Top

Journal of Ovarian Research

Published in:

Open Access 01-12-2013 | Research

Investigation of human cationic antimicrobial protein-18 (hCAP-18), lactoferrin and CD163 as potential biomarkers for ovarian cancer

Authors: Ratana Lim, Martha Lappas, Clyde Riley, Niels Borregaard, Holger J Moller, Nuzhat Ahmed, Gregory E Rice

Published in: Journal of Ovarian Research | Issue 1/2013

Abstract

Background

Epithelial ovarian cancer is one of the leading causes of gynaecological cancer morbidity and mortality in women. Early stage ovarian cancer is usually asymptomatic, therefore, is often first diagnosed when it is widely disseminated. Currently available diagnostics lack the requisite sensitivity and specificity to be implemented as community-based screening tests. The identification of additional biomarkers may improve the diagnostic efficiency of multivariate index assays. The aims of this study were to characterise and compare the ovarian tissue immunohistochemical localisation and plasma concentrations of three putative ovarian cancer biomarkers: human cationic antimicrobial protein-18 (hCAP-18); lactoferrin; and CD163 in normal healthy women and women with ovarian cancer.

Methods

In this case–control cohort study, ovarian tissue and blood samples were obtained from 164 women (73 controls, including 28 women with benign pelvic masses; 91 cancer, including 21 women with borderline tumours). Localisation of each antigen within the ovary was assessed by immunohistochemistry and serum concentrations determined by ELISA assays.

Results

Immunoreactive (ir) hCAP-18 and lactoferrin were identified in epithelial cells, while CD163 was predominately localised in stromal cells. Tissue ir CD163 increased significantly (P<0.05) with disease grade. Median plasma concentrations of soluble (s)CD163 were significantly greater in the cases (3220 ng/ml) than in controls (2488 ng/ml) (P< 0.01). Median plasma concentrations of hCAP-18 and lactoferrin were not significantly different between cases and controls. The classification efficiency of each biomarker (as determined by the area under the receiver operator characteristic curve; AUC) was: 0.67± 0.04; 0.62 ± 0.08 and 0.51 ± 0.07 for sCD163, hCAP-18 and lactoferrin, respectively. When the 3 biomarkers were modelled using stochastic gradient boosted logistic regression, the AUC increased to 0.95 ± 0.03.

Conclusions

The data obtained in this study establishes the localisation and concentrations of CD163, hCAP-18, and lactoferrin in ovarian tumours and peripheral blood. Individually, the 3 biomarkers display only modest diagnostic efficiency as assessed by AUC. When combined in a multivariate index assay, however, diagnostic efficiency increases significantly. As such, the utility of the biomarker panel, as an aid in the diagnosis of cancer in symptomatic women, is worthy of further investigation in a larger phase 2 biomarker trial.

Available only for authorised users

Sankaranarayanan R, Ferlay J: Worldwide burden of gynaecological cancer: the size of the problem. Best Pract Res Clin Obstet Gynaecol 2006, 20: 207–225. 10.1016/j.bpobgyn.2005.10.007PubMedCrossRef

Howlader N, Noone AM, Krapcho M, Neyman N, Aminou R, Altekruse SF, Kosary CL, Ruhl J, Tatalovich Z, Cho H, Mariotto A, Eisner M, et al.: SEER Cancer Statistics Review, 1975–2009 (Vintage 2009 Populations). Bethesda, MD: National Cancer Institute; 2012. http://seer.cancer.gov/csr/1975_2009_pops09

Paley PJ: Ovarian cancer screening: are we making any progress? Curr Opin Oncol 2001, 13: 399–402. 10.1097/00001622-200109000-00015PubMedCrossRef

Lenhard M, Stieber P, Hertlein L, Kirschenhofer A, Furst S, Mayr D, Nagel D, Hofmann K, Krocker K, Burges A: The diagnostic accuracy of two human epididymis protein 4 (HE4) testing systems in combination with CA125 in the differential diagnosis of ovarian masses. Clin Chem Lab Med 2011, 49: 2081–2088.PubMedCrossRef

