Celecoxib induces proliferation and Amphiregulin production in colon subepithelial myofibroblasts, activating erk1–2 signaling in synergy with EGFR

The COX-2 inhibitor Celecoxib, tested in phase III trials for the prevention of sporadic colon adenomas, reduced the appearance of new adenomas, but was unable to affect the incidence of colon cancer. Moreover the 5 years follow-up showed that patients discontinuing Celecoxib treatment had an increased incidence of adenomas as compared to the placebo arm. In the APC(min/+) mouse model short term treatment with Celecoxib reduced gut adenomas, but a prolonged administration of the drug induced fibroblast activation and intestinal fibrosis with a final tumor burden. The way Celecoxib could directly activate human colon myofibroblasts (MF) has not yet been investigated.We found that MF are activated by non toxic doses of Celecoxib. Celecoxib induces erk1–2 and Akt phosphorylation within 5′. This short term activation is apparently insufficient to cause phenotypic changes, but the contemporary triggering of EGFR causes an impressive synergic effect inducing MF proliferation and the neo-expression and release of Amphiregulin (AREG), a well known EGFR agonist involved in colon cancer progression. As a confirm to these observations, the erk inhibitor U0126 and the EGFR inhibitors Tyrphostin and Cetuximab were able to contrast AREG induction.Our data provide evidence that Celecoxib directly activates MF empowering EGFR signaling. According to these results the association with EGFR (or erk1–2) inhibitors could abolish the off-target activity of Celecoxib, possibly extending the potential of this drug for colon cancer prevention.

► We used primary cultures of colorectal myofibroblasts (MF) to investigate the effects of non-toxic doses of Celecoxib.
► Celecoxib induced MF proliferation and Amphiregulin secretion in cultures derived from either normal, or tumoral mucosa.
► Celecoxib activity was mediated by the rapid phosphorylation of erk1-2 and Akt.
► Only erk1-2 signaling was involved in Amphiregulin secretion.
► The triggering of EGFR caused an exponential increase of Celecoxib activity.