Molecular and Cellular Biochemistry

, Volume 440, Issue 1–2, pp 23–31 | Cite as

The overexpression of Hsp90B1 is associated with tumorigenesis of canine mammary glands

  • B. V. Sunil Kumar
  • Rabia Bhardwaj
  • Kanika Mahajan
  • Neeraj Kashyap
  • Ashwani Kumar
  • Ramneek Verma


Heat shock proteins (Hsp) are molecular chaperones that are responsible for protein folding and maintenance of cellular homeostasis. Hsp90, an important member of HSP family, has an important role in breast cancer. Glucose-regulated protein 94 (Grp94) is the endoplasmic reticulum paralog of Hsp90 encoded by Hsp90B1 gene. To test if this protein is overexpressed in dogs with mammary tumor, we estimated and compared its serum levels in healthy dogs and that of dogs with mammary tumors. Hsp90B1 mRNA expression was measured in tumorous and healthy mammary tissues (from age- and breed-matched dogs) by real-time PCR. The gene was found to be overexpressed in mammary tumors (3.586 ± 0.067 times). Further, it was heterologously expressed in a prokaryotic system as 90 kDa protein. A recombinant Grp94-based sandwich ELISA was developed to quantify serum Grp94 in dogs with mammary tumors. Based on receiver-operating characteristics’ analysis, the assay was found to be 90.62% sensitive and 93.75% specific for a cutoff value of 0.35 with respect to histopathological staining in diagnosing the disease. The t test showed that serum Grp94 levels were significantly elevated (92.97 ± 3.62 ng/ml) in dogs with mammary tumors compared with healthy controls (10.30 ± 0.79 ng/ml) (p < 0.0001). These findings suggest that Grp94 might act as a potential biomarker for prognosis of canine mammary tumors and monitoring its therapy.


Hsp90B1 Grp94 Endoplasmic reticulum Expression Canine mammary tumor Prognosis Sandwich ELISA 



The authors are thankful to the Department of Biotechnology, India, for providing the funds to carry out the research work (BT/PR6434/GBD/27/414/2012).

Compliance with ethical standards

Conflict of interest

None of the authors of this paper has a financial or personal relationship with other people or organizations that could inappropriately influence or bias the content of the paper.


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Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  • B. V. Sunil Kumar
    • 1
  • Rabia Bhardwaj
    • 1
  • Kanika Mahajan
    • 1
  • Neeraj Kashyap
    • 2
  • Ashwani Kumar
    • 3
  • Ramneek Verma
    • 1
  1. 1.School of Animal BiotechnologyGuru Angad Dev Veterinary and Animal Sciences UniversityLudhianaIndia
  2. 2.Department of Animal Genetics and Breeding, College of Veterinary SciencesGuru Angad Dev Veterinary and Animal Sciences UniversityLudhianaIndia
  3. 3.Department of Veterinary Surgery and Radiology, College of Veterinary SciencesGuru Angad Dev Veterinary and Animal Sciences UniversityLudhianaIndia

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