Role of an autophagy/lysosome pathway in NF-κB pathway blocked pancreatic cancer Panc-1 cells

Ting Wu; Qi Yang; Miao Chen; Yizhong Feng

Role of an autophagy/lysosome pathway in NF-κB pathway blocked pancreatic cancer Panc-1 cells

Int J Clin Exp Pathol. 2020 Mar 1;13(3):437-446. eCollection 2020.

Authors

Ting Wu¹, Qi Yang², Miao Chen¹, Yizhong Feng³

Affiliations

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.

PMID: 32269680
PMCID: PMC7137030

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.

Keywords: Nuclear factor-κB; P53; SN50; autophagy.