Veterinary Research Communications

, Volume 43, Issue 4, pp 231–238 | Cite as

Expression of blood hepatocyte-derived microRNA-122 in canine multicentric lymphoma with hepatic involvement

  • Eman S. RamadanEmail author
  • Adel Abdelbaset Kubesy
  • Taher Ahmed Baraka
  • Faisal Abdelsamad Torad
  • Shaymaa Ismaiel Salem
  • Noha Yousef Salem
Original Article


This study was performed to evaluate the hepatocyte-derived microRNA (miR)-122 as novel diagnostic biomarker in canine lymphoma. Fifteen dogs were enrolled in this study. Dogs presented at Small Animal Teaching Hospital, Faculty of Veterinary Medicine, Cairo University. Dogs were divided into 8 clinically healthy dogs act as control and 7 clinically ill dogs. All dogs were subjected to clinical, ultrasonographic, hemato-biochemical and ultrasound-guided fine-needle biopsy for cytological and histopathological investigations. On the basis of these results, 7 dogs were found to be suffering from multicentric lymphoma involving liver. Serum hepatocyte-derived miRA-122 was determined by real-time quantitative polymerase chain reaction in all dogs. Multicentric lymphoma involving liver manifested by inappetance for several days, depression and peripheral lymphadenopathy. Hematological examination showed significant lymphocytosis. Serum biochemical analysis revealed significant increase in ALT, AST, ALP compared to control dogs. Ultrasonography revealed hypoechoic lymphoid aggregation at area of “porta hepatis” and circumscribed hypoechoic nodule interrupt liver parenchyma. Cytology revealed infiltration of liver tissue by lymphoblast cells and histopathology revealed diffuse infiltration of hepatic sinusoids and portal area by uniform population of small lymphocytes. Serum miRNA-122 analysis showed a significant increase represented as 9.00 fold in canine multicentric lymphoma involving liver. Serum hepatocyte-derived miRNA-122 is of diagnostic value, non invasive, stable and easily measurable blood biomarker for the detection of hepatocellular injury in dogs with multicentric lymphoma involving liver.


Canine Multicentric lymphoma Hepatocyte derived miRNA-122 Cytology Histopathology Ultrasonography 


Funding information

The authors received no financial support for the research, authorship, and/or publication of this article.

Compliance with ethical standards

Conflict of interest

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.


