Histological inspection of visually normal tissue adjacent to neoplastic lesions often reveals multiple foci of cellular abnormalities. This suggests the presence of a regional carcinogenic signal that spreads oncogenic transformation and field cancerization. We observed an abundance of mutagenic
reactive oxygen species in the stroma of cryosectioned patient
tumor biopsies, indicative of extratumoral oxidative stress. Diffusible
hydrogen peroxide (H2O2) was elevated in the
conditioned medium of cultured skin epithelia at various stages of oncogenic transformation, and H2O2 production increased with greater
tumor-forming and metastatic capacity of the studied cell lines. Explanted cancer-associated fibroblasts (CAFs) also had higher levels of H2O2 secretion compared with normal fibroblasts (
FIBs). These results suggest that extracellular H2O2 acts as a field effect
carcinogen. Indeed, H2O2-treated keratinocytes displayed decreased
phosphatase and
tensin homolog (PTEN) and increased Src activities because of oxidative modification. Furthermore, treating
FIBs with CAF-
conditioned medium or exogenous H2O2 resulted in the acquisition of an oxidative, CAF-like state. In vivo, the proliferative potential and invasiveness of composite
tumor xenografts comprising cancerous or non-
tumor-forming epithelia with CAFs and
FIBs could be attenuated by the presence of
catalase. Importantly, we showed that oxidatively transformed
FIBs isolated from composite
tumor xenografts retained their ability to promote
tumor growth and aggressiveness when adoptively transferred into new xenografts. Higher H2O2 production by CAFs was contingent on impaired TGFβ signaling leading to the suppression of the
antioxidant enzyme glutathione peroxidase 1 (GPX1). Finally, we detected a reduction in Smad3, TAK1 and TGFβRII expression in a cohort of 197 clinical
squamous cell carcinoma (SCC) CAFs, suggesting that impaired stromal TGFβ signaling may be a clinical feature of SCC. Our study indicated that CAFs and
cancer cells engage redox signaling circuitries and mitogenic signaling to reinforce their reciprocal relationship, suggesting that future anticancer approaches should simultaneously target
ligand receptor and redox-mediated pathways.