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Research Article

volume 39 issue 4 (december 2019) : 335-340,   Doi: 10.18805/ag.D-4968

A Study on Immunohistochemical Expression of Progesterone Receptor and Cell Proliferation by AgNOR in Canine Mammary Tumors (CMT)

P
P.L. Leena Rajathy
K
K.C. Varshney
M
M.G. Nair
A
A.W. Lakkawar
B
B. Ramesh Kumar
1Department of Veterinary Surgery and Radiology, Rajiv Gandhi Institute of Veterinary Education and Research (RIVER), Kurumbapet, Pondicherry-605 009, India.
Cite article:- Rajathy Leena P.L., Varshney K.C., Nair M.G., Lakkawar A.W., Kumar Ramesh B. (2019). A Study on Immunohistochemical Expression of Progesterone Receptor and Cell Proliferation by AgNOR in Canine Mammary Tumors (CMT). Agricultural Science Digest. 39(4): 335-340. doi: 10.18805/ag.D-4968.

ABSTRACT

Mammary gland tumors are the most commonly diagnosed tumors in domestic dogs. Although immunohistochemical methods provide valuable information such as the location and semi-quantitative data of the interested antigens in particular tumors, conventional methods like histopathological diagnosis remain useful and necessary for identification and classification of tumors. In the present study, we combined histopathology with immunohistochemical staining for progesterone receptor (PR) and special staining argyrophilic nucleolar organizer regions (AgNOR) for cell proliferation in canine mammary gland tumors. Twenty-nine dogs with primary mammary gland neoplasms (11 benign and 18 malignant) were included in this study. All 29 tumors were stained with AgNOR but only 15 cases expressed positive PR signals in the nuclei of neoplastic alveolar and ductal epithelial cells. A weak positive correlation (r = 0.2) was observed between PR and AgNOR index of benign tumors and a very weak negative correlation (r = -0.1833) between PR and AgNOR index of malignant tumors suggesting that higher cellular proliferation is associated with low expression of progesterone receptors. 