Bjorge T, Lukanova A, Tretli S, Manjer J, Ulmer H, Stocks T, Selmer R, Nagel G, Almquist M, Concin H, Hallmans G, Jonsson H, et al.: Metabolic risk factors and ovarian cancer in the metabolic syndrome and cancer project. Int J Epidemiol 2011, 40: 1667–1677. 10.1093/ije/dyr130PubMedCrossRef

Sorensen O, Cowland JB, Askaa J, Borregaard N: An ELISA for hCAP-18, the cathelicidin present in human neutrophils and plasma. J Immunol Methods 1997, 206: 53–59. 10.1016/S0022-1759(97)00084-7PubMedCrossRef

Altman H, Steinberg D, Porat Y, Mor A, Fridman D, Friedman M, Bachrach G: In vitro assessment of antimicrobial peptides as potential agents against several oral bacteria. J Antimicrob Chemother 2006, 58: 198–201. 10.1093/jac/dkl181PubMedCrossRef

Huang LC, Petkova TD, Reins RY, Proske RJ, McDermott AM: Multifunctional roles of human cathelicidin (LL-37) at the ocular surface. Invest Ophthalmol Vis Sci 2006, 47: 2369–2380. 10.1167/iovs.05-1649PubMedCrossRef

Frohm Nilsson M, Sandstedt B, Sorensen O, Weber G, Borregaard N, Stahle-Backdahl M: The human cationic antimicrobial protein (hCAP18), a peptide antibiotic, is widely expressed in human squamous epithelia and colocalizes with interleukin-6. Infect Immun 1999, 67: 2561–2566.PubMedCentralPubMed

10.

Frohm M, Agerberth B, Ahangari G, Stahle-Backdahl M, Liden S, Wigzell H, Gudmundsson GH: The expression of the gene coding for the antibacterial peptide LL-37 is induced in human keratinocytes during inflammatory disorders. J Biol Chem 1997, 272: 15258–15263. 10.1074/jbc.272.24.15258PubMedCrossRef

11.

Coffelt SB, Waterman RS, Florez L, zu Bentrup KH, Zwezdaryk KJ, Tomchuck SL, LaMarca HL, Danka ES, Morris CA, Scandurro AB: Ovarian cancers overexpress the antimicrobial protein hCAP-18 and its derivative LL-37 increases ovarian cancer cell proliferation and invasion. Int J Cancer 2008, 122: 1030–1039.PubMedCrossRef

12.

Groves ML: The isolation of a Red protein from milk. J Am Chem Soc 1960, 82: 3345–3350. 10.1021/ja01498a029CrossRef

13.

Johanson B: Isolation of an iron-containing red protein from human milk. Acta Chemica Scandinavica 1960, 14: 510–512.CrossRef

14.

Montreuil J, Tonnelat J, Mullet S: Preparation and properties of lactosiderophilin (lactotransferrin) of human milk. Biochim Biophys Acta 1960, 45: 413–421.PubMedCrossRef

15.

Brock JH: Lactoferrin in human milk: its role in iron absorption and protection against enteric infection in the newborn infant. Arch Dis Child 1980, 55: 417–421. 10.1136/adc.55.6.417PubMedCentralPubMedCrossRef

16.

Lonnerdal B, Forsum E, Gebre-Medhin M, Hambraeus L: Breast milk composition in Ethiopian and Swedish mothers. II. Lactose, nitrogen, and protein contents. Am J Clin Nutr 1976, 29: 1134–1141.PubMed

17.

Levay PF, Viljoen M: Lactoferrin: a general review. Haematologica 1995, 80: 252–267.PubMed

18.

Baggiolini M, Hirsch JG, De Duve C: Further biochemical and morphological studies of granule fractions from rabbit heterophil leukocytes. J Cell Biol 1970, 45: 586–597. 10.1083/jcb.45.3.586PubMedCentralPubMedCrossRef

19.

Masson PL, Heremans JF, Schonne E: Lactoferrin, an iron-binding protein in neutrophilic leukocytes. J Exp Med 1969, 130: 643–658. 10.1084/jem.130.3.643PubMedCentralPubMedCrossRef

20.

Ward PP, Uribe-Luna S, Conneely OM: Lactoferrin and host defense. Biochem Cell Biol 2002, 80: 95–102. 10.1139/o01-214PubMedCrossRef

21.