  1. Albonico F, Mortarino M, Avallone G, Gioia G, Comazzi S, Roccabianca P (2013) The expression ratio of miR-17-5p and miR-155 correlates with grading in canine splenic lymphoma. Vet Immunol Immunopathol 155:117–123CrossRefGoogle Scholar
  2. Avery AC, Avery PR (2007) Determining the significance of persistent lymphocytosis. Vet Clin Small Anim 37:267–282CrossRefGoogle Scholar
  3. Baek SJ, McEntee MF, Legendre AM (2009) Review paper: Cancer chemopreventive compounds and canine cancer. Vet Pathol 46:576–588CrossRefGoogle Scholar
  4. Bienzle D, Vernau W (2011) The diagnostic assessment of canine lymphoma: implications for treatment. Clin Lab Med 31:21–39CrossRefGoogle Scholar
  5. Cermelli S, Ruggieri A, Marrero JA, Ioannou GN, Beretta L (2011) Circulating microRNAs in patients with chronic hepatitis C and non-alcoholic fatty liver disease. PLoS One 6(8):923–937CrossRefGoogle Scholar
  6. Cullen JM (2012) Histopathology: Normal liver histology: in Textbbok of canine and feline gastroenterology-SaundersGoogle Scholar
  7. Dirksen K, Verzijl T, van den Ingh TS, Vernooij JC, van der Laan LJ, Burgener IA, Spee B, Fieten H (2016a) Hepatocyte-derived microRNAs as sensitive serum biomarkers of hepatocellular injury in Labrador retrievers. Vet J 211:75–81CrossRefGoogle Scholar
  8. Dirksen K, Verzijl T, Grinwis GC, Favier RP, Penning LC, Burgener IA, van der Laan LJ, Fieten H, Spee B (2016b) Use of serum MicroRNAs as biomarker for hepatobiliary diseases in dogs. J Vet Intern Med 30(6):1816–1823CrossRefGoogle Scholar
  9. Dobson JM, Samuel S, Milstein H, Rogers K, Wood JL (2002) Canine neoplasia in the UK: estimates of incidence rates from a population of insured dogs. J Small Anim Pract 43:240–246CrossRefGoogle Scholar
  10. Dorn CR, Taylor DO, Hibbard HH (1967) Epizootiologic characteristics of canine and feline leukemia and lymphoma. Am J Vet Res 28:993–1001PubMedGoogle Scholar
  11. Edwards DS, Henley WE, Harding EF, Dobson JM, Wood JL (2003) Breed incidence of lymphoma in a UK population of insured dogs. Vet Comp Oncol 1(4):200–206. CrossRefPubMedGoogle Scholar
  12. French TW, Stokol T, Meyer D(2007) Liver. In: Cowell L, Tyler RD, Meinkoth JH, DeNicola DB (eds) diagnostic cytology and hematology of the dog and cat., 3rd ed.Google Scholar
  13. Fulkerson CM (2016) Multicentric Lymphoma in the Dog: An Overview of Diagnostics, Treatment and Prognosis. Available at
  14. Graham MJ, Bell AR, Crewe HK, Moorcraft CL, Walker L, Whittaker EF, Lennard MS (2003) mRNA and protein expression of dog liver cytochromes P450 in relation to the metabolism of human CYP2C substrates. Xenobiotica 33:225–237CrossRefGoogle Scholar
  15. Grow WR (2015) Micro-RNA Profiling for Diagnosis and Prognosis in Canine Multicentric Lymphoma. PhD thesis. The University of Guelph, Guelph, Ontario, CanadaGoogle Scholar
  16. Hume KR, Sylvester SR, Borlle L, Balkman CE, McCleary-Wheeler AL, Pulvino M, Casulo C, Zhao J (2018) Metabolic abnormalities detected in phase II evaluation of doxycycline in dogs with multicentric B-cell lymphoma. Front vet Sci 26:5–25. CrossRefGoogle Scholar
  17. Kerwin SC (1995) Hepatic aspiration and biopsy techniques. Vet Clin North Am Small AnimPract 25:275–291CrossRefGoogle Scholar
  18. Krol J, Loedige I, Filipowicz W (2010) The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet 11:597–610CrossRefGoogle Scholar
  19. Laterza OF, Lim L, Garrett-Engele PW, Vlasakova K, Muniappa N, Tanaka WK, Johnson JM, Sina JF, Fare TL, Sistare FD, Glaab WE (2009) Plasma microRNAs as sensitive and specific biomarkers of tissue injury. Clin Chem 55:1977–1983CrossRefGoogle Scholar