REFERENCES

  1. Anil, A. (2007). Identification of stem cell lineage cd44+/cd24- and apoptotic marker in mammary tumours of dogs. M.V.Sc. Thesis, TANUVAS, Chennai, India.
  2. Bostock, D.E. (1986). Canine and feline mammary neoplasms. The British Veterinary Journal. 112: 506-15.
  3. Bronden, L.B., Nielsen, S.S., Toft, N., Kristensen, A.T. (2010). Data from the Danish veterinary cancer registry on the occurrence and distribution of neoplasms in dogs in Denmark. The Veterinary record. 166(19):586–90.
  4. Chang, C.C., Tsai, M.H., Liao, J.W. (2009). Evaluation of hormone receptor expression for use in predicting survival of female dogs with malignant mammary gland tumors. Journal of the American Veterinary Medical Association. 235:391–396.
  5. Dorn, C.R., Taylor, D.O.N., Schneider, R., Hibbard, H.H. and Klauber, M.R. (1968). Survey of animal neoplasms in Alameda and Contra Costa Countries, Californis. II. Cancer morbidity in dogs and cats from Alameda Country. Journal of the National Cancer Institute. 40: 307-318. 
  6. Fowler, J. and Cohen, L. (1990). Practical Statistics for Field Biology. John Wiley & Sons, England.
  7. Geraldes, M., Gartner, F. and Schmitt, F. (2000).Immunohistochemical study of hormonal receptors and cell proliferation in normal canine mammary glands and spontaneous mammary tumours. Veterinary Record. 146: 403-406. 
  8. Graham, J.C., O’Keefe, D.A. and Gelberg, H.B. (1999). Immuno    histochemical assay for detecting estrogen receptors in canine mammary tumors. American journal of veterinary research. 60:627–630.
  9. Hampe, I.F. and Misdorp, W. (1974). Tumors and dysplasias of the mammary gland. Bulletin of the WHO. 50: 111-133.
  10. Jain, V. and Ragunath, M. (2007). Incidence of canine mammary neoplasms in and around Ludhiana. Punjab Veterinary Journal. 5: 42-43.
  11. Jelesijevic, T., Jovanovic, M., Knezevic Milijana and Aleksic-    Kovacevic sanja (2003). Quantitative and qualitative Analysis of AgNOR in Benign and Malignant canine Mammary Gland Tumors. Acta Veterinaria. 53: 353-360.
  12. Juntes, P. and Pogacnik, M. (2000). Morphometric analysis of AgNORs in tubular and papillary parts of canine mammary tumors. Veterinary Pathology. 22:185–192. 
  13. Kaszak, I., Ruszczak, A., Kanafa, S., Kacprzak, K., Król, M. and Jurka, P. (2018). Current biomarkers of canine mammary tumors. Acta Veterinaria Scandinavica. 60:66.
  14. Kumar, P., Kumar, R., Pawaiya, R.V.S., Madhu, B.P. (2010).Diagnostic significance of mitotic index and AgNOR count in canine mammary tumours. Brazilian Journal of Veterinary Pathology. 3: 41–45.
  15. Lohr, C.V., Teifke, J.P., Failing, K. and Weiss (1997). Characterization of the proliferation state in canine mammary tumors by the standardized AgNOR method with postfixation and immunohistologic detection of Ki-67 and PCNA. Veterinary Pathology. 34: 212-221.
  16. Luna, L.G. (1968). Manual of histologic staining methods of the Armed Forces Institute of Pathology. Mc Graw Hill, New York, pp 32-46.
  17. Madewell, B.R. (2001). Cellular proliferation in tumors: A review of methods, interpretation, and clinical applications. Journal of Veterinary Internal Medicine. 15: 334-340.
  18. Martin de las Mulas, J., Millan, M.Y. and Dios, R. (2005). A prospective analysis of immunohistochemically determined estrogen receptor á and progesterone receptor expression and host and tumor factors as predictors of disease-free period in mammary tumors of the dog. Veterinary Pathology. 42: 200-212.
  19. Millanta, F., Calandrella, M., Bari, G., Niccolini, M., Vannozzi, I. and Poli, A. (2005). Comparison of steroid receptor expression in normal, dysplastic, and neoplastic canine and feline mammary tissues. Research in Veterinary Science. 79: 225-232.
  20. Mitchell, L., de la Iglesia, F.A., Wenkoff, M.S., van Dreumel, A.A. and Lumb, G. (1974). Mammary tumors in dogs: survey of clinical and pathological characteristics. Canadian Veterinary Journal. 15: 131-138.
  21. Moulton, J.E. (1990).Tumors of the mammary gland. In: Tumors in Domestic Animals. 3rd edn. (Ed.) J E. Moulton. Berkely, University of California Press. 
  22. Mohr, A., Lüder Ripoli, F., Hammer, S.C., et al. (2016). Hormone Receptor Expression Analyses in Neoplastic and Non-Neoplastic Canine Mammary Tissue by a Bead Based Multiplex Branched DNA Assay: A Gene Expression Study in Fresh Frozen and Formalin-Fixed, Paraffin-Embedded Samples.PLOS One. 11(9):e0163311. 
  23. Munakata, S. and Hendricks, J.B. (1994). A multilabeling technique for simultaneous demonstration and quantitation of Ki-    67 and nucleolar organizer regions (AgNORs) in paraffin-    embedded tissue. Journal of Histochemistry and Cytochemistry. 42: 789-793.
  24. Nieto, A., Pena, L. and Perez-Alenza, M.D. (2000).Immunohistologic detection of estrogen receptor alpha in canine mammary tumors: clinical and pathologic associations and prognostic significance. Veterinary Pathology. 37:239–247.
  25. Priya, S. (2005). Comparison of nipple aspirate fluid cytology with other routine tests in the diagnosis of canine mammary tumours. M.V.Sc thesis. TANUVAS, Chennai, India.
  26. Reddy, G.B.M., Kumar, R., Sharma, A.K. and Ravindran, R. (2007). Canine mammary tumors: study on cell proliferative markers. Proceedings of XXIV Annual Conference of Veterinary pathologists. Oct. 1-3, Tirupati, India.
  27. Toniti, W., Buranasinsup, S., Kongcharoen, A., Charoonrut, P., Puchadapirom, P., and Kasorndorkbua, C. (2009). Immunohistochemical Determination of Estrogen and Progesterone Receptors in Canine Mammary Tumors. Asian Pacific Journal of Cancer Prevention. 10: 907-911.
  28. Thuroczy, J., Reisvaag, G.J, K., Perge, E., Tibold, A., Szilagyi, J. and Balogh, L. (2007). Immunohistochemical detection of Progesterone and Cellular Proliferation in Canine Mammary Tumors. Journal of Comparative Pathology. 137: 122-129. 
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    Research Article

    volume 39 issue 4 (december 2019) : 335-340,   Doi: 10.18805/ag.D-4968

    A Study on Immunohistochemical Expression of Progesterone Receptor and Cell Proliferation by AgNOR in Canine Mammary Tumors (CMT)

