Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
Journal of Ovarian Research
Published in: Journal of Ovarian Research 1/2010

Open Access 01-12-2010 | Research

Leucine-rich alpha-2-glycoprotein-1 is upregulated in sera and tumors of ovarian cancer patients

Authors: John D Andersen, Kristin LM Boylan, Ronald Jemmerson, Melissa A Geller, Benjamin Misemer, Katherine M Harrington, Starchild Weivoda, Bruce A Witthuhn, Peter Argenta, Rachel Isaksson Vogel, Amy PN Skubitz

Published in: Journal of Ovarian Research | Issue 1/2010

Login to get access

Abstract

Background

New biomarkers that replace or are used in conjunction with the current ovarian cancer diagnostic antigen, CA125, are needed for detection of ovarian cancer in the presurgical setting, as well as for detection of disease recurrence. We previously demonstrated the upregulation of leucine-rich alpha-2-glycoprotein-1 (LRG1) in the sera of ovarian cancer patients compared to healthy women using quantitative mass spectrometry.

Methods

LRG1 was quantified by ELISA in serum from two relatively large cohorts of women with ovarian cancer and benign gynecological disease. The expression of LRG1 in ovarian cancer tissues and cell lines was examined by gene microarray, reverse-transcriptase polymerase chain reaction (RT-PCR), Western blot, immunocytochemistry and mass spectrometry.

Results

Mean serum LRG1 was higher in 58 ovarian cancer patients than in 56 healthy women (89.33 ± 77.90 vs. 42.99 ± 9.88 ug/ml; p = 0.0008) and was highest among stage III/IV patients. In a separate set of 193 pre-surgical samples, LRG1 was higher in patients with serous or clear cell ovarian cancer (145.82 ± 65.99 ug/ml) compared to patients with benign gynecological diseases (82.53 ± 76.67 ug/ml, p < 0.0001). CA125 and LRG1 levels were moderately correlated (r = 0.47, p < 0.0001). LRG1 mRNA levels were higher in ovarian cancer tissues and cell lines compared to their normal counterparts when analyzed by gene microarray and RT-PCR. LRG1 protein was detected in ovarian cancer tissue samples and cell lines by immunocytochemistry and Western blotting. Multiple iosforms of LRG1 were observed by Western blot and were shown to represent different glycosylation states by digestion with glycosidase. LRG1 protein was also detected in the conditioned media of ovarian cancer cell culture by ELISA, Western blotting, and mass spectrometry.

