Analysis of lysyl oxidase as a marker for diagnosis of canine mammary tumors
Autor: | B. V. Sunil Kumar, Satparkash Singh, Afnan Saleem, Jaspreet Singh Arora, Ratan K. Choudhary |
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Rok vydání: | 2019 |
Předmět: |
0301 basic medicine
Sequence analysis Mammary Neoplasms Animal Lysyl oxidase Molecular cloning Biology Real-Time Polymerase Chain Reaction Protein-Lysine 6-Oxidase 03 medical and health sciences Dogs Mammary Glands Animal 0302 clinical medicine Gene expression Biomarkers Tumor Genetics Animals RNA Messenger Molecular Biology Gene Mammary tumor integumentary system General Medicine Prognosis Molecular biology Recombinant Proteins Gene Expression Regulation Neoplastic 030104 developmental biology Real-time polymerase chain reaction 030220 oncology & carcinogenesis biology.protein Female Transcriptome Elastin |
Zdroj: | Molecular Biology Reports. 46:4909-4919 |
ISSN: | 1573-4978 0301-4851 |
Popis: | Lysyl oxidase (LOX) is an extracellular metalloenzyme which mediates crosslinking of collagen and elastin. It has been reported to play a pivotal role in cancer metastasis especially in women suffering from breast cancer. The present study is the first to evaluate the gene expression levels of LOX by Real time-polymerase chain reaction (Real time-PCR) in dogs with mammary tumor besides molecular cloning and expression of canine lysyl oxidase gene (lox). Real time-PCR studies showed a significant upregulation (threefold higher) of lox in mammary tumor cases as compared to healthy dogs indicating its possible diagnostic and prognostic role in canine mammary tumors (CMTs). Cloning and sequencing of lox gene revealed 1230 bp CDS which is mostly conserved in C-terminal region. Sequence analysis of canine lox showed that it shares 99% homology with the predicted sequence available on NCBI and had greatest identity with the lox gene from cat. Protein structure predicted with homology modelling was validated by Ramachandran plot analysis which revealed most (approximately 95%) of the amino acids in favoured region. Additionally, recombinant lysyl oxidase expressed as His-tagged fusion protein in prokaryotic expression vector (pPROExHTa) was used in an ELISA for detection of circulating protein LOX in serum of CMT subjects. Receiver operating characteristics analysis of the ELISA revealed high sensitivity (90%) and specificity (85%) with histopathology as reference standard. Taken together, we propose LOX as a diagnostic biomarker and a putative prognostic candidate in CMT cases. |
Databáze: | OpenAIRE |
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