Aminopeptidase N (APN; EC 3.4.11.2) is a membrane dimeric metallopeptidase involved in differentiation, development, and proliferative processes of several tissues. Recent studies have demonstrated the increased expression and activity of this enzyme in several cancers. However, there are no available data about the impact of this peptidase in the biological aggressiveness and the survival of colorectal cancer (CRC) patients.
Methods
The activity and mRNA expression of APN in tumor tissue (n = 81) and plasma (n = 40) of patients with CRC of low and high grades and stages were prospectively analyzed by fluorimetric and quantitative reverse transcriptase–polymerase chain reaction methods. Data obtained in adenoma and CRC were compared with those from the surrounding normal mucosa. Classic clinical and pathological parameters were stratified following APN data and analyzed for 5-year survival.
Results
mRNA levels of APN (ANPEP) were lower in colorectal adenomas and adenocarcinomas than in the surrounding uninvolved mucosa (Kruskal-Wallis, P < 0.001). Aminopeptidase N activity in CRC tissue was higher in patients with better overall survival (log-rank P < 0.05, Cox analysis P < 0.05). By contrast, higher plasmatic APN activity correlated with worse overall survival (log-rank P < 0.01, Cox analysis P < 0.05).
Conclusions
Aminopeptidase N activity in tissue and plasma from CRC patients is an independent prognostic factor of 5-year survival. The determination of APN activity levels in the plasma may be a safe, minimally invasive, and inexpensive way to define the aggressiveness of CRC in daily practice.
PetrovicN, SchackeW, ShapiroLH.CD13/aminopeptidase N in tumor growth and angiogenesis. In:HooperNM, LendeckelU, eds.Aminopeptidases in Biology and Disease. New York: Kluwer Academic/Plenum Press; 2004:179–200.
8.
Carl-McGrathS, LendeckelU, EbertM, . Ectopeptidases in tumor biology: a review. Histol Histopathol. 2006; 21: 1339–1353.
9.
CorderoOJ, AyudeD, NogueiraM, . Preoperative serum CD26 levels: diagnostic efficiency and predictive value for colorectal cancer. Br J Cancer. 2000; 80: 1139–1146.
10.
HashidaH, TakabayashiA, KanaiM, . Aminopeptidase N is involved in cell motility and angiogenesis: its clinical significance in human colon cancer. Gastroenterology. 2002; 122: 376–386.
11.
ZhangQ, WangJ, ZhangH, . Expression and clinical significance of aminopeptidase N/CD13 in non-small cell lung cancer. J Cancer Res Ther. 2015; 11: 223–228.
12.
MurakamiH, YokoyamaA, KondoK, . Circulating aminopeptidase N/CD13 is an independent prognostic factor in patients with non-small cell lung cancer. Clin Cancer Res. 2005; 11: 8674–8679.
13.
TokuharaT, HattoriN, IshidaH, . Clinical significance os aminopeptidase N in non–small cell lung cancer. Clin Cancer Res. 2006; 12: 3971–3978.
14.
LarrinagaG, BlancoL, SanzB, . The impact of peptidase activity in clear cell renal cell carcinoma survival. Am J Physiol Renal Physiol. 2012; 303: F1584–F1591.
15.
LarrinagaG, PerezI, BlancoL, . Increased prolyl endopeptidase activity in human neoplasia. Regul Pept. 2010; 163: 102–106.
16.
LarrinagaG, BlancoL, ErrarteP, . Altered peptidase activities in thyroid neoplasia and hyperplasia. Dis Markers. 2013; 35 (6): 825–832.
17.
BlancoL, LarrinagaG, PerezI, . Acid, basic, and neutral peptidases present different profiles in chromophobe renal cell carcinoma and in oncocytoma. Am J Physiol Renal Physiol. 2008; 294: F850–F858.
18.
RealFX, XuM, ViláMR, . Intestinal brush-border–associated enzymes: co-ordinated expression in colorectal cancer. Int J Cancer. 1992; 51 (2): 173–181.
19.
Mina-OsorioP.The moonlighting enzyme CD13: old and new functions to target. Trends Mol Med. 2008; 14 (8): 361–371.
20.
FavaloroEJ, BrowningT, FaceyD.CD13 (GP150; aminopeptidase-N): predominant functional activity in blood is localized to plasma and is not cell-surface associated. Exp Hematol. 1993; 21: 1695–1701.
21.
JungK, PergandeM, WischkeUW.Characterization of particulate and soluble variants of the brush-border enzymes alanine aminopeptidase, alkaline phosphatase and gammaglutamyltransferase in human urine. Biomed Biochim Acta. 1984; 43: 1357–1364.
