Specific protein and miRNA patterns characterise tumour-associated fibroblasts in bladder cancer.
OBJECTIVE
Tumour development and progression are strongly affected by interaction of tumour cells and tumour stroma. Different tumour models demonstrate a supportive effect of tumour-associated fibroblasts (TAF) on the tumour genesis. Aims of the present study are the isolation of TAF from primary urinary bladder tumour specimens and the proteomic and epigenetic characterisation.


METHODS
TAF were isolated from cultured urinary bladder tumour specimens. Therefore, primary tumour material was treated with EDTA followed by two separated detachment steps. Non-tumour fibroblasts were isolated from foreskin and normal bladder tissues. Proteins and total RNA were isolated from cultured fibroblasts. Protein pattern analyses were carried out by SELDI-TOF-MS. The miRNA expression profile was analysed by miRNA microarray.


RESULTS
By optimising cell culture routines, we achieved to isolate and subsequently cultivate TAF from primary tumour material of the urinary bladder. SELDI-TOF-MS measurements reveal distinct differences in the proteomic patterns of TAF and non-tumour fibroblasts. Microarray analyses indicate specific expression of several miRNAs in TAF and non-tumour fibroblasts.


CONCLUSIONS
In summary, we determined proteomic and epigenetic differences between non-tumour fibroblasts and TAF of urinary bladder carcinoma and identified specific protein expression patterns as well as miRNA profiles of TAF in comparison with non-tumour fibroblasts. These findings provide more insights into the complex tumour network and a good starting point for the identification of markers for the prediction of tumour development and progression based on specific TAF expression patterns.
Enkelmann
Astrid
A
Laboratory of Molecular Biology, Department of Urology, Jena University Hospital, 07740, Jena, Germany. astrid.enkelmann@med.uni-jena.de
Heinzelmann
Joana
J
von Eggeling
Ferdinand
F
Walter
Martina
M
Berndt
Alexander
A
Wunderlich
Heiko
H
Junker
Kerstin
K
D045744:Cell Line, Tumor; D005347:Fibroblasts / Q000737:chemistry*; D006801:Humans; D035683:MicroRNAs / Q000032:analysis*; D009363:Neoplasm Proteins / Q000032:analysis*; D020411:Oligonucleotide Array Sequence Analysis; D040081:Protein Array Analysis; D040901:Proteomics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D018800:Thy-1 Antigens / Q000032:analysis; D001749:Urinary Bladder Neoplasms / Q000473:pathology*
D035683:MicroRNAs; D009363:Neoplasm Proteins; D018800:Thy-1 Antigens