An Liu1, Weiwei Zhang2, Yihui Chen2, Dawei Zhou2, Zhen Wang2, Jian Kang3, Le Wei4. 1. Department of Otorhinolaryngology, Third Xiangya Hospital, Central South University, Changsha, Hunan, China. 2. Department of Dermatology, Third Xiangya Hospital, Central South University, Changsha, Hunan, China. 3. Department of Dermatology, Third Xiangya Hospital, Central South University, Changsha, Hunan, China. Electronic address: xy3yyjiankang@126.com. 4. Department of Dermatology, Third Xiangya Hospital, Central South University, Changsha, Hunan, China. Electronic address: weile_xyzn@163.com.
Abstract
BACKGROUND: Increasing evidence has suggested that autophagy may play a resistant role during photodynamic therapy (PDT). The Wnt/β-catenin pathway is tightly involved in cell proliferation and autophagy. In this study, we aimed to determine the influence of 5-Ethylamino-9-diethylaminobenzo[a]phenoselenazinium (EtNBSe) mediated PDT (EtNBSe-PDT) on autophagy, proliferation and Wnt/β-catenin pathway in human NPC cell line (HNE-1 cells), and further explore the underlying crosstalk between them. METHODS: Cell viability and proliferation was evaluated by MTT assay. Autophagy and Wnt/β-catenin signaling pathway was analyzed by western blotting and immunofluorescence. RESULTS: It was revealed that EtNBSe-PDT significantly impeded the viability and proliferation of HNE-1 cells. Meanwhile EtNBSe-PDT could notably induce autophagy in HNE-1 cells accompanied with the inhibition of Wnt/β-catenin pathway. The Wnt/β-catenin pathway activator Wnt agonist was found to partially counteract the inhibitory proliferation of HNE-1 cells and suppress the autophagy induced by EtNBSe-PDT. In addition, pretreatment with the autophagy inhibitor 3-methyladenine (3-MA) or Wnt agonist showed the potential in enhancing the cytotoxic effect of EtNBSe-PDT (cell survival from 50.71 ± 4.16% to 24.53 ± 4.27% and from 52.64 ± 3.54% to 35.74 ± 4.27% respectively). CONCLUSION: Taken together, this study demonstrated that EtNBSe-PDT suppressed viability and proliferation, and induced autophagy of HNE-1 cells via downregulating the Wnt/β-catenin pathway. The autophagy further constituted the cytoprotective mechanisms involved in HNE-1 cells, which suggested that the combination of EtNBSe-PDT and autophagy inhibitors may be a promising strategy for the treatment of human NPC.
BACKGROUND: Increasing evidence has suggested that autophagy may play a resistant role during photodynamic therapy (PDT). The Wnt/β-catenin pathway is tightly involved in cell proliferation and autophagy. In this study, we aimed to determine the influence of 5-Ethylamino-9-diethylaminobenzo[a]phenoselenazinium (EtNBSe) mediated PDT (EtNBSe-PDT) on autophagy, proliferation and Wnt/β-catenin pathway in human NPC cell line (HNE-1 cells), and further explore the underlying crosstalk between them. METHODS: Cell viability and proliferation was evaluated by MTT assay. Autophagy and Wnt/β-catenin signaling pathway was analyzed by western blotting and immunofluorescence. RESULTS: It was revealed that EtNBSe-PDT significantly impeded the viability and proliferation of HNE-1 cells. Meanwhile EtNBSe-PDT could notably induce autophagy in HNE-1 cells accompanied with the inhibition of Wnt/β-catenin pathway. The Wnt/β-catenin pathway activator Wnt agonist was found to partially counteract the inhibitory proliferation of HNE-1 cells and suppress the autophagy induced by EtNBSe-PDT. In addition, pretreatment with the autophagy inhibitor 3-methyladenine (3-MA) or Wnt agonist showed the potential in enhancing the cytotoxic effect of EtNBSe-PDT (cell survival from 50.71 ± 4.16% to 24.53 ± 4.27% and from 52.64 ± 3.54% to 35.74 ± 4.27% respectively). CONCLUSION: Taken together, this study demonstrated that EtNBSe-PDT suppressed viability and proliferation, and induced autophagy of HNE-1 cells via downregulating the Wnt/β-catenin pathway. The autophagy further constituted the cytoprotective mechanisms involved in HNE-1 cells, which suggested that the combination of EtNBSe-PDT and autophagy inhibitors may be a promising strategy for the treatment of human NPC.