Conclusions

Serum LRG1 was significantly elevated in women with ovarian cancer compared to healthy women and women with benign gynecological disease, and was only moderately correlated with CA125. Ovarian cancer cells secrete LRG1 and may contribute directly to the elevated levels of LRG1 observed in the serum of ovarian cancer patients. Future studies will determine whether LRG1 may serve as a biomarker for presurgical diagnosis, disease recurrence, and/or as a target for therapy.
Appendix
Available only for authorised users
Literature
1.
Ries L, Harkins D, Krapcho M, Mariotto A, Miller BA, Feuer EJ, Clegg L, Eisner MP, Horner MJ, Howlader N, Hayat M, Hankey BF, Edwards BK: SEER Cancer Statistics Review, 1975–2003. Book SEER Cancer Statistics Review, 1975–2003 (Editor ed.^eds.). City 2005.
2.
Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, Thun MJ: Cancer statistics, 2008. CA Cancer J Clin 2008, 58: 71–96. 10.3322/CA.2007.0010CrossRefPubMed
3.
4.
Yurkovetsky ZR, Linkov FY, D EM, Lokshin AE: Multiple biomarker panels for early detection of ovarian cancer. Future Oncol 2006, 2: 733–741. 10.2217/14796694.2.6.733CrossRefPubMed
5.
Fung ET: A recipe for proteomics diagnostic test development: the OVA1 test, from biomarker discovery to FDA clearance. Clin Chem 56: 327–329. 10.1373/clinchem.2009.140855
6.
Mercado C, Zingmond D, Karlan BY, Sekaris E, Gross J, Maggard-Gibbons M, Tomlinson JS, Ko CY: Quality of care in advanced ovarian cancer: the importance of provider specialty. Gynecol Oncol 117: 18–22. 10.1016/j.ygyno.2009.12.033
7.
Anderson NL, Anderson NG: The human plasma proteome: history, character, and diagnostic prospects. Mol Cell Proteomics 2002, 1: 845–867. 10.1074/mcp.R200007-MCP200CrossRefPubMed
8.
Andersen JD, Boylan KLM, Xue FS, Anderson LB, Witthuhn BA, Markowski TW, Higgins L, Skubitz APN: Identification of candidate biomarkers in ovarian cancer serum by depletion of highly abundant proteins and differential in-gel electrophoresis. Electrophoresis 2010., 31: 10.1002/elps.200900441
9.
Boylan KLM, Andersen JD, Anderson LB, Higgins L, Skubitz APN: Quantitative proteomic analysis by iTRAQ®for the identification of candidate biomarkers in ovarian cancer serum. Proteome Sci 2010, 8: 31. 10.1186/1477-5956-8-31PubMedCentralCrossRefPubMed
10.
Schwick HG, Haupt H: Purified human plasma proteins of unknown function. Jpn J Med Sci Biol 1981, 34: 299–327.CrossRefPubMed
11.
Hoogland C, Mostaguir K, Sanchez JC, Hochstrasser DF, Appel RD: SWISS-2DPAGE, ten years later. Proteomics 2004, 4: 2352–2356. 10.1002/pmic.200300830CrossRefPubMed
12.
Haupt H, Baudner S: [Isolation and characterization of an unknown, leucine-rich 3.1-S-alpha2-glycoprotein from human serum (author's transl)]. Hoppe Seylers Z Physiol Chem 1977, 358: 639–646.CrossRefPubMed
13.
Weivoda S, Andersen JD, Skogen A, Schlievert PM, Fontana D, Schacker T, Tuite P, Dubinsky JM, Jemmerson R: ELISA for human serum leucine-rich alpha-2-glycoprotein-1 employing cytochrome c as the capturing ligand. J Immunol Methods 2008, 336: 22–29. 10.1016/j.jim.2008.03.004CrossRefPubMed
14.
Kobe B, Kajava AV: The leucine-rich repeat as a protein recognition motif. Curr Opin Struct Biol 2001, 11: 725–732. 10.1016/S0959-440X(01)00266-4CrossRefPubMed
15.
Takahashi N, Takahashi Y, Putnam FW: Periodicity of leucine and tandem repetition of a 24-amino acid segment in the primary structure of leucine-rich alpha 2-glycoprotein of human serum. Proc Natl Acad Sci USA 1985, 82: 1906–1910. 10.1073/pnas.82.7.1906PubMedCentralCrossRefPubMed
16.
O'Donnell LC, Druhan LJ, Avalos BR: Molecular characterization and expression analysis of leucine-rich alpha2-glycoprotein, a novel marker of granulocytic differentiation. J Leukoc Biol 2002, 72: 478–485.PubMed
