Change search
ReferencesLink to record
Permanent link

Direct link
Proteomic Analysis of Urinary Bladder Cancer: Aiming for Novel Biomarkers
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Surgical Sciences, Urology. (Experimentell Urologi)
2013 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Urinary bladder cancer is a heterogeneous disease appearing in different forms, e.g. non-muscle invasive and muscle invasive. For all variants, the expression of proteins is interesting to analyze for diagnostic, predictive, prognostic and drug targeting purposes, since it reflects the altered gene expression causing the cancer. Since urothelial cells of the bladder are in direct contact with urine it is likely that this body fluid contains cancer-related proteins. In Paper I, unbiased analysis of proteins in urine from urinary bladder cancer patients and controls, using label-free quantification by mass spectrometry, was applied and four interesting proteins APOE, FGB, LRG and SERPINA1 were selected and further analyzed with western and dot blot. In Paper II, two more proteins, POLR1E and TOP2A, were validated as relevant proteins in bladder cancer urine. In Paper III and IV, the proteins GAL1 and STMN1 were investigated for their prognostic and therapeutic target potential in bladder cancer. In Paper II, III and IV, the expression of seven of the proteins were analyzed on tissue microarrays representing tumour tissue from 360 patients with different tumour stages. For the proteins identified by the urine screening approach, their protein expressions were confirmed in bladder cancer tissue. The expression level in tissue of five of the proteins, APOE, FGB, POLR1E (Paper II), GAL1 (Paper III) and STMN1 (Paper IV), increased with tumour stage, showing diagnostic relevance and three of the proteins, SERPINA1 (Paper II), STMN1 (Paper IV) and GAL1 (Paper III) had prognostic potential in urinary bladder cancer. In addition, GAL1 and STMN1 were demonstrated to be highly expressed in metastatic disease and inhibition of STMN1 reduced cell growth (Paper III and IV), indicating that these proteins are promising drug targets in urinary bladder cancer. In conclusion, the approach of this thesis has generated several candidate protein biomarkers in urine and tissue, validated with independent methods, which have the potential to improve the care for bladder cancer patients.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2013. , 61 p.
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 879
Keyword [en]
urine, APOE, FGB, GAL1, LRG1, POLR1E, SERPINA1, STMN1, TOP2A, biomarker, diagnostic biomarker, prognostic biomarker, predictive biomarker, mass spectrometry, western blot, dot blot, immunohistochemistry, IHC, tissue microarray, TMA, urothelium, antibody, antibody-based
Keyword [sv]
urinblåsecancer, biomarkör, diagnos, prognos, urin, proteomik, masspektrometri, immunohistokemi, antikroppar
National Category
Medical and Health Sciences
Research subject
Molecular Biology; Molecular Medicine
URN: urn:nbn:se:uu:diva-197132ISBN: 978-91-554-8625-9OAI: diva2:611903
Public defence
2013-05-03, The Rudbeck Hall, Dag Hammarskjölds väg 20, Uppsala, 13:00 (English)
Available from: 2013-04-11 Created: 2013-03-18 Last updated: 2013-08-30Bibliographically approved
List of papers
1. Proteomic analysis of urinary biomarker candidates for nonmuscle invasive bladder cancer
Open this publication in new window or tab >>Proteomic analysis of urinary biomarker candidates for nonmuscle invasive bladder cancer
Show others...
2012 (English)In: Proteomics, ISSN 1615-9853, E-ISSN 1615-9861, Vol. 12, no 1, 135-144 p.Article in journal (Refereed) Published
Abstract [en]

