Prostate-specific antigen (PSA) has been widely used as the unique serum biomarker for the diagnosis of prostate cancer (PCa). When PSA is moderately increased (e.g., 4–10 ng/ml), it is difficult to differentiate benign prostatic hyperplasia (BPH) from cancer. The diagnostic test (i.e., prostate biopsy) is invasive, adding pain and economic burden to the patient. Urine samples are more convenient, non-invasive and readily available than blood. We sought to determine whether ferritin might be the potential urinary biomarker in prostate cancer diagnosis. Using two-dimensional electrophoresis (2DE) followed by mass spectrometry (MS), differentially expressed urinary proteins among patients with PCa, BPH and normal controls were obtained. The ferritin heavy chain (FTH) gene, ferritin light chain (FTL) gene and protein expression of BPH-1 cells and PC-3 cells were analyzed by real-time quantitative PCR and Western blotting, respectively. Stable FTH or FTL silenced cell lines were generated by small hairpin(sh) RNA lentiviral transfection. The function of the cell lines was evaluated by the colony formation assay, transwell assay, and flow cytometry. Compared with BPH and normal controls, 15 overexpressed proteins, including FTH and FTL, were identified in the urine of the PCa group. FTH and FTL were also highly expressed in PC-3 cell lines compared with BPH-1 cells. FTH-silenced cells showed reduced cell proliferation, migration and increased cell apoptosis. FTL-silenced cells showed increased proliferation and migration abilities. There are differences in urinary proteins among patients with PCa, BPH and normal controls. FTH and FTL play different roles in PCa cells and are potential biomarkers for PCa.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Siegel RL, Miller KD, Jemal A (2018) Cancer statistics, 2018. CA Cancer J Clin 68(1):7–30. https://doi.org/10.3322/caac.21442
Thompson IM (2004) Prevalence of prostate cancer among men with a prostate-specific antigen level <= 4.0 ng per milliliter (vol 350, pg 2239, 2004). N Engl J Med 351(14):1470–1470
Andriole GL, Levin DL, Crawford ED, Gelmann EP, Pinsky PF, Chia D, Kramer BS, Reding D, Church TR, Grubb RL, Izmirlian G, Ragard LR, Clapp JD, Prorok PC, Gohagan JK, Team PP (2005) Prostate Cancer screening in the prostate, lung, colorectal and ovarian (PLCO) Cancer screening trial: findings from the initial screening round of a randomized trial. J Natl Cancer Inst 97(6):433–438. https://doi.org/10.1093/jnci/dji065
Pieper R, Gatlin CL, McGrath AM, Makusky AJ, Mondal M, Seonarain M, Field E, Schatz CR, Estock MA, Ahmed N, Anderson NG, Steiner S (2004) Characterization of the human urinary proteome: a method for high-resolution display of urinary proteins on two-dimensional electrophoresis gels with a yield of nearly 1400 distinct protein spots. Proteomics 4(4):1159–1174
Adachi J, Kumar C, Zhang YL, Olsen JV, Mann M (2006) The human urinary proteome contains more than 1500 proteins, including a large proportion of membrane proteins. Genome Biol 7(9)
Crichton RR, Declercq JP (2010) X-ray structures of ferritins and related proteins. Biochim Biophys Acta 1800(8):706–718. https://doi.org/10.1016/j.bbagen.2010.03.019
Knovich MA, Storey JA, Coffman LG, Torti SV, Torti FM (2009) Ferritin for the clinician. Blood Rev 23(3):95–104. https://doi.org/10.1016/j.blre.2008.08.001
VanWagner LB, Green RM (2014) Elevated serum ferritin. JAMA 312(7):743–744. https://doi.org/10.1001/jama.2014.302