22.
van HensbergenY, BroxtermanHJ, HanemaaijerR, . Soluble aminopeptidase N/CD13 in malignant and nonmalignant effusions and intratumoral fluid. Clin Cancer Res. 2002; 8 (12): 3747–3754.
23.
WickströmM, LarssonR, NygrenP, . Aminopeptidase N (CD13) as a target for cancer chemotherapy. Cancer Sci. 2011; 102: 501–508.
24.
ZucaliPA, SimonelliM, De VincenzoF, . Phase I and pharmacodynamic study of high-dose NGR-hTNF in patients with refractory solid tumours. Br J Cancer. 2013; 108: 58–63.
25.
IkedaN, NakajimaY, TokuharaT, . Clinical significance of aminopeptidase N/CD13 expression in human pancreatic carcinoma. Clin Cancer Res. 2003; 9 (4): 1503–1508.
26.
KawamuraJ, ShimadaY, KitaichiH, . Clinicopathologic significance of aminopeptidase N/CD13 expression in human gastric carcinoma. Hepatogastroenterology. 2007; 54(73): 36–40.
27.
LarrinagaG, PerezI, ArizU, . Clinical impact of aspartyl aminopeptidase expression and activity in colorectal cancer. Transl Res. 2013; 162: 297–308.
28.
LarrinagaG, PerezI, BlancoL, . Prolyl endopeptidase activity is correlated with colorectal cancer prognosis. Int J Med Sci. 2013; 11: 199–208.
29.
EdgeSB, ByrdDR, ComptonCC, ., eds.AJCC Cancer Staging Manual. New York: Springer; 2010.
30.
WashingtonMK, BerlinJ, BrantonP, . Protocol for the examination of specimens from patients with primary carcinoma of the colon and rectum. Arch Pathol Lab Med. 2009; 133: 1539–1551.
31.
OlivoRA, TeixeiraPT, SilveiraPF, . Representative aminopeptidases and prolyl endopeptidase from murine macrophages: comparative activity levels in resident and elicited cells. Biochem Pharmacol. 2005; 69: 1441–1450.
32.
BradfordMM.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72: 248–254.
33.
VaronaA, BlancoL, PerezI, . Expression and activity profiles of DPP IV/CD26 and NEP/CD10 glycoproteins in the human renal cancer are tumor-type dependent. BMC Cancer. 2010; 10: 193–204.
34.
LarrinagaG, PerezI, SanzB, . Angiotensin-converting enzymes (ACE and ACE2) are downregulated in renal tumors. Regul Peptide. 2010; 165: 218–223.
35.
CandenasL, SedaM, NohedaP, . Molecular diversity of voltage-gated sodium channel alpha and beta subunit mRNAs in human tissues. Eur J Pharmacol. 2006; 541: 9–16.
36.
VandesompeleJ, de PreterK, PattynF, . Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biology. 2002; 3:RESEARCH0034.
37.
GriffinJD, RitzJ, NadlerLM, . Expression of myeloid differentiation antigens on normal and malignant myeloid cells. J Clin Invest. 1981; 68: 932–941.
38.
QuaroniA, NicholsBL, QuaroniE, . Expression and different polarity of aminopeptidase N in normal human colonic mucosa and colonic tumors. Int J Cancer. 1992; 51 (3): 404–411.
39.
Birkenkamp-DemtroderK, ChristensenLL, OlesenSH, . Gene expression in colorectal cancer. Cancer Res. 2002; 62 (15): 4352–4363.
40.
MathieuMG, LinleyAJ, ReederSP, . HAGE, a cancer/testis antigen expressed at the protein level in a variety of cancers. Cancer Immun. 2010; 10: 2–9.
41.
PerezI, VaronaA, BlancoL, . Increased APN/CD13 and acid aminopeptidase activities in head and neck squamous cell carcinoma. Head Neck. 2009; 31 (10): 1335–1340.
42.
VaronaA, BlancoL, LópezJI, . Altered levels of acid, basic, and neutral peptidase activity and expression in human clear cell renal cell carcinoma. Am J Physiol-Renal Physiol. 2007; 292: 780–788.
43.
SchmittC, VoegelinM, MarinA, . Selective aminopeptidase-N (CD13) inhibitors with relevance to cancer chemotherapy. Bioorg Med Chem. 2013; 21 (7): 2135–2144.
44.
SantoroA, RimassaL, SobreroAF, . Phase II study of NGR-hTNF, a selective vascular targeting agent, in patients with metastatic colorectal cancer after failure of standard therapy. Eur J Cancer. 2010; 46 (15): 2746–2752.