Nonmuscle invasive tumors of the bladder often recur and thereby bladder cancer patients need regular re-examinations which are invasive, unpleasant, and expensive. A noninvasive and less expensive method, e.g. a urine dipstick test, for monitoring recurrence would thus be advantageous. In this study, the complementary techniques mass spectrometry (MS) and Western blotting (WB)/dot blot (DB) were used to screen the urine samples from bladder cancer patients. High resolving MS was used to analyze and quantify the urinary proteome and 29 proteins had a significantly higher abundance (p<0.05) in bladder cancer samples compared with control urine samples. The increased abundance found in urine from bladder cancer patients compared with controls was confirmed with Western blot for four selected proteins; fibrinogen β chain precursor, apolipoprotein E, α-1-antitrypsin, and leucine-rich α-2-glycoprotein 1. Dot blot analysis of an independent urine sample set pointed out fibrinogen β chain and α-1-antitrypsin as most interesting biomarkers having sensitivity and specificity values in the range of 66-85%. Exploring the Human Protein Atlas (HPA) also revealed that bladder cancer tumors are the likely source of these proteins. They have the potential of being useful in diagnosis, monitoring of recurrence and thus may improve the treatment of bladder tumors, especially nonmuscle invasive tumors.

National Category
Urology and Nephrology Cancer and Oncology
urn:nbn:se:uu:diva-163407 (URN)10.1002/pmic.201000810 (DOI)000298841000016 ()22065568 (PubMedID)
Available from: 2011-12-12 Created: 2011-12-12 Last updated: 2015-08-11Bibliographically approved
2. Tumour expression of bladder cancer-associated urinary proteins
Open this publication in new window or tab >>Tumour expression of bladder cancer-associated urinary proteins
Show others...
2013 (English)In: BJU International, ISSN 1464-4096, E-ISSN 1464-410X, Vol. 112, no 3, 407-415 p.Article in journal (Refereed) Published
Abstract [en]


  • The current basis for diagnosis and prognosis in urinary bladder cancer is based on the pathologists' assessment of a biopsy of the tumour. Urinary biomarkers are preferable as they can be non-invasively sampled. Urinary cytology is the only test with widespread use but is hampered by poor reproducibility and low sensitivity.
  • By studying the protein expression in bladder tumour tissue samples of proteins previously found in elevated levels in the urine of patients with bladder cancer, we have been able to show that these proteins originate from the tumour. The immunoreactivity of three of the investigated proteins increased with higher stage. Also a serine peptidase inhibitor was found to be predictive of progression from non-muscle-invasive to muscle-invasive tumours.


  • To analyse the expression of five bladder cancer-associated urinary proteins and investigate if expression is related to the malignant phenotype of the tumour.
  • To explore the possible prognostic value of these proteins.


  • Urine samples, 16 from patients with bladder cancer and 26 from controls, were used in Western Blotting experiments.
  • Tissue microarrays with bladder tissue from 344 patients diagnosed with bladder cancer between 1984 and 2005 was used in immunohistochemistry experiments.
  • The proteins apolipoprotein E (APOE), fibrinogen β chain precursor (FGB), leucine-rich α2-glycoprotein (LRG1), polymerase (RNA) I polypeptide E (POLR1E), α1-antitrypsin (SERPINA1) and topoisomerase 2A (TOP2A) were probed with antibodies validated by the Human Protein Atlas.


  • Increased expressions of APOE, FGB and POLR1E were correlated with increased tumour stage (P < 0.001).
  • Expression of SERPINA1 in Ta and T1 tumours was found to increase the risk of tumour progression (hazard ratio 2.57, 95% confidence interval 1.13-5.87; P = 0.025)


  • All proteins previously detected in urine from patients with bladder cancer were also expressed in bladder cancer tissue.
  • The expression of APOE, FGB and POLR1E increased with stage and they are potential diagnostic markers.
  • SERPINA1 was identified as a prognostic marker candidate.
National Category
Medical and Health Sciences
urn:nbn:se:uu:diva-197160 (URN)10.1111/j.1464-410X.2012.11653.x (DOI)000321429800024 ()23470167 (PubMedID)

De två sista författarna delar sistaförfattarskapet.

