Pancreatic cancer (PC), a malignant neoplasm of the gastrointestinal tract, is linked to dismal prognoses and a restricted response to chemotherapy. Long noncoding RNAs (lncRNAs) play critical roles in regulating PC stemness. This study investigated the mechanism of cancer-associated fibroblasts (CAFs)-derived lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) promoting PC cell stemness via the microRNA-101-3p (miR-101-3p)/lysine demethylase 3A (KDM3A) axis. Human pancreatic CAFs were cultured in vitro and transfected with small interfering (si)-NEAT1. Extracellular vesicles (EVs) were isolated from CAFs using ultracentrifugation and identified using transmission electron microscopy, nanoparticle tracking analysis, and western blot. Human PC cells PANC-1 were treated with miR-101-3p mimic, si-KDM3A, and EVs. Expression of NEAT1, miR-101-3p, KDM3A, achaete-scute family BHLH transcription factor 2 (ASCL2), leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5), octamer-binding transcription factor 4 (Oct4), and SRY-box transcription factor 2 (SOX2), CD44+CD133+ cell count, and cell malignant biological behaviors were assessed. miR-101-3p-KDM3A targeted binding sites were predicted by StarBase database, with the targeted relationship verified by the dual-luciferase reporter assay. After CAFs-derived EVs (CAFs-EVs) treatment, PANC-1 cells exhibited increased NEAT1, ASCL2, LGR5, Oct4, and SOX2 expression, cell malignant biological behaviors and stemness ability, and CD44+CD133+ cell count. CAFs-EVs-delivered NEAT1 potentiated PANC-1 cell stemness by downregulating miR-101-3p. NEAT1 upregulated KDM3A expression by inhibiting miR-101-3p. KDM3A knockdown partly reversed CAFs-EVs-carried NEAT1-promoted PANC-1 cell stemness. CAFs-EVs-shuttled NEAT1 encouraged PANC-1 cell stemness via the miR-101-3p/KDM3A axis. CAFs-EVs might carry lncRNA NEAT1 to inhibit miR-101-3p expression, thereby regulating KDM3A and eventually facilitating PC cell stemness.
Li et al. (Wed,) studied this question.