Modeling effects of diabetes and obesity co-morbidities in endometrial cancer development and progression

Abstract

Methods The effects of insulin and insulin like growth factors 1 and 2 (IGF1 and 2) on proliferation were measured in primary cultures of human endometrial cells using a cytotoxicity assay, while transformation was measured by soft agar colony formation and histologic evaluation of and organotypic model of endometrial carcinogenesis and chemoprevetion [1]. Akt phosphorylation status was measured by Western blot. Levels of leptin, adiponectin adipsin, c-peptide, tumor necrosis factor a (TNFa) chemokine (C-C motif) ligand 2 (CCL2, also called monocyte chemotactic protein-1 or MCP-1), Interleukin (IL)-1b, IL10, vascular endothelial growth factor (VEGF) and insulin in serum from 84 endometrial cancer patients were measured using multiplex magnetic bead assays (BioRad and Millipore). Glucose was measured using a kit (Cayman), and HOMA Scores were calculated. These biomarkers were compared with tumor stage, grade, recurrence of disease and age at diagnosis using Spearman Correlation. Results Insulin, IGF1 and IGF2 caused dose-responsive induction of endometrial cell proliferation, which surprisingly was associated with loss of Akt phosphorylation on Ser473. Insulin treatment as a single agent induced anchorageindependent colony formation in the soft agar assay and histologic features of transformed cells in organotypic cultures. Insulin also enhanced these transformation measures induced by 7,12-Dimethylbenz(a)anthracene (DMBA) carcinogen. Elevated CCL2 in serum of endometrial cancer patients was significantly associated with worse stage of disease (r=0.262, p=0.018). Reduced serum TNFa was significantly associated with increased age at diagnosis (r=0.332, p=0.048). No other associations between biomarkers and patient characteristics were noted.