Proteomic analysis of stromal and epithelial cell communications in human endometrial cancer using a unique 3D co-culture model.

Epithelial and stromal communications are essential for normal uterine functions and their dysregulation contributes to the pathogenesis of many diseases including infertility, endometriosis, and cancer. Although many studies have highlighted the advantages of culturing cells in three-dimensional (3D) compared to the conventional two-dimensional (2D) culture system, one of the major limitations of these systems is the lack of incorporation of cells from non-epithelial lineages. In an effort to develop a culture system incorporating both stromal and epithelial cells, we developed 3D endometrial cancer spheroids by co-culturing endometrial stromal cells with cancerous epithelial cells. The spheroids developed by this method were phenotypically comparable to in vivo endometrial cancer tissue. Proteomic analysis of our co-culture spheroids comparable to human endometrial tissue revealed 591 common proteins and canonical pathways that are closely related to endometrium biology. To determine the feasibility of using this model for drug screening, we tested the efficacy of tamoxifen and everolimus. In summary, we have developed a unique 3D model system of human endometrial cancer that will serve as the foundation for the further development of 3D culture systems incorporating different cell types of the human uterus for deciphering the contributions of non-epithelial cells present in cancer microenvironment. This article is protected by copyright. All rights reserved.