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Short Communication · Volume 4, Issue 7, Pages 342–345, 4 July 2016 · Open Access

Development of Real-Time PCR Assays for Detecting Matrix Metalloproteinases-2 & 9 Over-expression in Canine Mammary Tumours

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Journal:
Advances in Animal and Veterinary Sciences
Published:
4 July 2016
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From the article

  • Development of Real-Time PCR Assays for Detecting Matrix Metalloproteinases-2 & 9 Over-expression in Canine Mammary Tumours.
  • The over-expression of MMP2 and MMP9 was examined in 12 CMT tissues by real-time PCR analysis.
  • Over-expression of MMP9 was detected in 5 out of 12(50%) tissues of canine mammary tumour.
  • The samples from CMT cases showed 2.23±0.10 to 29.80±3.5 fold higher expression levels of MMP2 as compared to normal healthy mammary gland tissue of dog.

Selected from the article's own text.

Abstract

 

Sonal Saxena1*, Sameer Shrivastava1, Richa Arora1, Shahid Hussain1, Subas Chandra Jena1, Manoj Kumar1, Resma Kochumanayin Vasu1, Saumya Srivastava1, Priyanka Sharma1, Naveen Kumar2, Kuldeep Dhama3

 

1Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, U.P., India; 2Division of Veterinary Surgery, Indian Veterinary Research Institute, Izatnagar, U.P., India; 3Division of Pathology, Indian Veterinary Research Institute, Izatnagar, U.P., India.

 

Abstract | Matrix Metalloproteinases (MMP) 2 & 9 are overexpressed in wide variety of cancers and play important roles in tumour cell proliferation, invasion and metastasis. Tissue and serum levels of MMP2 and MMP9 correlate with disease prognosis. Real-time PCR is emerging as an alternative or supplementary technique to immunohistochemistry (IHC) for sensitive detection of tumour markers in tissues. Therefore, in this study real-time PCR assays were standardized for detecting over-expression of MMP2 and MMP9 in canine mammary tumour (CMT). Primers designed for real-time PCR analysis of MMP2 & MMP9 were specific as indicated by amplification plots and melt-curve analysis of amplicons. These amplicons showed sharply defined melting curves with single peaks at expected melting points. The over-expression of MMP2 and MMP9 was examined in 12 CMT tissues by real-time PCR analysis. Over-expression of MMP2 & MMP9 was observed in 58.3(7/12) and 41.6 % (5/12) cases of CMT respectively. Expression levels of MMP2 were 2.23±0.10 to 29.80±3.5 fold higher and for MMP2, 1.5±0.56 to 22.67±2.35 fold higher in cases of CMT than normal mammary gland biopsy. Approximately 33.3 % (4/12) of CMT tissues showed co-expression of both MMP2 and MMP9. Further studies are required to correlate MMP2 and MMP9 expression levels with CMT prognosis.

 

Keywords | MMP2, MMP9, Matrix-metalloproteinase, Real-time PCR, Canine mammary tumour (CMT)


Acknowledgements

The authors are thankful to Director, IVRI, and Izatnagar for providing necessary infrastructure and facilities to carry out this research work. Authors are also thankful to Department of Biotechnology (DBT), Govt. of India (Project grant No. BT/PR13900/ADV/57/44/2010) and Indian Council of Agricultural Research (ICAR) NAE programme on biosensors for supporting the study by research grants.

Conflict of Interests

Authors of this manuscript declare no conflict of interest.

authors’ contribution

S. Saxena & S. Shrivastava designed the work, conducted real-time PCR experiments and drafted the manuscript. R. Arora, M. Kumar, R.K. Vasu & S. Srivastava isolated RNA from tumour tissues, prepared cDNA and helped in standardization of real time PCR. S. C. Jena, and P. Sharma helped in data analysis. N. Kumar collected tumour tissues from surgically operated CMT cases. K. Dhama helped in data analysis and drafting the manuscript.

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Development of Real-Time PCR Assays for Detecting Matrix Metalloproteinases-2 & 9 Over-expression in Canine Mammary Tumours · ResearchersLinks