Fig. 3.

Acetylcholinesterase (AChE) inhibition attenuates the formation of xenografted PCa in mice and decreases tumor-associated inflammation. a In vivo xenograft model: human T3M-4 pancreatic cancer cells were transplanted subcutaneously into Crl:NMRI-^Foxn1nu/nu mice that were treated prophylactically or therapeutically, the latter beginning at 1 week after tumor induction. Physostigmine and pyridostigmine were administered daily s.c. at 0.1 x LD₅₀ (low dose) or 0.3 x LD₅₀ (high dose), and Pyridostigmine at 0.2 x LD₅₀ (low dose) and 0.4 x LD₅₀ (low dose)or saline injection (control). b Prophylactic treatment with physostigmine or pyridostigmine resulted in a reduction of the xenografted tumor mass (means ± SD). c Therapeutic treatment with AChE inhibitors in mice xenografted with T3M-4 cells. d show the percentage of animals that had invasive penetrating tumor growth (Control animals 8/10 = 80% and prophylactically treated animals 6/40 = 15%) e Representative IF photomicrograph of double-positive tumor-associated macrophages (TAMs) stained with CD45, f4/80 and DAPI. Graph shows the amount of double-positive cells per square centimeters of tumor tissue compared to the untreated animals (****p < 0.0001 for Phys. low-prophyl., Phys. high-prophyl., and Pyrido. high prophyl. Respectively; Pyrido. low-prophyl. *p = 0.0370 by unpaired t-test). Means ± SEM. * p < 0.05; *** p < 0.001; **** p < 0.0001). f Comparative analysis of cytokine levels in the serum of physostigmine or pyridostigmine treated, xenografted mice (“treated”) in comparison with control/saline treated mice (“control”). The results of prophylactically and therapeutically treated animals have been pooled in the graphs