Expression and characterization of tissue inhibitor of metalloproteinase 4 from complex canine mammary carcinomas. Issue 5 (2nd September 2016)

• Record Type: Journal Article
• Authors: Kaur, Navneet; Sunil Kumar, B. V.; Mahajan, Kanika; Singh, Satparkash
• Is Part Of: Journal of immunoassay & immunochemistry. Volume 37:Issue 5(2016)
• Page Start: 515
• Page End: 526
• Publication Date: 2016-09-02
• Subjects: mammary tumor -- canine -- TIMP-4 -- expression profiling -- ELISA; Immunoassay -- Periodicals; Immunochemistry -- Periodicals; Immunoassay -- periodicals; Immunochemistry -- periodicals; 571.96405
• DOI: 10.1080/15321819.2016.1162800
• Languages: English
• ISSNs: 1532-1819

Abstract

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.