title: Profilin 1 is a potential biomarker for bladder cancer aggressiveness.
abstract: Of the most important clinical needs for bladder cancer (BC) management is the identification of biomarkers for disease aggressiveness. Urine is a "gold mine" for biomarker discovery, nevertheless, with multiple proteins being in low amounts, urine proteomics becomes challenging. In the present study we applied a fractionation strategy of urinary proteins based on the use of immobilized metal affinity chromatography for the discovery of biomarkers for aggressive BC. Urine samples from patients with non invasive (two pools) and invasive (two pools) BC were subjected to immobilized metal affinity chromatography fractionation and eluted proteins analyzed by 1D-SDS-PAGE, band excision and liquid chromatography tandem MS. Among the identified proteins, multiple corresponded to proteins with affinity for metals and/or reported to be phosphorylated and included proteins with demonstrated association with BC such as MMP9, fibrinogen forms, and clusterin. In agreement to the immobilized metal affinity chromatography results, aminopeptidase N, profilin 1, and myeloblastin were further found to be differentially expressed in urine from patients with invasive compared with non invasive BC and benign controls, by Western blot or Elisa analysis, nevertheless exhibiting high interindividual variability. By tissue microarray analysis, profilin 1 was found to have a marked decrease of expression in the epithelial cells of the invasive (T2+) versus high risk non invasive (T1G3) tumors with occasional expression in stroma; importantly, this pattern strongly correlated with poor prognosis and increased mortality. The functional relevance of profilin 1 was investigated in the T24 BC cells where blockage of the protein by the use of antibodies resulted in decreased cell motility with concomitant decrease in actin polymerization. Collectively, our study involves the application of a fractionation method of urinary proteins and as one main result of this analysis reveals the association of profilin 1 with BC paving the way for its further investigation in BC stratification.
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens, Greece.
authors.forename: Jerome
authors.initials: J
authors.lastname: Zoidakis
authors.forename: Manousos
authors.initials: M
authors.lastname: Makridakis
authors.forename: Panagiotis G
authors.initials: PG
authors.lastname: Zerefos
authors.forename: Vasiliki
authors.initials: V
authors.lastname: Bitsika
authors.forename: Sergio
authors.initials: S
authors.lastname: Esteban
authors.forename: Maria
authors.initials: M
authors.lastname: Frantzi
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Stravodimos
authors.forename: Nikolaos P
authors.initials: NP
authors.lastname: Anagnou
authors.forename: Maria G
authors.initials: MG
authors.lastname: Roubelakis
authors.forename: Marta
authors.initials: M
authors.lastname: Sanchez-Carbayo
authors.forename: Antonia
authors.initials: A
authors.lastname: Vlahou
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000378:metabolism / Q000652:urine*; D018826:CD13 Antigens / Q000652:urine; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D004847:Epithelial Cells / Q000378:metabolism; D006801:Humans; D008875:Middle Aged; D053146:Myeloblastin / Q000652:urine; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000378:metabolism / Q000652:urine; D051304:Profilins / Q000378:metabolism / Q000652:urine*; D017154:Stromal Cells / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D001743:Urinary Bladder / Q000378:metabolism / Q000473:pathology; D001749:Urinary Bladder Neoplasms / Q000378:metabolism / Q000473:pathology / Q000652:urine*
chemical_list: D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; C495738:PFN1 protein, human; D051304:Profilins; D018826:CD13 Antigens; D053146:Myeloblastin