Alkhateeb AA, Connor JR (2013) The significance of ferritin in cancer: anti-oxidation, inflammation and tumorigenesis. Biochim Biophys Acta 1836(2):245–254. https://doi.org/10.1016/j.bbcan.2013.07.002
Cujic D, Golubovic S, Bojic-Trbojevic Z, Ilic N, Baricevic I, Nedic O (2010) Differential diagnosis of liver diseases using serum biomarkers. J BUON 15(1):141–146
Su Q, Lei T, Zhang M (2017) Association of ferritin with prostate cancer. J BUON 22(3):766–770
Meng Q, Lei T, Zhang M, Zhao J, Zhao XH, Zhang M (2013) Identification of proteins differentially expressed in adriamycin-resistant (pumc-91/ADM) and parental (pumc-91) human bladder cancer cell lines by proteome analysis. J Cancer Res Clin Oncol 139(3):509–519. https://doi.org/10.1007/s00432-012-1350-8
Ito K (2014) Prostate cancer in Asian men. Nat Rev Urol 11(4):197–212. https://doi.org/10.1038/nrurol.2014.42
Tabayoyong W, Abouassaly R (2015) Prostate Cancer screening and the associated controversy. Surg Clin N Am 95(5):1023. https://doi.org/10.1016/j.suc.2015.05.001
Wang XJ, An P, Zeng JL, Liu XY, Wang B, Fang XX, Wang FD, Ren GP, Min JX (2017) Serum ferritin in combination with prostate-specific antigen improves predictive accuracy for prostate cancer. Oncotarget 8(11):17862–17872
Arosio P, Levi S (2010) Cytosolic and mitochondrial ferritins in the regulation of cellular iron homeostasis and oxidative damage. Biochim Biophys Acta Gen Subj 1800(8):783–792. https://doi.org/10.1016/j.bbagen.2010.02.005
Boyd D, Vecoli C, Belcher DM, Jain SK, Drysdale JW (1985) Structural and functional relationships of human ferritin H and L chains deduced from cDNA clones. J Biol Chem 260(21):11755–11761
Arosio P, Adelman TG, Drysdale JW (1978) On ferritin heterogeneity. Further evidence for heteropolymers. J Biol Chem 253(12):4451–4458
Alkhateeb AA, Connor JR (2010) Nuclear ferritin: a new role for ferritin in cell biology. Biochim Biophys Acta Gen Subj 1800(8):793–797. https://doi.org/10.1016/j.bbagen.2010.03.017
Arosio P, Ingrassia R, Cavadini P (2009) Ferritins: a family of molecules for iron storage, antioxidation and more. Biochim Biophys Acta Gen Subj 1790(7):589–599. https://doi.org/10.1016/j.bbagen.2008.09.004
Kew MC, Torrance JD, Derman D, Simon M, Macnab GM, Charlton RW, Bothwell TH (1978) Serum and tumour ferritins in primary liver cancer. Gut 19(4):294–299
Eshhar Z, Order SE, Katz DH (1974) Ferritin, a Hodgkin's disease associated antigen. Proc Natl Acad Sci U S A 71(10):3956–3960
Alkhateeb AA, Han B, Connor JR (2013) Ferritin stimulates breast cancer cells through an iron-independent mechanism and is localized within tumor-associated macrophages. Breast Cancer Res Treat 137(3):733–744
Marcus DM, Zinberg N (1974) Isolation of ferritin from human mammary and pancreatic carcinomas by means of antibody immunoadsorbents. Arch Biochem Biophys 162(2):493–501
Arosio P, Yokota M, Drysdale JW (1976) Structural and immunological relationships of isoferritins in normal and malignant cells. Cancer Res 36(5):1735–1739
Torti FM, Torti SV (2002) Regulation of ferritin genes and protein. Blood 99(10):3505–3516
Theil EC (2011) Ferritin protein Nanocages use ion channels, catalytic sites, and nucleation channels to manage Iron/oxygen chemistry: a review for: current opinion in chemical biology/bioinorganic chemistry: Iron biochemistry. Curr Opin Chem Biol 15(2):304–311
Yang DC, Jiang X, Elliott RL, Head JF (2002) Antisense ferritin oligonucleotides inhibit growth and induce apoptosis in human breast carcinoma cells. Anticancer Res 22(3):1513–1524
