Next Generation Sequencing of Ovarian Cancer Cells upon Exposure to Ascites from Wild Type and TG2-/- Mice

The goal of this study was to compare signaling pathways in ovarian cancer cells exposed in vitro or in vivo to peritoneal tumor microenvironment of host TG2+/+ vs. TG2-/- mice. For this, we performed NGS-derived transcriptome profiling (RNA-seq) followed by Ingenuity Pathway Analysis (IPA) of ID8 ovarian cancer cells. Overall design: Two approaches were undertaken: 1. ID8 ovarian cancer cells were treated in vitro for 5 days with ascites collected from wild type (WT) or TG2-/- mice; 2. ID8 cells were injected intraperitoneally in WT and TG2-/- mice. At the end of tumor growth experiment (day 45), ovarian cancer cells (EpCAM+) were recovered by magnetic cell separation. mRNA profiles were generated by deep sequencing, in duplicate (approach 1) or triplicate (approach 2), using Illumina technology.