Clinical relevance of neutral endopeptidase (NEP/CD10) in melanoma.
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.


CONCLUSIONS
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.
Velazquez
Elsa F
EF
Ronald O. Perelman Department of Dermatology, New York University School of Medicine, New York, NY, USA. efvelazquez@partners.org
Yancovitz
Molly
M
Pavlick
Anna
A
Berman
Russell
R
Shapiro
Richard
R
Bogunovic
Dusan
D
O'Neill
David
D
Yu
Yi-Lo
YL
Spira
Joanna
J
Christos
Paul J
PJ
Zhou
Xi Kathy
XK
Mazumdar
Madhu
M
Nanus
David M
DM
Liebes
Leonard
L
Bhardwaj
Nina
N
Polsky
David
D
Osman
Iman
I
D045744:Cell Line, Tumor; D015536:Down-Regulation / Q000235:genetics; D023381:Endpoint Determination; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D052138:Genes, Neoplasm; D006801:Humans; D007150:Immunohistochemistry; D008545:Melanoma / Q000201:enzymology* / Q000235:genetics; D009362:Neoplasm Metastasis; D015260:Neprilysin / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016019:Survival Analysis
D012333:RNA, Messenger; D015260:Neprilysin