17.
Saito K, Tanaka T, Kanda H, Ebisuno Y, Izawa D, Kawamoto S, Okubo K, Miyasaka M: Gene expression profiling of mucosal addressin cell adhesion molecule-1+ high endothelial venule cells (HEV) and identification of a leucine-rich HEV glycoprotein as a HEV marker. J Immunol 2002, 168: 1050–1059.CrossRefPubMed
18.
Sun D, Kar S, Carr BI: Differentially expressed genes in TGF-beta 1 sensitive and resistant human hepatoma cells. Cancer Lett 1995, 89: 73–79.CrossRefPubMed
19.
Li X, Miyajima M, Jiang C, Arai H: Expression of TGF-betas and TGF-beta type II receptor in cerebrospinal fluid of patients with idiopathic normal pressure hydrocephalus. Neurosci Lett 2007, 413: 141–144. 10.1016/j.neulet.2006.11.039CrossRefPubMed
20.
Cummings C, Walder J, Treeful A, Jemmerson R: Serum leucine-rich alpha-2-glycoprotein-1 binds cytochrome c and inhibits antibody detection of this apoptotic marker in enzyme-linked immunosorbent assay. Apoptosis 2006, 11: 1121–1129. 10.1007/s10495-006-8159-3CrossRefPubMed
21.
Ai J, Druhan LJ, Hunter MG, Loveland MJ, Avalos BR: LRG-accelerated differentiation defines unique G-CSFR signaling pathways downstream of PU.1 and C/EBPepsilon that modulate neutrophil activation. J Leukoc Biol 2008, 83: 1277–1285. 10.1189/jlb.1107751PubMedCentralCrossRefPubMed
22.
Hibbs K, Skubitz KM, Pambuccian SE, Casey RC, Burleson KM, Oegema TR Jr, Thiele JJ, Grindle SM, Bliss RL, Skubitz AP: Differential gene expression in ovarian carcinoma: identification of potential biomarkers. Am J Pathol 2004, 165: 397–414.PubMedCentralCrossRefPubMed
23.
Skubitz AP, Pambuccian SE, Argenta PA, Skubitz KM: Differential gene expression identifies subgroups of ovarian carcinoma. Transl Res 2006, 148: 223–248. 10.1016/j.trsl.2006.06.001CrossRefPubMed
24.
DeRycke MS, Andersen JD, Harrington KM, Pambuccian SE, Kalloger SE, Boylan KL, Argenta PA, Skubitz AP: S100A1 expression in ovarian and endometrial endometrioid carcinomas is a prognostic indicator of relapse-free survival. Am J Clin Pathol 2009, 132: 846–856. 10.1309/AJCPTK87EMMIKPFSPubMedCentralCrossRefPubMed
25.
Subramanian IV, Bui Nguyen TM, Truskinovsky AM, Tolar J, Blazar BR, Ramakrishnan S: Adeno-associated virus-mediated delivery of a mutant endostatin in combination with carboplatin treatment inhibits orthotopic growth of ovarian cancer and improves long-term survival. Cancer Res 2006, 66: 4319–4328. 10.1158/0008-5472.CAN-05-3297CrossRefPubMed
26.
Shaw TJ, Senterman MK, Dawson K, Crane CA, Vanderhyden BC: Characterization of intraperitoneal, orthotopic, and metastatic xenograft models of human ovarian cancer. Mol Ther 2004, 10: 1032–1042. 10.1016/j.ymthe.2004.08.013CrossRefPubMed
27.
Kruk PA, Maines-Bandiera SL, Auersperg N: A simplified method to culture human ovarian surface epithelium. Lab Invest 1990, 63: 132–136.PubMed
28.
Nicosia SV, Wilbanks GD, Saunders B, Mayer J, Cardosi RJ, Kruk PA, Cheng J, Bai W, Coppola D, Fiorica J: Cytology of human ovarian surface epithelial brushings. Cancer 2004, 102: 1–10. 10.1002/cncr.20001CrossRefPubMed
29.
Shirai R, Hirano F, Ohkura N, Ikeda K, Inoue S: Up-regulation of the expression of leucine-rich alpha(2)-glycoprotein in hepatocytes by the mediators of acute-phase response. Biochem Biophys Res Commun 2009, 382: 776–779. 10.1016/j.bbrc.2009.03.104CrossRefPubMed
30.
Keller A, Nesvizhskii AI, Kolker E, Aebersold R: Empirical statistical model to estimate the accuracy of peptide identifications made by MS/MS and database search. Anal Chem 2002, 74: 5383–5392. 10.1021/ac025747hCrossRefPubMed
31.
Govorukhina NI, de Vries M, Reijmers TH, Horvatovich P, van der Zee AG, Bischoff R: Influence of clotting time on the protein composition of serum samples based on LC-MS data. J Chromatogr B Analyt Technol Biomed Life Sci 2009, 877: 1281–1291. 10.1016/j.jchromb.2008.10.029CrossRefPubMed
