Immunohistochemical Analysis of Inter-Alpha-Trypsin Inhibitor Heavy Chain 2 and Enolase 1 in Canine Mammary Tumors: Associations with Tumor Aggressiveness and Prognostic Significance
Mammary neoplasms in dogs are a common clinical concern, especially in middle-aged and older intact females. These tumors share similarities with human breast cancer in terms of histology, disease progression, and risk factors, making dogs a relevant model for breast cancer research. The search for...
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2025-02-01
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| author | Luadna dos Santos e Silva Pedro Henrique Fogaça Jordão Beatriz Castilho Balieiro Laura de Souza Baracioli Daniela Farias de Nóbrega Adriana Alonso Novais Luiz Gustavo de Almeida Chuffa Debora Aparecida Pires de Campos Zuccari |
| author_facet | Luadna dos Santos e Silva Pedro Henrique Fogaça Jordão Beatriz Castilho Balieiro Laura de Souza Baracioli Daniela Farias de Nóbrega Adriana Alonso Novais Luiz Gustavo de Almeida Chuffa Debora Aparecida Pires de Campos Zuccari |
| author_sort | Luadna dos Santos e Silva |
| collection | DOAJ |
| description | Mammary neoplasms in dogs are a common clinical concern, especially in middle-aged and older intact females. These tumors share similarities with human breast cancer in terms of histology, disease progression, and risk factors, making dogs a relevant model for breast cancer research. The search for biomarkers in canine mammary tumors is essential to understand tumor progression and identify potential therapeutic targets. This study investigated the expression of two potential biomarkers—Inter-Alpha-Trypsin Inhibitor Heavy Chain 2 (ITIH2) and Enolase 1 (ENO1)—in the mammary glands of healthy and tumor-bearing dogs using immunohistochemistry. Both proteins were identified in previous proteomic analyses of extracellular vesicles derived from the plasma of healthy and tumor-bearing dogs. A total of fifty-one canine mammary tissue samples were analyzed and categorized into three groups: (i) the control group, composed of five samples of normal mammary tissue without neoplasia; (ii) benign tumors, composed of nineteen samples of benign mixed tumors; and (iii) malignant tumors, which included six carcinomas in grade 1 mixed tumors, five carcinomas in grade 2 mixed tumors, thirteen solid carcinomas of grade 3, one papillary carcinoma, and two tubular carcinomas. Regarding the intensity of staining, quantified by histoscore, there were no significant differences in the comparison between the groups; for ITIH2, the <i>p</i>-value was 0.33, and for ENO1, the <i>p</i>-value was 0.57. Regarding the predictive potential of their respective ROC curves, the proteins demonstrated low predictive power in canine mammary tumors. These findings indicate that neither ITIH2 nor ENO1 demonstrated strong prognostic value in this setting, as demonstrated by their moderate AUC values, wide confidence intervals, and lack of statistical significance. However, this study found distinct tissue localization patterns for ITIH2 and subcellular localization for ENO1. As an additional way to examine possible associations of these proteins with epithelial–mesenchymal transition, the ZEB1 antibody was tested by both single and double immunohistochemistry, demonstrating a tendency to be more intensely expressed in the malignant group and tending to be associated with ENO1 in canine mammary tumors. |
| format | Article |
| id | doaj-art-1a1ea8d83ebe4bbd8538b94ae297b500 |
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| language | English |
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| spelling | doaj-art-1a1ea8d83ebe4bbd8538b94ae297b5002025-08-20T02:03:31ZengMDPI AGVeterinary Sciences2306-73812025-02-0112211010.3390/vetsci12020110Immunohistochemical Analysis of Inter-Alpha-Trypsin Inhibitor Heavy Chain 2 and Enolase 1 in Canine Mammary Tumors: Associations with Tumor Aggressiveness and Prognostic SignificanceLuadna dos Santos e Silva0Pedro Henrique Fogaça Jordão1Beatriz Castilho Balieiro2Laura de Souza Baracioli3Daniela Farias de Nóbrega4Adriana Alonso Novais5Luiz Gustavo de Almeida Chuffa6Debora Aparecida Pires de Campos Zuccari7Molecular Investigation of Cancer Laboratory (MICL), Department of Molecular Biology, Faculty of Medicine of São José do Rio Preto (FAMERP), São José do Rio Preto 15090-000, BrazilBiochemistry and Molecular Biology Research Center (NPBIM), Faculty of Medicine of São José do Rio Preto (FAMERP), São José do Rio Preto 15090-000, BrazilMolecular Investigation of Cancer Laboratory (MICL), Department of Molecular Biology, Faculty of Medicine of São José do Rio Preto (FAMERP), São José do Rio Preto 15090-000, BrazilMolecular Investigation of Cancer Laboratory (MICL), Department of Molecular Biology, Faculty of Medicine of São José do