    P
    P.L. Leena Rajathy
    K
    K.C. Varshney
    M
    M.G. Nair
    A
    A.W. Lakkawar
    B
    B. Ramesh Kumar
    1Department of Veterinary Surgery and Radiology, Rajiv Gandhi Institute of Veterinary Education and Research (RIVER), Kurumbapet, Pondicherry-605 009, India.
    Cite article:- Rajathy Leena P.L., Varshney K.C., Nair M.G., Lakkawar A.W., Kumar Ramesh B. (2019). A Study on Immunohistochemical Expression of Progesterone Receptor and Cell Proliferation by AgNOR in Canine Mammary Tumors (CMT). Agricultural Science Digest. 39(4): 335-340. doi: 10.18805/ag.D-4968.

    ABSTRACT

    Mammary gland tumors are the most commonly diagnosed tumors in domestic dogs. Although immunohistochemical methods provide valuable information such as the location and semi-quantitative data of the interested antigens in particular tumors, conventional methods like histopathological diagnosis remain useful and necessary for identification and classification of tumors. In the present study, we combined histopathology with immunohistochemical staining for progesterone receptor (PR) and special staining argyrophilic nucleolar organizer regions (AgNOR) for cell proliferation in canine mammary gland tumors. Twenty-nine dogs with primary mammary gland neoplasms (11 benign and 18 malignant) were included in this study. All 29 tumors were stained with AgNOR but only 15 cases expressed positive PR signals in the nuclei of neoplastic alveolar and ductal epithelial cells. A weak positive correlation (r = 0.2) was observed between PR and AgNOR index of benign tumors and a very weak negative correlation (r = -0.1833) between PR and AgNOR index of malignant tumors suggesting that higher cellular proliferation is associated with low expression of progesterone receptors. 