Available from: 2013-03-18 Created: 2013-03-18 Last updated: 2013-12-17Bibliographically approved
3. Galectin-1, a potential therapeutic target, in primary tumors and metastases of urinary bladder carcinomas
Open this publication in new window or tab >>Galectin-1, a potential therapeutic target, in primary tumors and metastases of urinary bladder carcinomas
Show others...
2013 (English)Manuscript (preprint) (Other academic)
Abstract [en]

Urinary bladder cancer would gain from new protein biomarkers due to the heterogeneity of disease. The beta-galactoside-binding protein (GAL1) is one such candidate and in present study its prognostic value and expression at protein level in metastatic bladder cancer-disease, have been evaluated. The protein expression of GAL1 was investigated by immunohistochemistry in two tumor cohorts, one with primary tumors of different stage and grade (n=344) and another with primary tumors matched with metastases (n=90). The expression in the actual cancer cells as well as in stroma and blood vessels were considered since the presence of GAL1 in different tissue compartments has shown cancer relevance. The cellular expression increased with increased tumor stage and grade (p<0.001). For the majority of the patients, cells from both primary tumor and metastasis showed a positive immunoreactivity for GAL1 (91% (n=64) for primary tumors with single metastasis (n=70) and 100% (n=20) for primary tumors with multiple metastasis (n=20). Further, strong immunoreactivity in T1 tumor cells correlated with lower risk of recurrence (p<0.05). Both tumors and metastasis exhibited strong stromal-GAL1 staining that could not be correlated with clinical parameters. The expression in vessels showed that T1 tumors surrounded by GAL1 negative blood vessels had a higher risk of progression (p<0.0001) into muscle invasive T2-4 stages. The results show that GAL1 is an important bladder cancer-protein from several aspects. Further, GAL1 is a promising therapeutic target in bladder cancer due to the general expression in advanced disease.

National Category
Medical and Health Sciences
urn:nbn:se:uu:diva-197192 (URN)
Available from: 2013-03-18 Created: 2013-03-18 Last updated: 2015-06-24
4. Stathmin-1 is a promising prognostic factor and potential therapeutic target in urinary bladder cancer
Open this publication in new window or tab >>Stathmin-1 is a promising prognostic factor and potential therapeutic target in urinary bladder cancer
Show others...
(English)Manuscript (preprint) (Other academic)
Abstract [en]

Aim: The oncoprotein 18/stathmin 1 (STMN1), involved in cell cycle progression and cell migration, has been reported to be expressed in several types of cancer, and is associated with clinical outcome in e.g. breast and liver cancer. The aims in this study were to investigate the clinical significance of STMN1 and to examine if STMN1 might be a possible therapeutic target in urinary bladder cancer.

Experimental design: Immunohistochemical analyses of STMN1 protein expression were performed in a wide-range tissue microarray (115 Ta-, 115 T1-, 112 T2-4-tumors) and in a metastatic primary tumor/matched metastasis-material (90 patients). In the T24 cell line, the effect of STMN1 on cell proliferation was evaluated by inhibiting the cellular expression of STMN using STMN1-siRNA.

Results: Patients with T1- or muscle-invasive disease exhibiting high expression of the STMN1 protein had a poorer overall survival (OS) and disease specific survival (DSS). In a multivariate analysis adjusting for stage, age and gender the results were for T2-T4 patients: OS (HR=1.77 95% CI 1.02-3.07; p=0.04) and DSS (HR=2.04 95% CI 1.13-3.68; p=0.02); for T1-4 patients: DSS (HR=1.83 95% CI 1.09-3.08; p=0.02). In the metastatic bladder cancer material, the majority of the patients with one metastasis (69%) and with several matched metastases (70%) were STMN1-positive in both the primary tumor and the matched metastases. Moreover, the ability of the urinary bladder cancer cell line to grow was significantly reduced after 72 hours (p<0.0001) when transfecting the cells with a siRNA targeting STMN1.

Conclusion: Our results suggest that STMN1 protein-expression has a potential both as a prognostic marker and a novel treatment target in urinary bladder cancer.

National Category
Medical and Health Sciences
Research subject
Medical Science
urn:nbn:se:uu:diva-197155 (URN)
Available from: 2013-03-18 Created: 2013-03-18 Last updated: 2015-06-24

Open Access in DiVA

fulltext(34585 kB)1238 downloads
File information
File name FULLTEXT01.pdfFile size 34585 kBChecksum SHA-512
Type fulltextMimetype application/pdf
Buy this publication >>

Search in DiVA

By author/editor
Lindén, Mårten
By organisation
Medical and Health Sciences

Search outside of DiVA

GoogleGoogle Scholar
Total: 1238 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Total: 1366 hits
ReferencesLink to record
Permanent link

Direct link