Kholodnyuk ID, Kozireva S, Kost-Alimova M, Kashuba V, Klein G, Imreh S: Down regulation of 3p genes, LTF, SLC38A3 and DRR1, upon growth of human chromosome 3-mouse fibrosarcoma hybrids in severe combined immunodeficiency mice. Int J Cancer 2006, 119: 99–107. 10.1002/ijc.21794PubMedCrossRef

22.

Newbold RR, Jefferson WN, Grissom SF, Padilla-Banks E, Snyder RJ, Lobenhofer EK: Developmental exposure to diethylstilbestrol alters uterine gene expression that may be associated with uterine neoplasia later in life. Mol Carcinog 2007, 46: 783–796. 10.1002/mc.20308PubMedCentralPubMedCrossRef

23.

Cao L, Zhou Y, Li X, Yi H: The relationship of haplotype in lactotransferrin and its expression levels in chinese Han ovarian cancer. Acta Biochim Biophys Sin (Shanghai) 2011, 43: 884–890. 10.1093/abbs/gmr089CrossRef

24.

Moestrup SK, Moller HJ: CD163: a regulated hemoglobin scavenger receptor with a role in the anti-inflammatory response. Ann Med 2004, 36: 347–354. 10.1080/07853890410033171PubMedCrossRef

25.

Moller HJ, Aerts H, Gronbaek H, Peterslund NA, Hyltoft Petersen P, Hornung N, Rejnmark L, Jabbarpour E, Moestrup SK: Soluble CD163: a marker molecule for Monocyte/macrophage activity in disease. Scand J Clin Lab Invest Suppl 2002, 237: 29–33.PubMedCrossRef

26.

Greisen SR, Moller HJ, Stengaard-Pedersen K, Hetland ML, Horslev-Petersen K, Jorgensen A, Hvid M, Deleuran B: Soluble macrophage-derived CD163 is a marker of disease activity and progression in early rheumatoid arthritis. Clin Exp Rheumatol 2011, 29: 689–692.PubMed

27.

Matsushita N, Kashiwagi M, Wait R, Nagayoshi R, Nakamura M, Matsuda T, Hogger P, Guyre PM, Nagase H, Matsuyama T: Elevated levels of soluble CD163 in sera and fluids from rheumatoid arthritis patients and inhibition of the shedding of CD163 by TIMP-3. Clin Exp Immunol 2002, 130: 156–161. 10.1046/j.1365-2249.2002.01963.xPubMedCentralPubMedCrossRef

28.

Knudsen TB, Gustafson P, Kronborg G, Kristiansen TB, Moestrup SK, Nielsen JO, Gomes V, Aaby P, Lisse I, Moller HJ, Eugen-Olsen J: Predictive value of soluble haemoglobin scavenger receptor CD163 serum levels for survival in verified tuberculosis patients. Clin Microbiol Infect 2005, 11: 730–735. 10.1111/j.1469-0691.2005.01229.xPubMedCrossRef

29.

Kawamura K, Komohara Y, Takaishi K, Katabuchi H, Takeya M: Detection of M2 macrophages and colony-stimulating factor 1 expression in serous and mucinous ovarian epithelial tumors. Pathol Int 2009, 59: 300–305. 10.1111/j.1440-1827.2009.02369.xPubMedCrossRef

30.

Wang R, Zhang T, Ma Z, Wang Y, Cheng Z, Xu H, Li W, Wang X: The interaction of coagulation factor XII and monocyte/macrophages mediating peritoneal metastasis of epithelial ovarian cancer. Gynecol Oncol 2010, 117: 460–466. 10.1016/j.ygyno.2010.02.015PubMedCrossRef

31.

Zhang T, Ma Z, Wang R, Wang Y, Wang S, Cheng Z, Xu H, Jin X, Li W, Wang X: Thrombin facilitates invasion of ovarian cancer along peritoneum by inducing monocyte differentiation toward tumor-associated macrophage-like cells. Cancer Immunol Immunother 2010, 59: 1097–1108. 10.1007/s00262-010-0836-yPubMedCrossRef

32.

