79223ad80f59973603171d3fda88baa1 IJMS_a2024v25p1170.pdf 007e265b58f08903b24abf0a9f20c8783344730d IJMS_a2024v25p1170.pdf 439055abd453dddcbbc107adc9d70f907bb2903574100af6365c174c6957dce2 IJMS_a2024v25p1170.pdf Title: Programmed Death-Ligand (PD-L1), Epidermal Growth Factor (EGF), Relaxin, and Matrix Metalloproteinase-3 (MMP3): Potential Biomarkers of Malignancy in Canine Mammary Neoplasia Subject: Gene expression has been suggested as a putative tool for prognosis and diagnosis in canine mammary neoplasia (CMNs). In the present study, 58 formalin-fixed paraffin-embedded (FFPE) paraffined canine mammary neoplasias from 27 different bitches were included. Thirty-seven tumours were classified as benign, whereas thirty-one were classified as different types of canine carcinoma. In addition, mammary samples from three healthy bitches were also included. The gene expression for vascular endothelial growth factor- (VEGF), CD20, progesterone receptor (PGR), hyaluronidase-1 (HYAL-1), programmed death-ligand 1 (PD-L1), epidermal growth factor (EGF), relaxin (RLN2), and matrix metalloproteinase-3 (MMP3) was assessed through RT-qPCR. All the assessed genes yielded a higher expression in neoplastic mammary tissue than in healthy tissue. All the evaluated genes were overexpressed in neoplastic mammary tissue, suggesting a role in the process of tumorigenesis. Moreover, PD-L1, EGF, relaxin, and MMP3 were significantly overexpressed in malignant CMNs compared to benign CMNs, suggesting they may be useful as malignancy biomarkers. Keywords: canine mammary neoplasia; gene expression; malignancy biomarkers; programmed death-ligand 1; epidermal growth factor; relaxin; matrix metalloprotease-3 Author: Makchit Galadima, Mariana Teles, Josep Pastor, Javier Hernández-Losa, Joan Enric Rodríguez-Gil and Maria Montserrat Rivera del Alamo Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Thu Jan 18 08:50:52 2024 CET ModDate: Thu Jan 18 08:58:38 2024 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 15 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1250297 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- SSNIUI+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 ZXDKIP+EURM10 Type 1 Builtin yes yes yes 16 0 ZGBBXZ+URWPalladioL-Bold Type 1 Custom yes yes yes 21 0 LUUYRB+URWPalladioL-Roma Type 1 Custom yes yes yes 27 0 WLWMDZ+URWPalladioL-Ital Type 1 Custom yes yes yes 32 0 IOLAWT+PazoMath-Italic Type 1 Builtin yes yes yes 37 0 GFKJOY+VnURWPalladioL Type 1 Custom yes yes yes 42 0 IRWHZX+TimesNewRomanPSMT Type 1C WinAnsi yes yes no 58 0 CKUPDE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 61 0 AAPCWM+CMSY10 Type 1 Builtin yes yes yes 81 0 FPGWUO+PalatinoLinotype,Italic CID TrueType Identity-H yes yes yes 91 0 FPTDMR+PalatinoLinotype,BoldItalic CID TrueType Identity-H yes yes yes 97 0 FPGWUO+PalatinoLinotype,Bold CID TrueType Identity-H yes yes yes 103 0 FPJGZF+PalatinoLinotype CID TrueType Identity-H yes yes yes 109 0 KZCBCP+URWPalladioL-BoldItal Type 1 Custom yes yes yes 148 0 LELDCK+PazoMath-BoldItalic Type 1 Builtin yes yes yes 153 0 PAOLAL+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 170 0 PAOLAM+PalatinoLinotype CID TrueType Identity-H yes yes yes 173 0 PAOLBM+PalatinoLinotype TrueType WinAnsi yes yes no 179 0 PAOLBO+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 182 0 PAOLGN+PalatinoLinotype TrueType MacRoman yes yes no 185 0 MPUDFL+PazoMath Type 1 Builtin yes yes yes 204 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-02-06 02:30:34 CET RepresentationInformation: IJMS_a2024v25p1170.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-02-05 12:25:57 CET Size: 1250297 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 458 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results Destination: section.2 Children: Item: Title: Tumours Typification Destination: subsection.2.1 Item: Title: Estimation of Gene Efficiency Destination: subsection.2.2 Item: Title: Correlation between Genes Destination: subsection.2.3 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Animals and Sampling Destination: subsection.4.1 Item: Title: Extraction of RNA and Synthesis of Complementary DNA Destination: subsection.4.2 Item: Title: Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR) Destination: subsection.4.3 Item: Title: Standardization Strategy Destination: subsection.4.4 Item: Title: Statistical Analysis Destination: subsection.4.5 Item: Title: Conclusions Destination: section.5 Item: Title: Appendix A Destination: appendix.A. Item: Title: References Destination: appendix.B. Info: Title: Programmed Death-Ligand (PD-L1), Epidermal Growth Factor (EGF), Relaxin, and Matrix Metalloproteinase-3 (MMP3): Potential Biomarkers of Malignancy in Canine Mammary Neoplasia Author: Makchit Galadima, Mariana Teles, Josep Pastor, Javier Hernández-Losa, Joan Enric Rodríguez-Gil and Maria Montserrat Rivera del Alamo Subject: Gene expression has been suggested as a putative tool for prognosis and diagnosis in canine mammary neoplasia (CMNs). In the present study, 58 formalin-fixed paraffin-embedded (FFPE) paraffined canine mammary neoplasias from 27 different bitches were included. Thirty-seven tumours were classified as benign, whereas thirty-one were classified as different types of canine carcinoma. In addition, mammary samples from three healthy bitches were also included. The gene expression for vascular endothelial growth factor- (VEGF), CD20, progesterone receptor (PGR), hyaluronidase-1 (HYAL-1), programmed death-ligand 1 (PD-L1), epidermal growth factor (EGF), relaxin (RLN2), and matrix metalloproteinase-3 (MMP3) was assessed through RT-qPCR. All the assessed genes yielded a higher expression in neoplastic mammary tissue than in healthy tissue. All the evaluated genes were overexpressed in neoplastic mammary tissue, suggesting a role in the process of tumorigenesis. Moreover, PD-L1, EGF, relaxin, and MMP3 were significantly overexpressed in malignant CMNs compared to benign CMNs, suggesting they may be useful as malignancy biomarkers. 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