Differences in urinary proteins related to surgical margin status after radical prostatectomy

Heger,Zbynek; Michalek,Petr; Guran,Roman; Cernei,Natalia; Duskova,Katerina; Vesely,Stepan; Anyz,Jiri; Stepankova,Olga; Zitka,Ondrej; Adam,Vojtech; Kizek,Rene

doi:10.3892/or.2015.4322

Differences in urinary proteins related to surgical margin status after radical prostatectomy

Authors:
- Zbynek Heger
- Petr Michalek
- Roman Guran
- Natalia Cernei
- Katerina Duskova
- Stepan Vesely
- Jiri Anyz
- Olga Stepankova
- Ondrej Zitka
- Vojtech Adam
- Rene Kizek
View Affiliations

Affiliations: Central European Institute of Technology, Brno University of Technology, CZ-616 00 Brno, Czech Republic, Department of Urology, 2nd Faculty of Medicine, Charles University, and University Hospital Motol, CZ-150 06 Prague 5, Czech Republic, Department of Cybernetics, Faculty of Electrical Engineering, Czech Technical University, Prague, CZ 121 35 Prague 2, Czech Republic
Published online on: October 2, 2015 https://doi.org/10.3892/or.2015.4322
Pages: 3247-3255

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Presented exploratory pilot study was aimed at evaluation of proteins present in urinary specimens collected from prostate cancer suffering subjects after radical prostatectomy, divided into two experimental cohorts: positive (n=15) and negative (n=15) surgical margins (PSM/NSM). The presence of PSM suggests inadequate cancer clearance and the possible need for additional treatment. Proper identification of these risk-patients is therefore of a paramount importance. Total protein profiles were firstly identified by using SDS-PAGE and compared by using partial least square discrimination analysis (PLS-DA), which revealed differences in molecular weights of 80-99 and 150-235 kDa between the experimental groups. For further identification of proteins, comparative proteomic technologies were employed. Two-dimensional gel electrophoresis with subsequent identification of protein spots by using MALDI-TOF mass fingerprinting revealed differential expression of proteins between NSM/PSM cohorts. Moreover, in PSM group, three uniquely identified proteins (cyclin-dependent kinase 6, galectin-3-binding protein and L-lactate dehydrogenase C chain) were found, which show tight connection with prostate cancer and presence of all of them was previously linked to certain aspects of prostate cancer. These proteins may be associated with the molecular mechanisms of prostate cancer development; hence, their identification may be helpful for the assessment of disease progression risk after radical prostatectomy, but also for possible early diagnosis.

