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. 2016 Jun 7;7:144–149. doi: 10.1016/j.bbrep.2016.06.004

Fig. 1.

Fig. 1

Adipocytes promote PanIN and PDAC cell proliferation in a glutamine-dependent manner: (A) 3T3L1 adipocytes: Top: Photomicrograph of Oil Red-O-stained mature 3T3L1 adipocytes on standard plastic (left) and on a transwell insert (right). Bottom: Upregulation of adipogenic gene transcripts (PPAR-γ, FASN, ATGL, SREBP1c, CEBP-α) based on QRTPCR in mature 3T3L1 adipocytes relative to undifferentiated 3T3L1 preadipocytes cultured on standard plastic or on a transwell insert. Ordinate: fold increase in transcript levels in mature adipocytes relative to transcript levels in undifferentiated 3T3L1 cells as a referent=1; *: p<0.001 comparing transcript levels in mature adipocytes to undifferentiated 3T3L1 cell referent; n=5 experiments. (B) PanIN/PDAC cell proliferation is glutamine-dependent: PanIN/PDAC cell proliferation in monoculture for 72 h in low serum (0.5%FCS) glutamine-free media with the indicated glucose concentration (left), or in glucose-free media with the indicated glutamine (Gln) concentration (right). Ordinates: percent change in XTT signal relative to PanIN/PDAC cells in monoculture in nutrient-poor (glucose-free, glutamine-free) media as a referent set at zero; *: p<0.050 comparing experimental condition to referent PanIN/PDAC cell proliferation n=6 experiments. (C) Positive PanIN/PDAC cell proliferation trajectories are maintained in nutrient-poor media: PanIN/PDAC cell proliferation in monoculture for 72 h in low serum media with the indicated glucose and glutamine concentrations. Ordinates: percent change in XTT signal relative to PanIN/PDAC cells in monoculture at time-zero in the corresponding culture conditions as a referent=zero; *: p<0.050 comparing experimental condition to referent time-zero PanIN/PDAC cell proliferation n=7 experiments. (D) Adipocytes promote PDAC cell proliferation: PanIN/PDAC cell proliferation after 72 h in low serum media with the indicated glucose and glutamine concentrations with one of the following in an overlying transwell insert: mature differentiated adipocytes (Ad CC); mature adipocyte-conditioned media (Ad CM); or undifferentiated pre-adipocytes (Undiff CC). Ordinates: percent increase in XTT signal relative to PanIN/PDAC cells in monoculture in the same substrate conditions indicated in each graph as a referent=zero; *: p<0.050 comparing experimental condition to referent PanIN/PDAC cell proliferation n=6 experiments. (E) Adipocytes rescue PDAC cell proliferation in a glutamine-dependent manner: PanIN/PDAC cell proliferation after 72 h culture in low serum glucose-free media and (left to right): monoculture with 4 mM glutamine (MC, Gln 4 mM), co-culture with mature adipocytes in 0 mM glutamine (Ad CC, Gln 0 mM), co-culture with mature adipocytes pre-treated for 24 h with l-methionine sulfoximine in 0 mM glutamine (Ad CC/GSI, Gln 0 mM), co-culture with adipocytes and GPNA with 0 mM glutamine (Ad CC, GPNA, Gln 0 mM), or monoculture with GPNA in 4 mM glutamine (MC, GPNA, Gln 4 mM). Ordinate: percent change in XTT signal in experimental arm relative to PanIN/PDAC cells in monoculture in low serum glucose-free, glutamine-free media as a referent=0; *: p<0.050 comparing experimental condition to referent PanIN/PDAC cell proliferation n=7 experiments.