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Characterization of human bladder cell membrane during cancer transformation.

Dobrzyńska I, Szachowicz-Petelska B, Darewicz B, Figaszewski ZA - J. Membr. Biol. (2015)

Bottom Line: The surface charge density of human bladder cell membranes was determined using electrophoresis.Our results show that cancer transformation is associated with increased phospholipid levels and a decreased level of integral proteins.In conclusion, this study demonstrates that cell membrane structure and function are modified in bladder cancer cells and that further work in this area is warranted.

View Article: PubMed Central - PubMed

Affiliation: Institute of Chemistry, University in Białystok, Al. Piłsudskiego 11/4, 15-443, Białystok, Poland, izadob@uwb.edu.pl.

ABSTRACT
Phenomena associated with changes in cell membranes are thought to play an important role in the cancer transformation. We hypothesized that the electrical charge of tumor cells can indirectly represent membrane-based changes that have occurred during cell transformation and may indicate tumor cell status. Here, we describe work showing that phospholipids, proteins content, and electric charge, are all altered in the cell membranes of pT2 stage/grade G3 bladder cancer. Qualitative and quantitative phospholipid composition and the presence of integral membrane proteins were identified using high-performance liquid chromatography. Protein composition was determined using selective hydrolysis of isolated bladder cell membrane proteins and peptide resolution. The surface charge density of human bladder cell membranes was determined using electrophoresis. Our results show that cancer transformation is associated with increased phospholipid levels and a decreased level of integral proteins. Moreover, the process of cancer transformation significantly enhanced changes in the surface charge density of the human bladder cell membrane. In conclusion, this study demonstrates that cell membrane structure and function are modified in bladder cancer cells and that further work in this area is warranted.

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Related in: MedlinePlus

A characteristic example of the separation of peptides from integral membrane proteins from bladder tissue (UV detected at 220 nm)
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Fig1: A characteristic example of the separation of peptides from integral membrane proteins from bladder tissue (UV detected at 220 nm)

Mentions: The membrane protein extracts were hydrolyzed using trypsin, at an enzyme:substrate ratio of 1:25 (Persaud et al. 2000). Following hydrolysis, the peptides were separated by HPLC on a LichroCART RP-18 column 100 A (5 μm, 250 × 4.0 mm) equilibrated with solvent A [0.1 % trifluoroacetic acid (TFA) in H2O] and eluted with a linear gradient of 20–100 % solvent B (0.1 % TFA in acetonitrile) using a flow rate of 1 mL/min (Szachowicz-Petelska et al. 2010). A typical separation of the peptide mixture containing integral bladder membrane proteins is provided in Fig. 1.Fig. 1


Characterization of human bladder cell membrane during cancer transformation.

Dobrzyńska I, Szachowicz-Petelska B, Darewicz B, Figaszewski ZA - J. Membr. Biol. (2015)

A characteristic example of the separation of peptides from integral membrane proteins from bladder tissue (UV detected at 220 nm)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4381039&req=5

Fig1: A characteristic example of the separation of peptides from integral membrane proteins from bladder tissue (UV detected at 220 nm)
Mentions: The membrane protein extracts were hydrolyzed using trypsin, at an enzyme:substrate ratio of 1:25 (Persaud et al. 2000). Following hydrolysis, the peptides were separated by HPLC on a LichroCART RP-18 column 100 A (5 μm, 250 × 4.0 mm) equilibrated with solvent A [0.1 % trifluoroacetic acid (TFA) in H2O] and eluted with a linear gradient of 20–100 % solvent B (0.1 % TFA in acetonitrile) using a flow rate of 1 mL/min (Szachowicz-Petelska et al. 2010). A typical separation of the peptide mixture containing integral bladder membrane proteins is provided in Fig. 1.Fig. 1

Bottom Line: The surface charge density of human bladder cell membranes was determined using electrophoresis.Our results show that cancer transformation is associated with increased phospholipid levels and a decreased level of integral proteins.In conclusion, this study demonstrates that cell membrane structure and function are modified in bladder cancer cells and that further work in this area is warranted.

View Article: PubMed Central - PubMed

Affiliation: Institute of Chemistry, University in Białystok, Al. Piłsudskiego 11/4, 15-443, Białystok, Poland, izadob@uwb.edu.pl.

ABSTRACT
Phenomena associated with changes in cell membranes are thought to play an important role in the cancer transformation. We hypothesized that the electrical charge of tumor cells can indirectly represent membrane-based changes that have occurred during cell transformation and may indicate tumor cell status. Here, we describe work showing that phospholipids, proteins content, and electric charge, are all altered in the cell membranes of pT2 stage/grade G3 bladder cancer. Qualitative and quantitative phospholipid composition and the presence of integral membrane proteins were identified using high-performance liquid chromatography. Protein composition was determined using selective hydrolysis of isolated bladder cell membrane proteins and peptide resolution. The surface charge density of human bladder cell membranes was determined using electrophoresis. Our results show that cancer transformation is associated with increased phospholipid levels and a decreased level of integral proteins. Moreover, the process of cancer transformation significantly enhanced changes in the surface charge density of the human bladder cell membrane. In conclusion, this study demonstrates that cell membrane structure and function are modified in bladder cancer cells and that further work in this area is warranted.

Show MeSH
Related in: MedlinePlus