View Figures

View References

1	Arnold M, Karim-Kos HE, Coebergh JW, Byrnes G, Antilla A, Ferlay J, Renehan AG, Forman D and Soerjomataram I: recent trends in incidence of five common cancers in 26 European countries since 1988: Analysis of the European Cancer Observatory. Eur J Cancer. 51:1164–1187. 2015. View Article : Google Scholar
2	Luengo-Fernandez R, Leal J, Gray A and Sullivan R: Economic burden of cancer across the European Union: A population-based cost analysis. Lancet Oncol. 14:1165–1174. 2013. View Article : Google Scholar : PubMed/NCBI
3	Sakr WA, Haas GP, Cassin BF, Pontes JE and Crissman JD: The frequency of carcinoma and intraepithelial neoplasia of the prostate in young male patients. J Urol. 150:379–385. 1993.PubMed/NCBI
4	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2012. CA Cancer J Clin. 62:10–29. 2012. View Article : Google Scholar : PubMed/NCBI
5	Haas GP, Delongchamps N, Brawley OW, Wang CY and de la Roza G: The worldwide epidemiology of prostate cancer: Perspectives from autopsy studies. Can J Urol. 15:3866–3871. 2008.PubMed/NCBI
6	Walsh PC, Partin AW and Epstein JI: Cancer control and quality of life following anatomical radical retropubic prostatectomy: Results at 10 years. J Urol. 152:1831–1836. 1994.PubMed/NCBI
7	Watson RB, Civantos F and Soloway MS: Positive surgical margins with radical prostatectomy: Detailed pathological analysis and prognosis. Urology. 48:80–90. 1996. View Article : Google Scholar : PubMed/NCBI
8	Cheng L, Darson MF, Bergstralh EJ, Slezak J, Myers RP and Bostwick DG: Correlation of margin status and extraprostatic extension with progression of prostate carcinoma. Cancer. 86:1775–1782. 1999. View Article : Google Scholar : PubMed/NCBI
9	Vesely S, Jarolim L, Duskova K, Schmidt M, Dusek P and Babjuk M: The use of early postoperative prostate-specific antigen to stratify risk in patients with positive surgical margins after radical prostatectomy. BMC Urol. 14:792014. View Article : Google Scholar : PubMed/NCBI
10	Aebersold R, Anderson L, Caprioli R, Druker B, Hartwell L and Smith R: Perspective: A program to improve protein biomarker discovery for cancer. J Proteome Res. 4:1104–1109. 2005. View Article : Google Scholar : PubMed/NCBI
11	Bergman N and Bergquist J: Recent developments in proteomic methods and disease biomarkers. Analyst (Lond). 139:3836–3851. 2014. View Article : Google Scholar
12	Heger Z, Kominkova M, Cernei N, Krejcova L, Kopel P, Zitka O, Adam V and Kizek R: Fluorescence resonance energy transfer between green fluorescent protein and doxorubicin enabled by DNA nanotechnology. Electrophoresis. 35:3290–3301. 2014. View Article : Google Scholar : PubMed/NCBI
13	Vyslouzilova L, Krizkova S, Anyz J, Hynek D, Hrabeta J, Kruseova J, Eckschlager T, Adam V, Stepankova O and Kizek R: Use of brightness wavelet transformation for automated analysis of serum metallothioneins- and zinc-containing proteins by western blots to subclassify childhood solid tumours. Electrophoresis. 34:1637–1648. 2013. View Article : Google Scholar : PubMed/NCBI
14	Shevchenko A, Tomas H, Havlis J, Olsen JV and Mann M: In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nat Protoc. 1:2856–2860. 2006. View Article : Google Scholar
15	Oh J, Pyo JH, Jo EH, Hwang SI, Kang SC, Jung JH, Park EK, Kim SY, Choi JY and Lim J: Establishment of a near-standard two-dimensional human urine proteomic map. Proteomics. 4:3485–3497. 2004. View Article : Google Scholar : PubMed/NCBI
16	Rehman I, Azzouzi AR, Catto JWF, Allen S, Cross SS, Feeley K, Meuth M and Hamdy FC: Proteomic analysis of voided urine after prostatic massage from patients with prostate cancer: A pilot study. Urology. 64:1238–1243. 2004. View Article : Google Scholar : PubMed/NCBI
17	Ye B, Skates S, Mok SC, Horick NK, Rosenberg HF, Vitonis A, Edwards D, Sluss P, Han WK, Berkowitz RS, et al: Proteomic-based discovery and characterization of glycosylated eosinophil-derived neurotoxin and COOH-terminal osteopontin fragments for ovarian cancer in urine. Clin Cancer Res. 12:432–441. 2006. View Article : Google Scholar : PubMed/NCBI
18	Botezatu I, Serdyuk O, Potapova G, Shelepov V, Alechina R, Molyaka Y, Ananév V, Bazin I, Garin A, Narimanov M, et al: Genetic analysis of DNA excreted in urine: A new approach for detecting specific genomic DNA sequences from cells dying in an organism. Clin Chem. 46:1078–1084. 2000.PubMed/NCBI
19	Oh J, Wilson L, Kirk M, Deshane J and Kim H: Proteomics of normal human and prostate cancer urines. FASEB J. 17:759.7572003.
20	Kshirsagar B and Wiggins RC: A map of urine proteins based on one-dimensional SDS-polyacrylamide gel electrophoresis and western blotting using one microliter of unconcentrated urine. Clin Chim Acta. 158:13–22. 1986. View Article : Google Scholar : PubMed/NCBI
21	Zaragoza C, Barrera R, Centeno F, Tapia JA and Mañe MC: Canine pyometra: A study of the urinary proteins by SDS-PAGE and western blot. Theriogenology. 61:1259–1272. 2004. View Article : Google Scholar : PubMed/NCBI
22	Le Bricon T, Erlich D, Bengoufa D, Dussaucy M, Garnier JP and Bousquet B: Sodium dodecyl sulfate-agarose gel electrophoresis of urinary proteins: Application to multiple myeloma. Clin Chem. 44:1191–1197. 1998.PubMed/NCBI
23	Marshall T and Williams KM: High resolution two-dimensional electrophoresis of human urinary proteins. Anal Chim Acta. 372:147–160. 1998. View Article : Google Scholar