Lobello N, Biamonte F, Pisanu ME, Faniello MC, Jakopin Ž, Chiarella E, Giovannone ED, Mancini R, Ciliberto G, Cuda G (2016) Ferritin heavy chain is a negative regulator of ovarian cancer stem cell expansion and epithelial to mesenchymal transition. Oncotarget 7(38):62019–62033
Min PJRC (2015) Role of ferritin in Cancer biology. J Cancer Sci Ther 07(5)
Li RS, Luo C, Mines M, Zhang JW, Fan GH (2006) Chemokine CXCL12 induces binding of ferritin heavy chain to the chemokine receptor CXCR4, alters CXCR4 signaling, and induces phosphorylation and nuclear translocation of ferritin heavy chain. J Biol Chem 281(49):37616–37627. https://doi.org/10.1074/jbc.M607266200
Coffman LG, Parsonage D, D'Agostino R, Torti FM, Torti SV (2009) Regulatory effects of ferritin on angiogenesis. Proc Natl Acad Sci U S A 106(2):570–575. https://doi.org/10.1073/pnas.0812010106
Cozzi A, Levi S, Corsi B, Santambrogio P, Campanella A, Gerardi G, Arosio P (2003) Role of iron and ferritin in TNFalpha-induced apoptosis in HeLa cells. FEBS Lett 537(1–3):187–192
Aung W, Hasegawa S, Furukawa T, Saga T (2007) Potential role of ferritin heavy chain in oxidative stress and apoptosis in human mesothelial and mesothelioma cells: implications for asbestos-induced oncogenesis. Carcinogenesis 28(9):2047–2052. https://doi.org/10.1093/carcin/bgm090
Coffman LG, Parsonage D, D'Agostino R Jr, Torti FM, Torti SV (2009) Regulatory effects of ferritin on angiogenesis. Proc Natl Acad Sci U S A 106(2):570–575. https://doi.org/10.1073/pnas.0812010106
Tesfay L, Huhn AJ, Hatcher H, Torti FM, Torti SV (2012) Ferritin blocks inhibitory effects of two-chain high molecular weight kininogen (HKa) on adhesion and survival signaling in endothelial cells. PLoS One 7(7):e40030. https://doi.org/10.1371/journal.pone.0040030
Surguladze N, Thompson KM, Beard JL, Connor JR, Fried MG (2004) Interactions and reactions of ferritin with DNA. J Biol Chem 279(15):14694–14702. https://doi.org/10.1074/jbc.M313348200
Thompson KJ, Fried MG, Ye Z, Boyer P, Connor JR (2002) Regulation, mechanisms and proposed function of ferritin translocation to cell nuclei. J Cell Sci 115(Pt 10):2165–2177
Ghosh S, Hevi S, Chuck SL (2004) Regulated secretion of glycosylated human ferritin from hepatocytes. Blood 103(6):2369–2376. https://doi.org/10.1182/blood-2003-09-3050
Ferring-Apel D, Hentze MW, Galy B (2009) Cell-autonomous and systemic context-dependent functions of iron regulatory protein 2 in mammalian iron metabolism. Blood 113(3):679–687. https://doi.org/10.1182/blood-2008-05-155093
Conflicts of interest
Huijun Zhao, Xuhong Zhao, Ting Lei, Man Zhang declare that they have no conflicts of interest.
This work was supported by the Beijing Municipal Administration of Hospitals’ Ascent Plan (DFL20150701) and the Beijing Natural Science Foundation (7172106).
All procedures performed in studies involving human participants were conducted in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. This article does not contain any studies with animals performed by any of the authors.
Informed consent was obtained from all individual participants included in the study.
About this article
Cite this article
Zhao, H., Zhao, X., Lei, T. et al. Screening, identification of prostate cancer urinary biomarkers and verification of important spots. Invest New Drugs 37, 935–947 (2019). https://doi.org/10.1007/s10637-018-0709-3
- Urinary proteomics
- Prostate cancer (PCa)
- Benign prostatic hyperplasia (BPH)
- RNA interference