EC109 cells induced by cc-85;NAC reversed protein expressions of the caspase family in EC109 cells induced by cc-85;cc-85 induced Nrf2 localization to the nucleus.ConclusionsAll of the above results demonstrated that chalcone derivative cc-85 can induce the apoptosis of esophageal cancer cells.Cc-85 could promote Nrf2 into the nuclear transcription,increase expressions of death receptors DR4 and DR5,decrease the mitochondrial membrane potential,regulate pro-apoptotic proteins,and induce other series of cell signaling pathways of apoptosis.All of these could provide important theoretical basis for clinical.