Pancreatic stellate cell-induced gemcitabine resistance in pancreatic cancer is associated with LDHA- and MCT4-mediated enhanced glycolysis

Manoj Amrutkar; Kjersti Berg; Aina Balto; Miguel G Skilbrei; Anette V Finstadsveen; Monica Aasrum; Ivar P Gladhaug; Caroline S Verbeke

doi:10.1186/s12935-023-02852-7

Pancreatic stellate cell-induced gemcitabine resistance in pancreatic cancer is associated with LDHA- and MCT4-mediated enhanced glycolysis

Cancer Cell Int. 2023 Jan 19;23(1):9. doi: 10.1186/s12935-023-02852-7.

Authors

Manoj Amrutkar^{1

2}, Kjersti Berg³, Aina Balto³, Miguel G Skilbrei³, Anette V Finstadsveen³, Monica Aasrum⁴, Ivar P Gladhaug^{5

6}, Caroline S Verbeke^{3

7}

Affiliations

¹ Department of Pathology, Oslo University Hospital Rikshospitalet, Oslo, Norway. manoj.amrutkar@medisin.uio.no.
² Department of Pharmacology, Institute of Clinical Medicine, University of Oslo, Oslo, Norway. manoj.amrutkar@medisin.uio.no.
³ Department of Pathology, Oslo University Hospital Rikshospitalet, Oslo, Norway.
⁴ Department of Pharmacology, Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
⁵ Department of Hepato-Pancreato-Biliary Surgery, Institute of Clinical Medicine, University of Oslo, Oslo, Norway.
⁶ Department of Hepato-Pancreato-Biliary Surgery, Oslo University Hospital Rikshospitalet, Oslo, Norway.
⁷ Department of Pathology, Institute of Clinical Medicine, University of Oslo, Oslo, Norway.

Abstract

Background: Profound resistance to chemotherapy remains a major challenge in achieving better clinical outcomes for patients with pancreatic ductal adenocarcinoma (PDAC). Recent studies indicate that gemcitabine (GEM) resistance is promoted both by pancreatic stellate cells (PSCs) and through increased glycolysis. However, it remains unknown whether PSCs affect GEM sensitivity via glycolytic regulation.

Methods: Human pancreatic cancer cell (PCC) lines (BxPC-3, Capan-2, HPAF-II, Mia PaCa-2, Panc-1, SW-1990) were exposed to three different PSC-conditioned media (PSC-CM; PSC-1, PSC-2, HPaSteC), following either pre-treatment with glycolysis inhibitor NV-5440 or transfection for transient silencing of key glycolytic regulators (LDHA and MCT4). Proliferation, glucose transport, extracellular lactate, and GEM sensitivity were assessed. Protein expression was determined by Western blot and immunostaining. Moreover, secreted proteins in PSC-CMs were profiled by mass spectrometry (MS).

Results: While exposure to PSC-CMs did not affect glucose transport in PCCs, it increased their lactate release and proliferation, and reduced the sensitivity for GEM. Both NV-5440 treatment and transient silencing of LDHA and MCT4 inhibited these PSC-induced changes in PCCs. MS analysis identified 688 unique proteins with differential expression, of which only 87 were common to the three PSC-CMs. Most PSC-secreted proteins were extracellular matrix-related, including SPARC, fibronectin, and collagens. Moreover, exposure to PSC-CMs increased the phosphorylation of ERK in PCCs, but the treatment of PCCs with the MEK/ERK inhibitor PD98059 resulted in a reduction of PSC-CM-induced glycolysis and improved GEM sensitivity.

Conclusions: The study findings suggest that PSC-secreted factors promote both glycolysis and GEM resistance in PCCs, and that glycolysis inhibition by NV-5440 and blocking of ERK phosphorylation by PD98059 protect PCCs from PSC-CM-induced loss of GEM sensitivity. Taken together, PSCs appear to promote GEM resistance in PDAC via glycolysis. Thus, targeting glycolysis may improve the effect of chemotherapy in PDAC.

Keywords: Gemcitabine sensitivity; Glycolysis; Pancreatic cancer; Pancreatic stellate cells.

Abstract

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