title: Selective depletion of tumour suppressors Deleted in Colorectal Cancer (DCC) and neogenin by environmental and endogenous serine proteases: linking diet and cancer.
abstract: The related tumour suppressor proteins Deleted in Colorectal Cancer (DCC) and neogenin are absent or weakly expressed in many cancers, whereas their insertion into cells suppresses oncogenic behaviour. Serine proteases influence the initiation and progression of cancers although the mechanisms are unknown.
  
  
  
  The effects of environmental (bacterial subtilisin) and endogenous mammalian (chymotrypsin) serine proteases were examined on protein expression in fresh, normal tissue and human neuroblastoma and mammary adenocarcinoma lines. Cell proliferation and migration assays (chemoattraction and wound closure) were used to examine cell function. Cells lacking DCC were transfected with an ectopic dcc plasmid.
  
  
  
  Subtilisin and chymotrypsin selectively depleted DCC and neogenin from cells at nanomolar concentrations without affecting related proteins. Cells showed reduced adherence and increased migration, but after washing they re-attached within 24 h, with recovery of protein expression. These effects are induced by chymotryptic activity as they are prevented by chymostatin and the soybean Bowman-Birk inhibitor typical of many plant protease inhibitors.
  
  
  
  Bacillus subtilis, which secretes subtilisin is widely present in soil, the environment and the intestinal contents, while subtilisin itself is used in meat processing, animal feed probiotics and many household cleaning agents. With chymotrypsin present in chyme, blood and tissues, these proteases may contribute to cancer development by depleting DCC and neogenin. Blocking their activity by Bowman-Birk inhibitors may explain the protective effects of a plant diet. Our findings identify a potential non-genetic contribution to cancer cell behaviour which may explain both the association of processed meats and other factors with cancer incidence and the protection afforded by plant-rich diets, with significant implications for cancer prevention.
authors.affiliation: College of Medical, Veterinary and Life Sciences, West Medical Building, University of Glasgow, Glasgow, G12 8QQ, UK.
authors.forename: Caroline M
authors.initials: CM
authors.lastname: Forrest
authors.affiliation: College of Medical, Veterinary and Life Sciences, West Medical Building, University of Glasgow, Glasgow, G12 8QQ, UK.
authors.forename: Kara
authors.initials: K
authors.lastname: McNair
authors.affiliation: College of Medical, Veterinary and Life Sciences, West Medical Building, University of Glasgow, Glasgow, G12 8QQ, UK.
authors.forename: Maria C J
authors.initials: MC
authors.lastname: Vincenten
authors.affiliation: Internal Medicine, Ashtead Hospital, Ashtead, Surrey, KT21 2SB, UK.
authors.forename: L Gail
authors.initials: LG
authors.lastname: Darlington
authors.affiliation: College of Medical, Veterinary and Life Sciences, West Medical Building, University of Glasgow, Glasgow, G12 8QQ, UK. Trevor.Stone@glasgow.ac.uk.
authors.forename: Trevor W
authors.initials: TW
authors.lastname: Stone
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement; D015179:Colorectal Neoplasms / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D000075389:DCC Receptor; D004032:Diet; D004783:Environmental Microbiology; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D059748:Proteolysis; D017208:Rats, Wistar; D011956:Receptors, Cell Surface / Q000378:metabolism*; D057057:Serine Proteases / Q000502:physiology*; D025521:Tumor Suppressor Proteins / Q000378:metabolism*
chemical_list: D000075389:DCC Receptor; C501909:DCC protein, human; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; D025521:Tumor Suppressor Proteins; C091279:neogenin; D057057:Serine Proteases
keywords: Chymotrypsin; DCC; Deleted in colorectal cancer; Neogenin; Serine proteases; Subtilisin