title: Expression and characterization of tissue inhibitor of metalloproteinase 4 from complex canine mammary carcinomas.
abstract: All four members of the tissue inhibitors of metalloproteinase (TIMP) family have been reported to be over-expressed in breast cancer cells in vitro. Dysregulation of TIMP-4 expression predicts poor prognosis in cancers. The present study evaluated the association of the expression levels of TIMP-4 with mammary tumor development in dogs, measured by real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). Mammary tissue samples were collected from healthy canine mammary gland and from tumor subjects. TIMP-4 expression was found to be upregulated (5.856 times) in complex canine mammary carcinomas. Also, TIMP-4 mature peptide was expressed heterologously in E. coli. The recombinant protein was purified by Ni- NTA affinity chromatography and further confirmed by western blotting. The rTIMP-4 was found to be functionally active and could inhibit matrix metalloproteinase 11(MMP-11) activity. Immunization of mice with rTIMP-4 resulted in increased antigen specific serum antibody titer, and this serum could be suitably used to detect and quantify the protein in sera of dogs with mammary tumors. TIMP-4 could act as a marker of canine mammary tumors. To the best of our knowledge, this is the first report of heterologous expression of TIMP-4 from complex canine mammary carcinomas.
authors.affiliation: a School of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University , Ludhiana , India.
authors.forename: Navneet
authors.initials: N
authors.lastname: Kaur
authors.affiliation: a School of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University , Ludhiana , India.
authors.forename: B V
authors.initials: BV
authors.lastname: Sunil Kumar
authors.affiliation: a School of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University , Ludhiana , India.
authors.forename: Kanika
authors.initials: K
authors.lastname: Mahajan
authors.affiliation: a School of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University , Ludhiana , India.
authors.forename: Satparkash
authors.initials: S
authors.lastname: Singh
mesh_terms: D000818:Animals; D004283:Dog Diseases / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D004285:Dogs; D005260:Female; D006801:Humans; D008297:Male; D015674:Mammary Neoplasms, Animal / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D060888:Real-Time Polymerase Chain Reaction; D019714:Tissue Inhibitor of Metalloproteinases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D000097792:Tissue Inhibitor of Metalloproteinase-4
chemical_list: D019714:Tissue Inhibitor of Metalloproteinases; D000097792:Tissue Inhibitor of Metalloproteinase-4
keywords: ELISA; TIMP-4; canine; expression profiling; mammary tumor