Abstract
BACKGROUND: Mammary tumors are the most common tumors in female dogs. An early diagnosis makes the treatment easier. AIMS: The present study aimed to assess heat shock protein D1 (HSPD1) expression in canine mammary tumors. METHODS: Canine mammary tumor (CMT) samples were collected from clinics after surgery. Expression of HSPD1 transcript in CMT and apparently healthy mammary tissues was analyzed by SYBR green based real-time PCR (qRT-PCR). Further, gene encoding the immuno-dominant region of HSPD1 was cloned using the expression vector pPROEX-HTa and expressed in a prokaryotic system and recombinant HSPD1 (rHSPD1) was purified by affinity chromatography. Hyperimmune serum was raised against rHSPD1 in mice, and immunohistochemistry was standardized to assess the expression of this protein in various histotypes of canine mammary tumors. RESULTS: An elevated HSPD1 mRNA expression (5.973 ± 0.862 folds) was observed in canine mammary tumors. Upon purification, a 60 kDa recombinant protein was obtained and confirmed by Western blotting. In 83.3% of healthy mammary tissues, a mild/feeble HSPD1 expression was observed whereas, a strong expression of HSPD1 was seen in 80% CMT samples. CONCLUSION: The findings suggested that HSPD1 could be used as a molecular marker for canine mammary tumors.