Rio Preto (FAMERP), São José do Rio Preto 15090-000, BrazilPat Animal Laboratory, São José do Rio Preto 15070-000, BrazilInstitute of Health Science (ICS), Universidade Federal de Mato Grosso (UFMT), Sinop 78550-728, BrazilDepartment of Structural and Functional Biology, Institute of Biosciences, UNESP, São Paulo State University, Botucatu 18618-689, BrazilMolecular Investigation of Cancer Laboratory (MICL), Department of Molecular Biology, Faculty of Medicine of São José do Rio Preto (FAMERP), São José do Rio Preto 15090-000, BrazilMammary neoplasms in dogs are a common clinical concern, especially in middle-aged and older intact females. These tumors share similarities with human breast cancer in terms of histology, disease progression, and risk factors, making dogs a relevant model for breast cancer research. The search for biomarkers in canine mammary tumors is essential to understand tumor progression and identify potential therapeutic targets. This study investigated the expression of two potential biomarkers—Inter-Alpha-Trypsin Inhibitor Heavy Chain 2 (ITIH2) and Enolase 1 (ENO1)—in the mammary glands of healthy and tumor-bearing dogs using immunohistochemistry. Both proteins were identified in previous proteomic analyses of extracellular vesicles derived from the plasma of healthy and tumor-bearing dogs. A total of fifty-one canine mammary tissue samples were analyzed and categorized into three groups: (i) the control group, composed of five samples of normal mammary tissue without neoplasia; (ii) benign tumors, composed of nineteen samples of benign mixed tumors; and (iii) malignant tumors, which included six carcinomas in grade 1 mixed tumors, five carcinomas in grade 2 mixed tumors, thirteen solid carcinomas of grade 3, one papillary carcinoma, and two tubular carcinomas. Regarding the intensity of staining, quantified by histoscore, there were no significant differences in the comparison between the groups; for ITIH2, the <i>p</i>-value was 0.33, and for ENO1, the <i>p</i>-value was 0.57. Regarding the predictive potential of their respective ROC curves, the proteins demonstrated low predictive power in canine mammary tumors. These findings indicate that neither ITIH2 nor ENO1 demonstrated strong prognostic value in this setting, as demonstrated by their moderate AUC values, wide confidence intervals, and lack of statistical significance. However, this study found distinct tissue localization patterns for ITIH2 and subcellular localization for ENO1. As an additional way to examine possible associations of these proteins with epithelial–mesenchymal transition, the ZEB1 antibody was tested by both single and double immunohistochemistry, demonstrating a tendency to be more intensely expressed in the malignant group and tending to be associated with ENO1 in canine mammary tumors.https://www.mdpi.com/2306-7381/12/2/110bitchesmammary tumorsextracellular vesicles (EVs)biomarkersprognosis |
| spellingShingle | Luadna dos Santos e Silva Pedro Henrique Fogaça Jordão Beatriz Castilho Balieiro Laura de Souza Baracioli Daniela Farias de Nóbrega Adriana Alonso Novais Luiz Gustavo de Almeida Chuffa Debora Aparecida Pires de Campos Zuccari Immunohistochemical Analysis of Inter-Alpha-Trypsin Inhibitor Heavy Chain 2 and Enolase 1 in Canine Mammary Tumors: Associations with Tumor Aggressiveness and Prognostic Significance Veterinary Sciences bitches mammary tumors extracellular vesicles (EVs) biomarkers prognosis |
| title | Immunohistochemical Analysis of Inter-Alpha-Trypsin Inhibitor Heavy Chain 2 and Enolase 1 in Canine Mammary Tumors: Associations with Tumor Aggressiveness and Prognostic Significance |
| title_full | Immunohistochemical Analysis of Inter-Alpha-Trypsin Inhibitor Heavy Chain 2 and Enolase 1 in Canine Mammary Tumors: Associations with Tumor Aggressiveness and Prognostic Significance |
| title_fullStr | Immunohistochemical Analysis of Inter-Alpha-Trypsin Inhibitor Heavy Chain 2 and Enolase 1 in Canine Mammary Tumors: Associations with Tumor Aggressiveness and Prognostic Significance |
| title_full_unstemmed | Immunohistochemical Analysis of Inter-Alpha-Trypsin Inhibitor Heavy Chain 2 and Enolase 1 in Canine Mammary Tumors: Associations with Tumor Aggressiveness and Prognostic Significance |
| title_short | Immunohistochemical Analysis of Inter-Alpha-Trypsin Inhibitor Heavy Chain 2 and Enolase 1 in Canine Mammary Tumors: Associations with Tumor Aggressiveness and Prognostic Significance |
| title_sort | immunohistochemical analysis of inter alpha trypsin inhibitor heavy chain 2 and enolase 1 in canine mammary tumors associations with tumor aggressiveness and prognostic significance |
| topic | bitches mammary tumors extracellular vesicles (EVs) biomarkers prognosis |
| url | https://www.mdpi.com/2306-7381/12/2/110 |
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