  20. Li S, Zhu J, Fu H, Wan J, Hu Z, Liu S, Li J, Tie Y, Xing R, Zhu J, Sun Z, Zheng X (2012) Hepato-specific microRNA-122 facilitates accumulation of newly synthesized miRNA through regulating PRKRA. Nucleic Acids Res 40:884–891CrossRefGoogle Scholar
  21. Marconato L (2011) The staging and treatment of multicentric high-grade lymphoma in dogs: a review of recent developments and future prospects. The Vet J 188:34–38CrossRefGoogle Scholar
  22. Mattoon J, Nyland T (2015) Small animal diagnostic Ultrasound.3rd ed. Saunders. ElsevierGoogle Scholar
  23. Modiano JF, Breen M, Burnett RC, Parker HG, Inusah S, Thomas R, Avery PR, Lindblad-Toh K, Ostrander EA, Cutter GC, Avery AC (2005) Distinct B-cell and T-cell lymphoproliferative disease prevalence among dog breeds indicates heritable risk. Cancer Res 65:5654–5661CrossRefGoogle Scholar
  24. Mortarino M, Gioia G, Gelain ME, Albonico F, Roccabianca P, Ferri E, Comazzi S (2010) Identification of suitable endogenous controls and differentially expressed microRNAs in canine fresh-frozen and FFPE lymphoma samples. Leuk Res 34:1070–1077CrossRefGoogle Scholar
  25. Mortier F, Daminet S, Vandenabeele S, Van de Maele I (2012) Canine lymphoma: a retrospective study (2009-2010). VLAAMS DIERGENEESKUNDIG TIJDSCHRIFT 81(6):341–351Google Scholar
  26. Nyland TG, Larson MM, Mattoon JS (2015) Liver. In: Mattoon JS and Nyland TG, ed. small animal diagnostic ultrasound. 3rd ed. St. Louis, MO: Elsevier 2015:332–399Google Scholar
  27. Oosthuyzen W, Ten Berg PWL, Francis B, Campbell S, Macklin V, Milne E, Gow AG, Fisher C, Mellanby RJ, Dear JW (2018) Sensitivity and specificity of microRNA-122 for liver disease in dogs. J vet intern Med 32(5):1637–1644. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Owen LN (1980) TNM classification of tumours in domestic animals. 1st ed. Geneva: World Health Organization. Available at
  29. Pastor M, Chalvet-Monfray K, Marchal T, Keck G, Magnol JP (2009) Genetic and environmental risk indicators in canine non-Hodgkin’s lymphomas: breed associations and geographic distribution of 608 cases diagnosed throughout France over 1 year. J Vet Intern Med 23:301–310CrossRefGoogle Scholar
  30. Ponce F, Marchal T, Magnol JP, Turinelli V, Ledieu D, Bonnefont C, Pastor M, Delignette ML, Fournel-Fleury C (2010) A morphological study of 608 cases of canine malignant lymphoma in France with a focus on comparative similarities between canine and human lymphoma morphology. Vet Pathol 47:414–433CrossRefGoogle Scholar
  31. Rothuizen J, Desmet VJ, van der Ingh TSGAM, Twedt DC, Bunch SE, Washabau RJ (2006) Sampling and handling of liver tissue. In: WASVA Liver Standardization Group (ed) WSAVA standards for clinical and histological diagnosis of canine and feline liver disease. Elsevier, Philadelphia, pp 5–14CrossRefGoogle Scholar
  32. Starkey Lewis PJ, Dear J, Platt V, Simpson KJ, Craig DG, Antoine DJ, French NS, Dhaun N, Webb DJ, Costello EM, Neoptolemos JP, Moggs J, Goldring CE, Park BK (2011) Circulating microRNAs as potential markers of human drug-induced liver injury. Hepatology 54:1767–1776CrossRefGoogle Scholar
  33. Stockhaus C, Van Den Ingh T, Rothuizen J, Teske E (2004) A multistep approach in the Cytologic evaluation of liver biopsy samples of dogs with hepatic diseases. Vet Pathol 41:461–470CrossRefGoogle Scholar
  34. Teske E (1994) Canine malignant lymphoma: a review and comparison with human non-hodgkin’s lymphoma. Vet Q 16:209–219CrossRefGoogle Scholar