24	Thongboonkerd V, McLeish KR, Arthur JM and Klein JB: Proteomic analysis of normal human urinary proteins isolated by acetone precipitation or ultracentrifugation. Kidney Int. 62:1461–1469. 2002. View Article : Google Scholar : PubMed/NCBI
25	Adachi J, Kumar C, Zhang Y, Olsen JV and Mann M: The human urinary proteome contains more than 1500 proteins, including a large proportion of membrane proteins. Genome Biol. 7:R802006. View Article : Google Scholar : PubMed/NCBI
26	Day ML, Zhao X, Vallorosi CJ, Putzi M, Powell CT, Lin C and Day KC: E-cadherin mediates aggregation-dependent survival of prostate and mammary epithelial cells through the retinoblastoma cell cycle control pathway. J Biol Chem. 274:9656–9664. 1999. View Article : Google Scholar : PubMed/NCBI
27	Cheng L, Nagabhushan M, Pretlow TP, Amini SB and Pretlow TG: Expression of E-cadherin in primary and metastatic prostate cancer. Am J Pathol. 148:1375–1380. 1996.PubMed/NCBI
28	Umbas R, Isaacs WB, Bringuier PP, Schaafsma HE, Karthaus HF, Oosterhof GO, Debruyne FM and Schalken JA: Decreased E-cadherin expression is associated with poor prognosis in patients with prostate cancer. Cancer Res. 54:3929–3933. 1994.PubMed/NCBI
29	Giroldi LA and Schalken JA: Decreased expression of the intercellular adhesion molecule E-cadherin in prostate cancer: Biological significance and clinical implications. Cancer Metastasis Rev. 12:29–37. 1993. View Article : Google Scholar : PubMed/NCBI
30	Shimamura T, Sakamoto M, Ino Y, Sato Y, Shimada K, Kosuge T, Sekihara H and Hirohashi S: Dysadherin overexpression in pancreatic ductal adenocarcinoma reflects tumor aggressiveness: Relationship to e-cadherin expression. J Clin Oncol. 21:659–667. 2003. View Article : Google Scholar : PubMed/NCBI
31	Duffy MJ: Tumor markers in clinical practice: A review focusing on common solid cancers. Med Princ Pract. 22:4–11. 2013. View Article : Google Scholar
32	Gutman EB, Sproul EE and Gutman AB: Significance of increased phosphatase activity of bone at the site of osteoplastic metastases secondary to carcinoma of the prostate gland. Am J Cancer. 28:485–495. 1936. View Article : Google Scholar
33	Wang MC, Papsidero LD, Kuriyama M, Valenzuela LA, Murphy GP and Chu TM: Prostate antigen: A new potential marker for prostatic cancer. Prostate. 2:89–96. 1981. View Article : Google Scholar : PubMed/NCBI
34	Morikawa W, Yamamoto K, Ishikawa S, Takemoto S, Ono M, Fukushi J, Naito S, Nozaki C, Iwanaga S and Kuwano M: Angiostatin generation by cathepsin D secreted by human prostate carcinoma cells. J Biol Chem. 275:38912–38920. 2000. View Article : Google Scholar : PubMed/NCBI
35	Lim JTE, Mansukhani M and Weinstein IB: Cyclin-dependent kinase 6 associates with the androgen receptor and enhances its transcriptional activity in prostate cancer cells. Proc Natl Acad Sci USA. 102:5156–5161. 2005. View Article : Google Scholar : PubMed/NCBI
36	Choi YJ and Anders L: Signaling through cyclin D-dependent kinases. Oncogene. 33:1890–1903. 2014. View Article : Google Scholar
37	Hanahan D and Weinberg RA: Hallmarks of cancer: The next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
38	Finn RS, Dering J, Conklin D, Kalous O, Cohen DJ, Desai AJ, Ginther C, Atefi M, Chen I, Fowst C, et al: PD 0332991, a selective cyclin D kinase 4/6 inhibitor, preferentially inhibits proliferation of luminal estrogen receptor-positive human breast cancer cell lines in vitro. Breast Cancer Res. 11:R772009. View Article : Google Scholar : PubMed/NCBI
39	Tan MHE, Li J, Xu HE, Melcher K and Yong EL: Androgen receptor: Structure, role in prostate cancer and drug discovery. Acta Pharmacol Sin. 36:3–23. 2015. View Article : Google Scholar :
40	Sardana G, Marshall J and Diamandis EP: Discovery of candidate tumor markers for prostate cancer via proteomic analysis of cell culture-conditioned medium. Clin Chem. 53:429–437. 2007. View Article : Google Scholar : PubMed/NCBI
41	Inohara H, Akahani S, Koths K and Raz A: Interactions between galectin-3 and Mac-2-binding protein mediate cell-cell adhesion. Cancer Res. 56:4530–4534. 1996.PubMed/NCBI
42	Hu J, He J, Kuang Y, Wang Z, Sun Z, Zhu H and Liu X: Expression and significance of 90k/Mac-2BP in prostate cancer. Exp Ther Med. 5:181–184. 2013.
43	Srirajaskanthan R, Caplin ME, Waugh MG, Watkins J, Meyer T, Hsuan JJ and Beaumont NJ: Identification of Mac-2-binding protein as a putative marker of neuroendocrine tumors from the analysis of cell line secretomes. Mol Cell Proteomics. 9:656–666. 2010. View Article : Google Scholar :
44	Scanlan MJ, Simpson AJG and Old LJ: The cancer/testis genes: Review, standardization, and commentary. Cancer Immun. 4:1–15. 2004.PubMed/NCBI
45	Gure AO, Chua R, Williamson B, Gonen M, Ferrera CA, Gnjatic S, Ritter G, Simpson AJ, Chen YT, Old LJ, et al: Cancer-testis genes are coordinately expressed and are markers of poor outcome in non-small cell lung cancer. Clin Cancer Res. 11:8055–8062. 2005. View Article : Google Scholar : PubMed/NCBI
46	Andrade VCC, Vettore AL, Almeida MSS, et al: Prognostic impact of cancer testis antigens expression in advanced stage multiple myeloma patients. Blood. 110:257B. 2007.
47	Marcovich R, Wojno KJ, Wei JT, Rubin MA, Montie JE and Sanda MG: Bladder neck-sparing modification of radical prostatectomy adversely affects surgical margins in pathologic T3a prostate cancer. Urology. 55:904–908. 2000. View Article : Google Scholar : PubMed/NCBI