    REFERENCES

    1. Anil, A. (2007). Identification of stem cell lineage cd44+/cd24- and apoptotic marker in mammary tumours of dogs. M.V.Sc. Thesis, TANUVAS, Chennai, India.
    2. Bostock, D.E. (1986). Canine and feline mammary neoplasms. The British Veterinary Journal. 112: 506-15.
    3. Bronden, L.B., Nielsen, S.S., Toft, N., Kristensen, A.T. (2010). Data from the Danish veterinary cancer registry on the occurrence and distribution of neoplasms in dogs in Denmark. The Veterinary record. 166(19):586–90.
    4. Chang, C.C., Tsai, M.H., Liao, J.W. (2009). Evaluation of hormone receptor expression for use in predicting survival of female dogs with malignant mammary gland tumors. Journal of the American Veterinary Medical Association. 235:391–396.
    5. Dorn, C.R., Taylor, D.O.N., Schneider, R., Hibbard, H.H. and Klauber, M.R. (1968). Survey of animal neoplasms in Alameda and Contra Costa Countries, Californis. II. Cancer morbidity in dogs and cats from Alameda Country. Journal of the National Cancer Institute. 40: 307-318. 
    6. Fowler, J. and Cohen, L. (1990). Practical Statistics for Field Biology. John Wiley & Sons, England.
    7. Geraldes, M., Gartner, F. and Schmitt, F. (2000).Immunohistochemical study of hormonal receptors and cell proliferation in normal canine mammary glands and spontaneous mammary tumours. Veterinary Record. 146: 403-406. 
    8. Graham, J.C., O’Keefe, D.A. and Gelberg, H.B. (1999). Immuno    histochemical assay for detecting estrogen receptors in canine mammary tumors. American journal of veterinary research. 60:627–630.
    9. Hampe, I.F. and Misdorp, W. (1974). Tumors and dysplasias of the mammary gland. Bulletin of the WHO. 50: 111-133.
    10. Jain, V. and Ragunath, M. (2007). Incidence of canine mammary neoplasms in and around Ludhiana. Punjab Veterinary Journal. 5: 42-43.
    11. Jelesijevic, T., Jovanovic, M., Knezevic Milijana and Aleksic-    Kovacevic sanja (2003). Quantitative and qualitative Analysis of AgNOR in Benign and Malignant canine Mammary Gland Tumors. Acta Veterinaria. 53: 353-360.
    12. Juntes, P. and Pogacnik, M. (2000). Morphometric analysis of AgNORs in tubular and papillary parts of canine mammary tumors. Veterinary Pathology. 22:185–192. 
    13. Kaszak, I., Ruszczak, A., Kanafa, S., Kacprzak, K., Król, M. and Jurka, P. (2018). Current biomarkers of canine mammary tumors. Acta Veterinaria Scandinavica. 60:66.
    14. Kumar, P., Kumar, R., Pawaiya, R.V.S., Madhu, B.P. (2010).Diagnostic significance of mitotic index and AgNOR count in canine mammary tumours. Brazilian Journal of Veterinary Pathology. 3: 41–45.
    15. Lohr, C.V., Teifke, J.P., Failing, K. and Weiss (1997). Characterization of the proliferation state in canine mammary tumors by the standardized AgNOR method with postfixation and immunohistologic detection of Ki-67 and PCNA. Veterinary Pathology. 34: 212-221.
    16. Luna, L.G. (1968). Manual of histologic staining methods of the Armed Forces Institute of Pathology. Mc Graw Hill, New York, pp 32-46.
    17. Madewell, B.R. (2001). Cellular proliferation in tumors: A review of methods, interpretation, and clinical applications. Journal of Veterinary Internal Medicine. 15: 334-340.
    18. Martin de las Mulas, J., Millan, M.Y. and Dios, R. (2005). A prospective analysis of immunohistochemically determined estrogen receptor á and progesterone receptor expression and host and tumor factors as predictors of disease-free period in mammary tumors of the dog. Veterinary Pathology. 42: 200-212.
    19. Millanta, F., Calandrella, M., Bari, G., Niccolini, M., Vannozzi, I. and Poli, A. (2005). Comparison of steroid receptor expression in normal, dysplastic, and neoplastic canine and feline mammary tissues. Research in Veterinary Science. 79: 225-232.
    20. Mitchell, L., de la Iglesia, F.A., Wenkoff, M.S., van Dreumel, A.A. and Lumb, G. (1974). Mammary tumors in dogs: survey of clinical and pathological characteristics. Canadian Veterinary Journal. 15: 131-138.
    21. Moulton, J.E. (1990).Tumors of the mammary gland. In: Tumors in Domestic Animals. 3rd edn. (Ed.) J E. Moulton. Berkely, University of California Press. 
    22. Mohr, A., Lüder Ripoli, F., Hammer, S.C., et al. (2016). Hormone Receptor Expression Analyses in Neoplastic and Non-Neoplastic Canine Mammary Tissue by a Bead Based Multiplex Branched DNA Assay: A Gene Expression Study in Fresh Frozen and Formalin-Fixed, Paraffin-Embedded Samples.PLOS One. 11(9):e0163311. 
    23. Munakata, S. and Hendricks, J.B. (1994). A multilabeling technique for simultaneous demonstration and quantitation of Ki-    67 and nucleolar organizer regions (AgNORs) in paraffin-    embedded tissue. Journal of Histochemistry and Cytochemistry. 42: 789-793.
    24. Nieto, A., Pena, L. and Perez-Alenza, M.D. (2000).Immunohistologic detection of estrogen receptor alpha in canine mammary tumors: clinical and pathologic associations and prognostic significance. Veterinary Pathology. 37:239–247.
    25. Priya, S. (2005). Comparison of nipple aspirate fluid cytology with other routine tests in the diagnosis of canine mammary tumours. M.V.Sc thesis. TANUVAS, Chennai, India.
    26. Reddy, G.B.M., Kumar, R., Sharma, A.K. and Ravindran, R. (2007). Canine mammary tumors: study on cell proliferative markers. Proceedings of XXIV Annual Conference of Veterinary pathologists. Oct. 1-3, Tirupati, India.
    27. Toniti, W., Buranasinsup, S., Kongcharoen, A., Charoonrut, P., Puchadapirom, P., and Kasorndorkbua, C. (2009). Immunohistochemical Determination of Estrogen and Progesterone Receptors in Canine Mammary Tumors. Asian Pacific Journal of Cancer Prevention. 10: 907-911.
    28. Thuroczy, J., Reisvaag, G.J, K., Perge, E., Tibold, A., Szilagyi, J. and Balogh, L. (2007). Immunohistochemical detection of Progesterone and Cellular Proliferation in Canine Mammary Tumors. Journal of Comparative Pathology. 137: 122-129. 
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