Pepe MS: Evaluating technologies for classification and prediction in medicine. Stat Med 2005, 19: 3687–3696.CrossRef

33.

Silverberg SG: Histopathologic grading of ovarian carcinoma: a review and proposal. Int J Gynecol Pathol 2000, 19: 7–15. 10.1097/00004347-200001000-00003PubMedCrossRef

34.

Ahmed N, Barker G, Oliva KT, Hoffmann P, Riley C, Reeve S, Smith AI, Kemp BE, Quinn MA, Rice GE: Proteomic-based identification of haptoglobin-1 precursor as a novel circulating biomarker of ovarian cancer. Br J Cancer 2004, 91: 129–140. 10.1038/sj.bjc.6601882PubMedCentralPubMedCrossRef

35.

Lim R, Ahmed N, Borregaard N, Riley C, Wafai R, Thompson EW, Quinn MA, Rice GE: Neutrophil gelatinase-associated lipocalin (NGAL) an early-screening biomarker for ovarian cancer: NGAL is associated with epidermal growth factor-induced epithelio-mesenchymal transition. Int J Cancer 2007, 120: 2426–2434. 10.1002/ijc.22352PubMedCrossRef

36.

Armes JE, Trute L, White D, Southey MC, Hammet F, Tesoriero A, Hutchins AM, Dite GS, McCredie MR, Giles GG, Hopper JL, Venter DJ: Distinct molecular pathogeneses of early-onset breast cancers in BRCA1 and BRCA2 mutation carriers: a population-based study. Cancer Res 1999, 59: 2011–2017.PubMed

37.

Birgens HS: Lactoferrin in plasma measured by an ELISA technique: evidence that plasma lactoferrin is an indicator of neutrophil turnover and bone marrow activity in acute leukaemia. Scand J Haematol 1985, 34: 326–331.PubMedCrossRef

38.

Moller HJ, Hald K, Moestrup SK: Characterization of an enzyme-linked immunosorbent assay for soluble CD163. Scand J Clin Lab Invest 2002, 62: 293–299. 10.1080/003655102760145852PubMedCrossRef

39.

Waegeman W, De Baets B, Boullart L: Learning layered ranking functions with structured support vector machines. Neural Netw 2008, 21: 1511–1523. 10.1016/j.neunet.2008.07.008PubMedCrossRef

40.

Hanley JA, McNeil BJ: A method of comparing the areas under receiver operating characteristic curves derived from the same cases. Radiology 1983, 148: 839–843.PubMedCrossRef

41.

Friedman N, Linial M, Nachman I, Pe'er D: Using bayesian networks to analyze expression data. J Comput Biol 2000, 7: 601–620. 10.1089/106652700750050961PubMedCrossRef

42.

Hall M, Frank E, Holmes G, Pfahringer B, Reutemann P, Witten IH: The WEKA data mining software: an update. SIGKDD Explor. Newsl. 2009, 11: 10–18. 10.1145/1656274.1656278CrossRef

43.

Heilborn JD, Nilsson MF, Jimenez CI, Sandstedt B, Borregaard N, Tham E, Sorensen OE, Weber G, Stahle M: Antimicrobial protein hCAP18/LL-37 is highly expressed in breast cancer and is a putative growth factor for epithelial cells. Int J Cancer 2005, 114: 713–719. 10.1002/ijc.20795PubMedCrossRef

44.

Weber G, Chamorro CI, Granath F, Liljegren A, Zreika S, Saidak Z, Sandstedt B, Rotstein S, Mentaverri R, Sanchez F, Pivarcsi A, Stahle M: Human antimicrobial protein hCAP18/LL-37 promotes a metastatic phenotype in breast cancer. Breast Cancer Res 2009, 11: R6. 10.1186/bcr2221PubMedCentralPubMedCrossRef

45.

von Haussen J, Koczulla R, Shaykhiev R, Herr C, Pinkenburg O, Reimer D, Wiewrodt R, Biesterfeld S, Aigner A, Czubayko F, Bals R: The host defence peptide LL-37/hCAP-18 is a growth factor for lung cancer cells. Lung Cancer 2008, 59: 12–23. 10.1016/j.lungcan.2007.07.014PubMedCrossRef

46.

