Adipozyten induzieren bestimmte Genexpressionsmuster in Brustkrebszelllinien und verstärken die inflammatorische NfkB-Signalgebung in triple-negativen Brustkrebszellen

Obesity is a known risk factor for breast cancer. Since obesity rates are constantly rising worldwide, understanding the molecular details of the interaction between adipose tissue and breast tumors becomes an urgent task. To investigate potential molecular changes in breast cancer cells induced by co-existing adipocytes, we used a co-culture system of different breast cancer cell lines (MCF-7 and T47D: ER+/PR+/HER2- and MDA-MB-231: ER-/PR-/HER2-) and murine 3T3-L1 adipocytes. Here, we report that co-culture with adipocytes revealed distinct changes in global gene expression pattern in the different breast cancer cell lines. Our microarray data revealed that in both ER+ cell lines, top upregulated genes showed significant enrichment for hormone receptor target genes. In triple-negative MDA-MB-231 cells, co-culture with adipocytes led to the induction of pro-inflammatory genes, mainly involving genes of the Nf-κB signaling pathway. Moreover, co-cultured MDA-MB-231 cells showed increased secretion of the pro-inflammatory interleukins IL-6 and IL-8. Using a specific NF-κB inhibitor, these effects were significantly decreased. Finally, migratory capacities were significantly increased in triple-negative breast cancer cells upon co-culture with adipocytes, indicating an enhanced aggressive cell phenotype. Together, our studies illustrate that factors secreted by adipocytes have a significant impact on the molecular biology of breast cancer cells.:1. Einleitung ......................................................................................................... 4 1.1 Übergewicht und Adipositas – die Volkskrankheit des 21. Jahrhunderts ........ 4 1.2 Fettgewebe – Fett ist nicht gleich Fett ........................................................... 5 1.3 Fehlfunktionen des Fettgewebes bei Adipositas ............................................ 6 1.4 Der Zusammenhang zwischen Adipositas und Krebs ..................................... 7 1.4.1 Adipositas und die Tumormikroumgebung .................................................. 7 1.4.2 Adipositas-induzierte Inflammation und Krebs ............................................. 9 1.4.3 Die Insulin-IGF1-Achse................................................................................11 1.4.4 Östrogen-Synthese in adipösem Fettgewebe ............................................12 1.4.5 Tumorzellmetabolismus ..............................................................................12 1.5 Ziele der Arbeit ............................................................................................. 14 2 Publikation ...................................................................................................... 15 3 Zusammenfassung der Arbeit ......................................................................... 29 4 Literaturverzeichnis.......................................................................................... 34 5 Anlagen ........................................................................................................... 43 5.1 Supplemental Material .................................................................................. 43 5.1.1 Supplemental Figures ................................................................................45 5.1.2 Supplemental Tables ..................................................................................51 5.2 Erklärung über den wissenschaftlichen Beitrag des Promovenden zur Publikation ......................................................................................................... 66 5.3 Verzeichnis der verwendeten Abkürzungen und Symbole ............................ 67 5.4 Erklärung über die eigenständige Abfassung der Arbeit .............................. 69 5.5 Lebenslauf ................................................................................................... 70 5.6 Publikationsverzeichnis ................................................................................ 72 5.7 Vorträge und Posterpräsentationen mit publizierten Abstracts ..................... 73 5.8 Danksagung ................................................................................................. 7

Indirect p53-dependent transcriptional repression of Survivin, CDC25C, and PLK1 genes requires the cyclin-dependent kinase inhibitor p21/CDKN1A and CDE/CHR promoter sites binding the DREAM complex

The transcription factor p53 is central to cell cycle control by downregulation of cell cycle-promoting genes upon cell stress such as DNA damage. Survivin (BIRC5), CDC25C, and PLK1 encode important cell cycle regulators that are repressed following p53 activation. Here, we provide evidence that p53-dependent repression of these genes requires activation of p21 (CDKN1A, WAF1, CIP1). Chromatin immunoprecipitation (ChIP) data indicate that promoter binding of B-MYB switches to binding of E2F4 and p130 resulting in a replacement of the MMB (Myb-MuvB) by the DREAM complex. We demonstrate that this replacement depends on p21. Furthermore, transcriptional repression by p53 requires intact DREAM binding sites in the target promoters. The CDE and CHR cell cycle promoter elements are the sites for DREAM binding. These elements as well as the p53 response of Survivin, CDC25C, and PLK1 are evolutionarily conserved. No binding of p53 to these genes is detected by ChIP and mutation of proposed p53 binding sites does not alter the p53 response. Thus, a mechanism for direct p53-dependent transcriptional repression is not supported by the data. In contrast, repression by DREAM is consistent with most previous findings and unifies models based on p21-, E2F4-, p130-, and CDE/CHR-dependent repression by p53. In conclusion, the presented data suggest that the p53-p21-DREAM-CDE/CHR pathway regulates p53-dependent repression of Survivin, CDC25C, and PLK1