  35. Teske E, de Vos JP, Egberink HF, Vos JH (1994) Clustering in canine malignant lymphoma. Vet Q 16:134–136CrossRefGoogle Scholar
  36. Thangapandiyan M, Balachandran C, MuraliManohar B (2013) Incidence and haemato-biochemical changes in canine lymphoma. Tamilnadu JVet Anim Sci 9(1):29–31Google Scholar
  37. Uhl E, Krimer P, Schliekelman P, Tompkins SM, Suter S (2011) Identification of altered MicroRNA expression in canine lymphoid cell lines and cases of B-and T-cell lymphomas. Genes Chromosom Cancer 50:950–967CrossRefGoogle Scholar
  38. Vail DM, 2017. Hematopoietic tumors. Textbook of veterinary internal medicine: diseases of the dog and cat. Ettinger SJ, Feldman EC, Côté E. 8th edn, ElsevierGoogle Scholar
  39. Vail DM, Young KM (2007) Hematopoietic tumors. In Withrow SJ , Vail DM (eds) Textbook of Withrow & MacEwen’s Small Animal Clinical Oncology.4th ed. Saunders.St. Louis , p: 699–733Google Scholar
  40. Vail DM, Pinkerton ME, Young KM (2013) Hematopoietic tumors: canine lymphoma and lymphoid leukemias. 5th ed In: Withrow SJ, Vail DM, Page RL, editors. , editors. Withrow & MacEwen’s Small Animal Clinical Oncology. St. Louis, MO: Elsevier Saunders, 612 pGoogle Scholar
  41. Van der Meer AJ, Farid WR, Sonneveld MJ, de Ruiter PE, Boonstra A, van Vuuren AJ, Verheij J, Hansen BE, de Knegt RJ, van der Laan LJ, Janssen HL (2013) Sensitive detection of hepatocellular injury in chronic hepatitis C patients with circulating hepatocyte-derived microRNA-122. J Viral Hepat 20:158–166CrossRefGoogle Scholar
  42. vanSteenbeek FG, Van den Bossche L, Grinwis GC, Kummeling A, van Gils IH, Koerkamp MJ, van Leenen D, Holstege FC, Penning LC, Rothuizen J, Leegwater PA, Spee B (2013) Aberrant gene expression in dogs with portosystemic shunts. PLoS One 8:657–662Google Scholar
  43. Vezzali E, Parodi AL, Marcato PS, Bettini G (2010) Histopathologic classification of 171 cases of canine and feline nonhodgkin lymphoma according to the WHO. Vet Comp Oncol 8:38–49CrossRefGoogle Scholar
  44. Villamil JA, Henry CJ, Hahn AW, Bryan JN, Tyler JW, Caldwell CW (2009) Hormonal and sex impact on the epidemiology of canine lymphoma. J Cancer Epidemiol 2009:1–7. CrossRefGoogle Scholar
  45. Wang K, Zhang S, Marzolf B, Troisch P, Brightman A, Hu Z, Hood LE, Galas DJ (2009) Circulating microRNAs, potential biomarkers for drug-induced liver injury. Proc Natl Acad Sci U S A 106(11):4402–4407. CrossRefPubMedPubMedCentralGoogle Scholar
  46. Weiss DJ, Moritz A (2002) Liver cytology. Vet Clin Small Anim 32:1267–1291CrossRefGoogle Scholar
  47. Williams MJ, Avery AC, Lana SE, Hillers KR, Bachand AM, Avery PR (2008) Canine lymphoproliferative disease characterized by lymphocytosis: Immunophenotypic markers of prognosis. J Vet Intern Med 22:596–601CrossRefGoogle Scholar
  48. Yuan JS, Reed A, Chen F, Stewart CN Jr (2006) Statistical analysis of real-time PCR data. BMC Bioinf 7:85CrossRefGoogle Scholar
  49. Zandvliet M (2016) Canine lymphoma: a review. Vet Q 36(2):76–104CrossRefGoogle Scholar
  50. Zhang Y, Jia Y, Zheng R, Guo Y, Wang Y, Guo H, Fei M, Sun S (2010) Plasma microRNA-122 as a biomarker for viral-, alcohol-,and chemical-related hepatic diseases. ClinChem 56:1830–1838Google Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Internal Medicine and Infectious Diseases, Faculty of Veterinary MedicineCairo UniversityGizaEgypt
  2. 2.Department of Surgery and Anesthesia, Faculty of Veterinary MedicineCairo UniversityGizaEgypt
  3. 3.Department of Clinical Pathology, Faculty of Veterinary MedicineCairo UniversityGizaEgypt

Personalised recommendations