Sorensen OE, Cowland JB, Theilgaard-Monch K, Liu L, Ganz T, Borregaard N: Wound healing and expression of antimicrobial peptides/polypeptides in human keratinocytes, a consequence of common growth factors. J Immunol 2003, 170: 5583–5589.PubMedCrossRef

47.

Tjabringa GS, Rabe KF, Hiemstra PS: The human cathelicidin LL-37: a multifunctional peptide involved in infection and inflammation in the lung. Pulm Pharmacol Ther 2005, 18: 321–327. 10.1016/j.pupt.2005.01.001PubMedCrossRef

48.

Nguyen TT, Schwartz EJ, West RB, Warnke RA, Arber DA, Natkunam Y: Expression of CD163 (hemoglobin scavenger receptor) in normal tissues, lymphomas, carcinomas, and sarcomas is largely restricted to the monocyte/macrophage lineage. Am J Surg Pathol 2005, 29: 617–624. 10.1097/01.pas.0000157940.80538.ecPubMedCrossRef

49.

Jensen TO, Schmidt H, Moller HJ, Hoyer M, Maniecki MB, Sjoegren P, Christensen IJ, Steiniche T: Macrophage markers in serum and tumor have prognostic impact in american joint committee on cancer stage I/II melanoma. J Clin Oncol 2009, 27: 3330–3337. 10.1200/JCO.2008.19.9919PubMedCrossRef

50.

Maniecki MB, Etzerodt A, Ulhoi BP, Steiniche T, Borre M, Dyrskjot L, Orntoft TF, Moestrup SK, Moller HJ: Tumor-promoting macrophages induce the expression of the macrophage-specific receptor CD163 in malignant cells. Int J Cancer 2012, 131: 2320–2331. 10.1002/ijc.27506PubMedCrossRef

51.

Allavena P, Mantovani A: Immunology in the clinic review series; focus on cancer: tumour-associated macrophages: undisputed stars of the inflammatory tumour microenvironment. Clin Exp Immunol 2012, 167: 195–205. 10.1111/j.1365-2249.2011.04515.xPubMedCentralPubMedCrossRef

52.

Van Gorp H, Delputte PL, Nauwynck HJ: Scavenger receptor CD163, a jack-of-all-trades and potential target for cell-directed therapy. Mol Immunol 2010, 47: 1650–1660. 10.1016/j.molimm.2010.02.008PubMedCrossRef

53.

Lee CH, Espinosa I, Vrijaldenhoven S, Subramanian S, Montgomery KD, Zhu S, Marinelli RJ, Peterse JL, Poulin N, Nielsen TO, West RB, Gilks CB, et al.: Prognostic significance of macrophage infiltration in leiomyosarcomas. Clin Cancer Res 2008, 14: 1423–1430. 10.1158/1078-0432.CCR-07-1712PubMedCrossRef

54.

Moller HJ: Soluble CD163. Scand J Clin Lab Invest 2012, 72: 1–13. 10.3109/00365513.2011.626868PubMedCrossRef

Title: Investigation of human cationic antimicrobial protein-18 (hCAP-18), lactoferrin and CD163 as potential biomarkers for ovarian cancer
Authors: Ratana Lim
Martha Lappas
Clyde Riley
Niels Borregaard
Holger J Moller
Nuzhat Ahmed
Gregory E Rice
Publication date: 01-12-2013
Publisher: BioMed Central
Published in: Journal of Ovarian Research / Issue 1/2013
Electronic ISSN: 1757-2215
DOI: https://doi.org/10.1186/1757-2215-6-5

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Investigation of human cationic antimicrobial protein-18 (hCAP-18), lactoferrin and CD163 as potential biomarkers for ovarian cancer

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2013

Safe and targeted anticancer therapy for ovarian cancer using a novel class of curcumin analogs

Immature ovarian teratoma with unusual gliomatosis

GPER-1 acts as a tumor suppressor in ovarian cancer

Clinical governance network for clinical audit to improve quality in epithelial ovarian cancer management

The significance of the alteration of 8-OHdG in serous ovarian carcinoma

Ovarian masses in children and adolescents in China: analysis of 203 cases