Ting Wu1, Qi Yang2, Miao Chen1, Yizhong Feng3. 1. Department of Pathology, The Affiliated People's Hospital of Jiangsu University Zhenjiang 212000, Jiangsu Province, China. 2. Department of Pathology, Zhenjiang Hospital of Chinese Traditional and Western Medicine Zhenjiang 212002, Jiangsu Province, China. 3. Department of Pathology, The Second Affiliated Hospital of Soochow University Suzhou 215006, Jiangsu Province, China.
Abstract
PURPOSE: To investigate the role of an autophagy/lysosome pathway in NF-κB pathway blocked pancreatic cancer Panc-1 cells. METHODS: The inhibitory effects of SN50 on pancreatic cancer cell line Panc-1 were detected by MTT assay. After SN50 treatment, autophagy activation was observed by MDC staining and transmission electron microscope (TEM). The expression of light chain 3 (LC3) was detected by immunofluorescence staining. Western blotting analyses were used to detect the expression of apoptosis-related protein p53 and autophagy-related proteins LC3, p62, and Beclin1. RESULTS: Panc-1 cell activity was inhibited after SN50 treatment. The inhibition ratios of Panc-1 cells were (25.76±5.53)%, (34.35±4.49)% and (45.22±1.76)% after treatment of SN50 for 6 h, 12 h, and 24 h, and all changes were significant (P<0.05). Western blotting analysis showed that expressions of apoptotic protein p53, autophagic protein LC3, and Beclin 1 were increased, but the expression of p62 was down-regulated in Panc-1 cells. After SN50 treatment, immunofluorescence showed staining of microtubule-related protein 1 LC3, and MDC fluorescence staining showed increased autophagy bubbles labeled with MDC. Transmission electron microscope (TEM) was used to observe ultrastructure of Panc-1 cells that underwent autophagy after SN50 treatment. CONCLUSION: The activation of NF-κB was blocked by the inhibitor of p65 nuclear translocation, which activated autophagy and induced autophagic cell death in pancreatic cancer Panc-1 cell line. IJCEP
PURPOSE: To investigate the role of an autophagy/lysosome pathway in NF-κB pathway blocked pancreatic cancerPanc-1 cells. METHODS: The inhibitory effects of SN50 on pancreatic cancer cell line Panc-1 were detected by MTT assay. After SN50 treatment, autophagy activation was observed by MDC staining and transmission electron microscope (TEM). The expression of light chain 3 (LC3) was detected by immunofluorescence staining. Western blotting analyses were used to detect the expression of apoptosis-related protein p53 and autophagy-related proteins LC3, p62, and Beclin1. RESULTS:Panc-1 cell activity was inhibited after SN50 treatment. The inhibition ratios of Panc-1 cells were (25.76±5.53)%, (34.35±4.49)% and (45.22±1.76)% after treatment of SN50 for 6 h, 12 h, and 24 h, and all changes were significant (P<0.05). Western blotting analysis showed that expressions of apoptotic protein p53, autophagic protein LC3, and Beclin 1 were increased, but the expression of p62 was down-regulated in Panc-1 cells. After SN50 treatment, immunofluorescence showed staining of microtubule-related protein 1 LC3, and MDC fluorescence staining showed increased autophagy bubbles labeled with MDC. Transmission electron microscope (TEM) was used to observe ultrastructure of Panc-1 cells that underwent autophagy after SN50 treatment. CONCLUSION: The activation of NF-κB was blocked by the inhibitor of p65 nuclear translocation, which activated autophagy and induced autophagic cell death in pancreatic cancerPanc-1 cell line. IJCEP