OBJECTIVE We previously showed that human pancreatic stellate cells(HPSCs)promote pancreatic ductal adenocarcinoma(PDAC)cell growth by activating nuclear factor erythroid2-related factor 2(Nrf2),a key transcriptional regulator of cytoprotective genes.We aim to investigate whether Nrf2-mediated metabolic reprogramming and reactive oxygen species(ROS)detoxification are involved in HPSCs-mediated cell growth.METHODS Nrf2-mediated metabolic genes expression of pentose phosphate pathway(PPP)for purine nucleotide synthesis;glutamine metabolism for nicotinamide adenine dinucleotide phosphate(NADPH)-equivalent producers and also glutathione biosynthesis both for intracellular ROS inactivation were examined using quantitative real-time PCR(qRT-PCR)after treated with conditioned media derived from HPSCs(HPSC-CM)in human PDAC cells(BxPC-3and AsPC-1)with or without Nrf2 gene silencing using siRNA-mediated technique.Metabolites involved in PPP for purine nucleotide and NADPH generation were selected and their concentration was measured using UHPLC-MS/MS.Antioxidants,tiron and N-acetylcysteine(NAC)were used to attenuate the intracellular ROS rendered by Nrf2 before measuring PDAC cell growth and also phosphorylation of extracellular signal-regulated kinase(ERK)1/2and protein kinase B(AKT)using MTT and Western blotting,respectively.RESULTS Metabolically,HPSC-CMupregulated Nrf2-mediated genes involved in three metabolic pathways(G6PD,PGD,TKT,PPAT,MTHFD2,ME1,IDH1,GCLC and GCLM)in BxPC-3and AsPC-1cells.HPSC-CM was able to upregulate all the metabolic genes after Nrf2 gene silencing,and also significantly increased the metabolite concentration of ribose 5-phosphate and inosine 5′-monophosphate,which are involved in nucleotide synthesis for cell growth.Decreasing the intracellular ROS rendered by Nrf2 suppressed PDAC cell growth and also phosphorylation of ERK 1/2and AKT protein.CONCLUSION Our findings reveal that HPSC-CM activates Nrf2-mediated metabolic reprogramming,which leads to purine nucleotide synthesis and ROS detoxification to promote PDAC cell growth. OBJECTIVE We previously showed that human pancreatic stellate cells (HPSCs) promote pancreatic ductal adenocarcinoma (PDAC) cell growth by activating nuclear factor erythroid 2-related factor 2 (Nrf2), a key transcriptional regulator of cytoprotective genes. We aim to investigate whether Nrf2-mediated metabolic reprogramming and reactive oxygen species (ROS) detoxification are involved in HPSCs-mediated cell growth. METHODS Nrf2 -mediated metabolic genes expression of pentose phosphate pathway (PPP) for purine nucleotide synthesis; glutamine metabolism for nicotinamide adenine dinucleotide phosphate (NADPH) -equivalent producers and also glutathione biosynthesis both for intracellular ROS inactivation were examined using quantitative real-time PCR (qRT-PCR) after treated with conditioned media derived from HPSCs (HPSC-CM) in human PDAC cells (BxPC-3 and AsPC-1) with or without Nrf2 gene silencing using siRNA-mediated technique. Metabolites involved in PPP for purine nucleotide and NADPH generation were selecte d and their concentration was measured using UHPLC-MS / MS. Antioxidants, tiron and N-acetylcysteine ​​(NAC) were used to attenuate the intracellular ROS rendered by Nrf2 before measuring PDAC cell growth and also phosphorylation of extracellular signal- regulated kinase (ERK) 1 / 2and protein kinase B (AKT) using MTT and Western blotting, respectively.RESULTS Metabolically, HPSC-CMupregulated Nrf2-mediated genes involved in three metabolic pathways (G6PD, PGD, TKT, PPAT, MTHFD2, ME1, IDH1, GCLC and GCLM ) in BxPC-3 and AsPC-1 cells. HPC-CM was able to upregulate all the metabolic genes after Nrf2 gene silencing, and also significantly increased the metabolite concentration of ribose 5-phosphate and inosine 5’-monophosphate, which are involved in nucleotide synthesis for cell growth. Act for rendering the intracellular ROS rendered by Nrf2 suppressed PDAC cell growth and also phosphorylation of ERK 1/2 and AKT protein. CONCLUSION Our findings reveal that HPSC-CM activates Nrf2-mediated metabolic reprogramming, which leads to purine nucleotide synthesis and ROS detoxification to promote PDAC cell growth.