Role of an autophagy/lysosome pathway in NF-B pathway blocked pancreatic cancer Panc-1 cells
INTERNATIONAL JOURNAL OF CLINICAL AND EXPERIMENTAL PATHOLOGY(2020)
Abstract
Purpose: To investigate the role of an autophagy/lysosome pathway in NF-kappa B 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-kappa B was blocked by the inhibitor of p65 nuclear translocation, which activated autophagy and induced autophagic cell death in pancreatic cancer Panc-1 cell line.
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Key words
Nuclear factor-kappa B,SN50,autophagy,P53
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