Analysis of lysyl oxidase as a marker for diagnosis of canine mammary tumors.
Animals
Biomarkers, Tumor
/ metabolism
Dogs
/ metabolism
Female
Gene Expression Regulation, Neoplastic
/ genetics
Mammary Glands, Animal
/ metabolism
Mammary Neoplasms, Animal
/ diagnosis
Prognosis
Protein-Lysine 6-Oxidase
/ analysis
RNA, Messenger
/ genetics
Real-Time Polymerase Chain Reaction
/ methods
Recombinant Proteins
/ genetics
Transcriptome
/ genetics
Biomarker
Canine mammary tumor
Cloning
ELISA
Lysyl oxidase
Real time-PCR
Journal
Molecular biology reports
ISSN: 1573-4978
Titre abrégé: Mol Biol Rep
Pays: Netherlands
ID NLM: 0403234
Informations de publication
Date de publication:
Oct 2019
Oct 2019
Historique:
received:
03
11
2018
accepted:
25
06
2019
pubmed:
3
7
2019
medline:
10
3
2020
entrez:
3
7
2019
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 31264163
doi: 10.1007/s11033-019-04941-1
pii: 10.1007/s11033-019-04941-1
doi:
Substances chimiques
Biomarkers, Tumor
0
RNA, Messenger
0
Recombinant Proteins
0
Protein-Lysine 6-Oxidase
EC 1.4.3.13
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
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