Figure 1.

PAMAM-G3 Inhibits TLR-9-Activating, Pro-invasive DAMPs in Pancreatic Cancer

(A) Serum cfDNA levels in healthy individuals, PC patients with localized, early-stage disease before and after CRT, and PC patients with known metastatic disease (n = 8 for all groups). (B) Activation of TLR-9-specific reporter cells by either healthy human sera or PC patient sera in the absence or presence of PAMAM-G3 (20 μg/mL). (C) Invasion of Panc1 PC cells in a transwell-Matrigel assay after addition of either healthy human sera or PC patient sera in the absence or presence of PAMAM-G3 (20 μg/mL). (D) Invasion of Panc1 cells after treatment with vehicle (media) or the TLR-9-specific agonist CpG ODN 2006 (5 μM) in the absence or presence of PAMAM-G3 (20 μg/mL). Effect of PAMAM-G3 alone on Panc1 cell invasion is also shown. (E) Cell viability as measured by Cell-titer Glo luminescence assay was determined after incubation of Panc1 PC cells with vehicle (media), CpG ODN (5 μM), PAMAM-G3 (20 μg/mL), or 1% Triton X-100 for 24 hr. (F) Invasion of KPC4580P PC cells in a transwell-Matrigel assay after addition of either healthy human sera, PC patient sera, or PC patient sera in the presence of PAMAM-G3 (20 μg/mL) or the TLR 9 inhibitor ODN 2088. (G) Effect of vehicle (media) or CpG ODN 2006 (5 μM) treatment, alone or in combination with PAMAM-G3 (20 μg/mL), on nuclear translocation of NF-κB in BxPC3 PC cells. Bar graphs denote mean ± SEM. TLR 9 activation, PC cell invasion and viability, and NF-κB translocation experiments were repeated at least three times, and figures depict a single representative experiment. HPF, high powered field; RLU, relative light units. ****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05; and NS, not significant, by two-tailed t test.