Abstract 547: Changes in proteomic and epigenetic expression patterns in tumour associated fibroblasts (TAF) by interaction with urinary bladder carcinoma cells

Abstract

Abstract Background: Tumour development and progression is strongly affected by interaction of tumour cells and tumour stroma. For different tumour models (e.g. breast cancer) a supportive effect of TAF on the tumour genesis was demonstrated. Purpose of the present work is the isolation and characterisation of TAF from primary urinary bladder tumour specimen. A further part of this study will deal with the influence of urinary bladder carcinoma cell lines on protein expression of TAF. Material and Methods: TAF were isolated from cultured urinary bladder tumour specimen. Therefore, primary tumour material was treated with EDTA followed by differential trypsinisation. Non-tumour fibroblasts were isolated from foreskin and normal urinary bladder tissue. Analyses of protein patterns were carried out on cultivated fibroblasts by SELDI-TOF-MS. TAF and foreskin fibroblasts were co-cultivated with urinary bladder cancer cell lines in separated cell culture compartments followed by SELDI-TOF-MS analyses. Furthermore total RNA was isolated from TAF and non-tumour fibroblasts to analyse the miRNA expression profile by miRNA Microarray. Results: By optimizing cell culture routines it was possible to isolate and subsequently cultivate TAF from primary tumour material of the urinary bladder. SELDI-TOF-MS measurements reveal differences in the proteomic patterns of TAF and non-tumour fibroblasts. Microarray analyses indicate different expression levels of several miRNAs in TAF and non-tumour fibroblasts. Co-cultivation of urinary bladder carcinoma cells and TAF or non-tumour fibroblasts induces modified protein patterns in the different cell types. Conclusion: TAF can be isolated and cultivated separately from primary tumour material. TAF are characterised by the expression of a specific protein pattern and miRNA profile in comparison to non-tumour fibroblasts. Co-cultivation with tumour cells revealed the induction of a modified expression profile in fibroblasts and vice versa. The present results will provide a more detailed understanding of the function of TAF in the tumour development of urinary bladder carcinoma. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 547.