Cancer‐associated fibroblasts promote stemness maintenance and gemcitabine resistance via HIF‐1α/miR‐21 axis under hypoxic conditions in pancreatic cancer

Abstract

Gemcitabine (GEM) resistance affects chemotherapy efficacy of pancreatic cancer (PC). Cancer‐associated fibroblasts (CAFs) possess the ability of regulating chemoresistance. This study probed the mechanism of hypoxia‐treated CAFs regulating cell stemness and GEM resistance in PC. Miapaca‐2/SW1990 were co‐cultured with PC‐derived CAFs under normoxic/hypoxic conditions. Cell viability/self‐renewal ability was determined by MTT/sphere formation assays, respectively. Protein levels of CD44, CD133, Oct4, and Sox2 were determined by western blot. GEM tumoricidal assay was performed. PC cell GEM resistance was evaluated by MTT assay. CAFs were cultured at normoxia/hypoxia. HIF‐1α and miR‐21 expression levels were assessed by RT‐qPCR and western blot, with their binding sites and binding relationship predicted and verified. CAF‐extracellular vesicles (EVs) were incubated with Miapaca‐2 cells. The RAS/AKT/ERK pathway activation was detected by western blot. PC xenograft models were established and treated with hypoxic CAF‐EVs and GEM. CAFs and PC cell co‐culture increased cell stemness maintenance, GEM resistance, cell viability, stem cell sphere number, and protein levels of CD44, CD133, Oct4, and Sox2, and weakened GEM tumoricidal ability to PC cells, with the effects further enhanced by hypoxia. Hypoxia induced HIF‐1α and miR‐21 overexpression in CAFs. Hypoxia promoted CAFs to secrete high‐level miR‐21 EVs via the HIF‐1α/miR‐21 axis, and activated the miR‐21/RAS/AKT/ERK pathway. CAF‐EVs promoted GEM resistance in PC via the miR‐21/RAS/ATK/ERK pathway in vivo. Hypoxia promoted CAFs to secrete high‐level miR‐21 EVs through the HIF‐1α/miR‐21 axis, and activated the miR‐21/RAS/AKT/ERK pathway via EVs to trigger stemness maintenance and GEM resistance in PC.

Hypoxia-Inducible Factor-Dependent and Independent Mechanisms Underlying Chemoresistance of Hypoxic Cancer Cells

“…This circRNA sponges miR-23b-3p to enhance SIRT1-HIF-1α signaling in recipient cells and thereby induces gemcitabine resistance in cell culture and xenografted tumor models [110]. In a separate study, it has been shown that HIF-induced exosomal miR-21 from hypoxic CAFs activates the RAS-AKT-ERK pathway and promotes gemcitabine resistance in the recipient pancreatic cancer cells both in vitro and in vivo [111].…”