KRAS-driven pancreatic ductal adenocarcinoma cell lines harbour putative cancer stem cells

Objectives: Pancreatic cancer stem cells (CSCs) are known culprits of recurrent pancreatic ductal adenocarcinoma (PDAC). The disputable presence of CSCs in continuous cell lines has led to an extensive application of patient-derived CSC culture for experimentation. However, obtaining patient biopsies remains a challenge for many laboratories. Here, this study aimed to investigate the presence of CSCs in KRAS-driven PDAC cell lines. Methods: Cell viability assays evaluated the cytotoxicity of gemcitabine and vismodegib in PDAC cell lines. Flow cytometry was used to analyse the expression of pancreatic CSC surface markers. Tumoursphere-forming ability was investigated by culturing cells in serum-free and non-adherent conditions in vitro, while the tumourigenicity of PDAC cells was assessed in immunocompromised mice. Molecular events were delineated by Western blotting, and SOX2-expressing cells in tumour xenografts were detected by immunohistochemistry. Results: PANC-1 and Capan-2 cell lines showed exceptional chemoresistance, with Capan-2 containing a notable number of putative CSCs that co-expressed CD44, CD24, and CD133 (3.7 % of the total population) and sustained the cytotoxicity of gemcitabine. The Capan-2 cell line harbours tumoursphere-forming cells, of which self-renewal was inhibited by foetal bovine serum. The presence of these cells is consistent with Capan-2 being highly tumourigenic in immunocompromised mice. MAPK, PI3K-AKT, NF-κB, and WNT pathways were activated, and SOX2 expression was upregulated during the self-renewal of putative CSCs. SOX2-expressing cells were correspondingly present in xenografts of highly tumourigenic Capan-2 cell lines. Conclusion: The Capan-2 cell line contains a subset of putative CSCs, whose self-renewal is driven by MAPK, PI3K-AKT, NF-κB, and WNT pathways.

Text 124734.pdf - Published Version Available under License Creative Commons Attribution. Download (1MB) Official URL or Download Paper: https://www.degruyterbrill.com/document/doi/10.151...

Actions (login required)

View Item