32.
Bini L, Magi B, Marzocchi B, Cellesi C, Berti B, Raggiaschi R, Rossolini A, Pallini V: Two-dimensional electrophoretic patterns of acute-phase human serum proteins in the course of bacterial and viral diseases. Electrophoresis 1996, 17: 612–616. 10.1002/elps.1150170333CrossRefPubMed
33.
Norkina O, Kaur S, Ziemer D, De Lisle RC: Inflammation of the cystic fibrosis mouse small intestine. Am J Physiol Gastrointest Liver Physiol 2004, 286: G1032–1041. 10.1152/ajpgi.00473.2003CrossRefPubMed
34.
Kawakami T, Hoshida Y, Kanai F, Tanaka Y, Tateishi K, Ikenoue T, Obi S, Sato S, Teratani T, Shiina S, et al.: Proteomic analysis of sera from hepatocellular carcinoma patients after radiofrequency ablation treatment. Proteomics 2005, 5: 4287–4295. 10.1002/pmic.200401287CrossRefPubMed
35.
Yu KH, Rustgi AK, Blair IA: Characterization of proteins in human pancreatic cancer serum using differential gel electrophoresis and tandem mass spectrometry. J Proteome Res 2005, 4: 1742–1751. 10.1021/pr050174lCrossRefPubMed
36.
Kakisaka T, Kondo T, Okano T, Fujii K, Honda K, Endo M, Tsuchida A, Aoki T, Itoi T, Moriyasu F, et al.: Plasma proteomics of pancreatic cancer patients by multi-dimensional liquid chromatography and two-dimensional difference gel electrophoresis (2D-DIGE): up-regulation of leucine-rich alpha-2-glycoprotein in pancreatic cancer. J Chromatogr B Analyt Technol Biomed Life Sci 2007, 852: 257–267. 10.1016/j.jchromb.2007.01.029CrossRefPubMed
37.
Okano T, Kondo T, Kakisaka T, Fujii K, Yamada M, Kato H, Nishimura T, Gemma A, Kudoh S, Hirohashi S: Plasma proteomics of lung cancer by a linkage of multi-dimensional liquid chromatography and two-dimensional difference gel electrophoresis. Proteomics 2006, 6: 3938–3948. 10.1002/pmic.200500883CrossRefPubMed
38.
Zeng JW, Wang SX, Zhao XF, Wei MT, Tu ZG: [Utilizing 2-DE and MALDI-TOF MS/MS to screen differentially expressed serum proteins of silicosis]. Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi 2007, 25: 136–141.PubMed
39.
Faca VM, Ventura AP, Fitzgibbon MP, Pereira-Faca SR, Pitteri SJ, Green AE, Ireton RC, Zhang Q, Wang H, O'Briant KC, et al.: Proteomic analysis of ovarian cancer cells reveals dynamic processes of protein secretion and shedding of extra-cellular domains. PLoS ONE 2008, 3: e2425. 10.1371/journal.pone.0002425PubMedCentralCrossRefPubMed
40.
Gortzak-Uzan L, Ignatchenko A, Evangelou AI, Agochiya M, Brown KA, St Onge P, Kireeva I, Schmitt-Ulms G, Brown TJ, Murphy J, et al.: A proteome resource of ovarian cancer ascites: integrated proteomic and bioinformatic analyses to identify putative biomarkers. J Proteome Res 2008, 7: 339–351. 10.1021/pr0703223CrossRefPubMed
41.
Pan S, Cheng L, White JT, Lu W, Utleg AG, Yan X, Urban ND, Drescher CW, Hood L, Lin B: Quantitative proteomics analysis integrated with microarray data reveals that extracellular matrix proteins, catenins, and p53 binding protein 1 are important for chemotherapy response in ovarian cancers. Omics 2009, 13: 345–354. 10.1089/omi.2009.0008CrossRefPubMed
42.
Lin B, White JT, Wu J, Lele S, Old LJ, Hood L, Odunsi K: Deep depletion of abundant serum proteins reveals low-abundant proteins as potential biomarkers for human ovarian cancer. Proteomics Clin Appl 2009, 3: 853–861. 10.1002/prca.200800141PubMedCentralCrossRefPubMed
43.
Sardana G, Jung K, Stephan C, Diamandis EP: Proteomic analysis of conditioned media from the PC3, LNCaP, and 22Rv1 prostate cancer cell lines: discovery and validation of candidate prostate cancer biomarkers. J Proteome Res 2008, 7: 3329–3338. 10.1021/pr8003216CrossRefPubMed
44.
