Introduction/Background Endometrial cancer (EC) is the fourth most common cancer in women in the UK. While oestrogen and progesterone have been extensively researched, the role of androgen in EC has been relatively under-investigated. We aim to study the role of androgen in EC through the development of EC cell lines that overexpress androgen receptor (AR) with varying expression levels and investigate the consequences of AR overexpression on EC cell biology.

Methodology We transduced Ishikawa EC cell line with doxycycline-inducible ectopic AR, or constitutionally active ectopic AR in varying doses through lentivirus transduction resulting in varying levels of AR expression. This was followed by antibiotic selection and validation experiments to confirm ectopic AR expression. RNA was isolated from both cell models and RNA-sequencing was performed with and without androgen exposure to investigate the impact on the EC cells’ transcriptional profile.

Results Dose-dependent AR overexpression was observed using western plot, immunofluorescence, and qPCR, validating the expected post-transduction ectopic AR expression levels. RNA-seq analysis of the AR-transduced EC cells identified various differentially expressed genes in response to androgen exposure including CDHR1, KCNF1, COL8A1, and TIPARP. This effect on the transcriptional profile of EC cells was shown to be more pronounced with higher expression levels of AR. The differentially expressed genes were enriched for pathways associated with KRAS-signalling (nominal P-value=0.008), epithelial mesenchymal transition (nominal P-value<0.001), and apoptosis (nominal P-value=0.03) supporting the role of AR in EC pathogenesis.

Conclusion We successfully established AR-overexpressing EC cell lines which will serve as valuable models in investigating the AR role in EC. Our RNA-seq transcriptional profiling revealed pathways involved in tumorigenesis affected by AR expression highlighting the potential implications of AR signalling in EC. Future work will be performed to functionally validate the AR role in EC and ChIP-seq experiments will be conducted to identify direct AR target sites.

Disclosures -