Research

Clinical relevance of Neutral Endopeptidase (NEP/CD10) in melanoma

Elsa F Velazquez^{* 1,6} , Molly Yancovitz^{* 1} , Anna Pavlick^1,2 , Russell Berman³ , Richard Shapiro³ , Dusan Bogunovic² , David O'Neill² , Yi-Lo Yu² , Joanna Spira¹ , Paul J Christos⁴ , Xi Kathy Zhou⁴ , Madhu Mazumdar⁴ , David M Nanus⁵ , Leonard Liebes² , Nina Bhardwaj^1,2 , David Polsky¹ and Iman Osman^{* 1,2}

¹Ronald O. Perelman Department of Dermatology, New York University School of Medicine, New York, NY, USA

²Department of Medicine, New York University School of Medicine, New York, NY, USA

³Department of Surgery, New York University School of Medicine, New York, NY, USA

⁴Department of Public Health, Weill Medical College of Cornell University, New York, NY, USA

⁵Department of Medicine, Weill Medical College of Cornell University, New York, NY, USA

⁶Department of Pathology, Brigham and Women's Hospital, Harvard School of Medicine, Boston, MA, USA

author email corresponding author email* Contributed equally

Journal of Translational Medicine 2007, 5:2doi:10.1186/1479-5876-5-2

Published:	5 January 2007

Abstract

Background

Overexpression of Neutral Endopeptidase (NEP) has been reported in metastatic carcinomas, implicating NEP in tumor progression and suggesting a role for NEP inhibitors in its treatment. We investigated the role of NEP expression in the clinical progression of cutaneous melanoma.

Methods

We screened 7 melanoma cell lines for NEP protein expression. NEP-specific siRNA was transfected into the lines to examine the role of gene transcription in NEP expression. Immunohistochemistry was done for 93 specimens and correlated with clinicopathologic parameters. Thirty-seven metastatic melanoma specimens were examined for NEP transcript expression using Affymetrix GeneChips. In a subset of 25 specimens for which both transcript and protein expression was available, expression ratios were used to identify genes that co-express with NEP in GeneChip analysis.

Results

NEP was overexpressed in 4/7 human melanoma cell lines, and siRNA knock-down of NEP transcripts led to downregulation of its protein expression. NEP protein overexpression was significantly more common in metastatic versus primary tumors (P = 0.002). Twelve of 37 (32%) metastatic tumors had increased NEP transcript expression, and an association was observed between NEP transcript upregulation and protein overexpression (P < 0.0001). Thirty-eight genes were found to significantly co-express with NEP (p < 0.005). Thirty-three genes positively correlated with NEP, including genes involved in the MAP kinase pathway, antigen processing and presentation, apoptosis, and WNT signaling pathway, and 5 genes negatively correlated with NEP, including genes of focal adhesion and the notch signaling pathways.

Conclusion

NEP overexpression, which seems to be largely driven by increased transcription, is rare in primary melanoma and occurs late in melanoma progression. Functional studies are needed to better understand the mechanisms of NEP regulation in melanoma.