Sardana G, Marshall J, Diamandis EP: Discovery of candidate tumor markers for prostate cancer via proteomic analysis of cell culture-conditioned medium. Clin Chem 2007, 53: 429–437. 10.1373/clinchem.2006.077370CrossRefPubMed
45.
Kulasingam V, Diamandis EP: Tissue culture-based breast cancer biomarker discovery platform. Int J Cancer 2008, 123: 2007–2012. 10.1002/ijc.23844CrossRefPubMed
46.
Ferrero S, Gillott DJ, Remorgida V, Anserini P, Ragni N, Grudzinskas JG: Increased expression of one isoform of leucine-rich alpha-2-glycoprotein in peritoneal fluid of women with uterine leiomyomas. Arch Gynecol Obstet 2009, 279: 365–371. 10.1007/s00404-008-0741-1CrossRefPubMed
47.
Ye B, Cramer DW, Skates SJ, Gygi SP, Pratomo V, Fu L, Horick NK, Licklider LJ, Schorge JO, Berkowitz RS, Mok SC: Haptoglobin-alpha subunit as potential serum biomarker in ovarian cancer: identification and characterization using proteomic profiling and mass spectrometry. Clin Cancer Res 2003, 9: 2904–2911.PubMed
48.
Hirokawa T, Boon-Chieng S, Mitaku S: SOSUI: classification and secondary structure prediction system for membrane proteins. Bioinformatics 1998, 14: 378–379. 10.1093/bioinformatics/14.4.378CrossRefPubMed
49.
An HJ, Lebrilla CB: A glycomics approach to the discovery of potential cancer biomarkers. Methods Mol Biol 600: 199–213. full_text
50.
Saldova R, Royle L, Radcliffe CM, Abd Hamid UM, Evans R, Arnold JN, Banks RE, Hutson R, Harvey DJ, Antrobus R, et al.: Ovarian cancer is associated with changes in glycosylation in both acute-phase proteins and IgG. Glycobiology 2007, 17: 1344–1356. 10.1093/glycob/cwm100CrossRefPubMed
51.
Saldova R, Wormald MR, Dwek RA, Rudd PM: Glycosylation changes on serum glycoproteins in ovarian cancer may contribute to disease pathogenesis. Dis Markers 2008, 25: 219–232.PubMedCentralCrossRefPubMed
52.
Fujimura T, Shinohara Y, Tissot B, Pang PC, Kurogochi M, Saito S, Arai Y, Sadilek M, Murayama K, Dell A, et al.: Glycosylation status of haptoglobin in sera of patients with prostate cancer vs. benign prostate disease or normal subjects. Int J Cancer 2008, 122: 39–49. 10.1002/ijc.22958CrossRefPubMed
53.
Casey RC, Oegema TR Jr, Skubitz KM, Pambuccian SE, Grindle SM, Skubitz AP: Cell membrane glycosylation mediates the adhesion, migration, and invasion of ovarian carcinoma cells. Clin Exp Metastasis 2003, 20: 143–152. 10.1023/A:1022670501667CrossRefPubMed
Metadata
Title
Leucine-rich alpha-2-glycoprotein-1 is upregulated in sera and tumors of ovarian cancer patients
Authors
John D Andersen
Kristin LM Boylan
Ronald Jemmerson
Melissa A Geller
Benjamin Misemer
Katherine M Harrington
Starchild Weivoda
Bruce A Witthuhn
Peter Argenta
Rachel Isaksson Vogel
Amy PN Skubitz
Publication date
01-12-2010
Publisher
BioMed Central
Published in
Journal of Ovarian Research / Issue 1/2010
Electronic ISSN: 1757-2215
DOI
https://doi.org/10.1186/1757-2215-3-21

Other articles of this Issue 1/2010

Journal of Ovarian Research 1/2010 Go to the issue

Research

Augmented inhibition of angiogenesis by combination of HER2 antibody chA21 and trastuzumab in human ovarian carcinoma xenograft

Research

Expression of interleukin-1 (IL-1) ligands system in the most common endometriosis-associated ovarian cancer subtypes

Research

Curcumin induces chemo/radio-sensitization in ovarian cancer cells and curcumin nanoparticles inhibit ovarian cancer cell growth

Research

Targeting of mesenchymal stem cells to ovarian tumors via an artificial receptor

Case report

Struma ovarii associated with pseudo-Meigs' syndrome and elevated serum CA 125: a case report and review of the literature

Research

The prosurvival activity of ascites against TRAIL is associated with a shorter disease-free interval in patients with ovarian cancer