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Akt regulates progesterone receptor B-dependent transcription and angiogenesis in endometrial cancer cells

Oncogene volume 35, pages 5191–5201 (2016)Cite this article

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Abstract

Progestins have long been used clinically for the treatment of endometrial cancers; however, the response rates to progestin therapy vary and the molecular mechanisms behind progestin insensitivity are poorly understood. We hypothesized that in PTEN-mutated endometrial cancers, hyperactive Akt signaling downregulates progesterone receptor B (PRB) transcriptional activity, leading to overall impaired progestin responses. We report that inhibition of Akt with the Akt inhibitor, MK-2206 (MK), in conjunction with progestin (R5020) treatment, is sufficient to upregulate a subset of PRB target genes in Ishikawa cells stably expressing PRB (PRB-Ishikawa). Through gene ontology analysis of Akt-regulated PRB target genes, angiogenesis was found to be the principle process regulated by Akt-PRB. To further interrogate the mechanism by which Akt modulates PRB transcriptional activity, ChIP-Mass spectrometry was performed to identify potential cofactors that differentially interact with PRB in the presence of R5020 and MK+R5020. 14-3-3σ was identified as a protein enriched in the MK+R5020 data set, and it was demonstrated that 14-3-3σ is required for the upregulation in PRB target gene expression following inhibition of Akt. To determine the ramifications of MK+R5020 treatment on angiogenesis, in vitro assays were performed and combinatorial MK+R5020 treatment significantly decreased endothelial cell invasion and tube formation more than MK or R5020 treatment alone. Furthermore, we found that combinatorial MK-2206+progesterone treatments decreased angiogenesis and proliferation in the Ptend/d conditional mouse model of endometrial cancer. Taken together, these findings suggest that a combinatorial therapeutic approach utilizing Akt inhibitors with progestins may improve the efficacy of progestin therapy for the treatment of endometrial cancer.

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Acknowledgements

We would like to acknowledge the Northwestern Genomics Core Facility, the Northwestern Proteomics Core Facility, and the Northwestern Next Generation Sequencing Core Facility. We would also like to thank members of the Kim laboratory for technical assistance and insightful discussion and review of the manuscript. This work is supported by NIH/NCI training grant T32CA09560 (IIL), Malkin Scholars Program from the Robert H. Lurie Comprehensive Cancer Center of Northwestern University (IIL), NIH/NICHD grant R01HD042311 (JPL) and NIH/NCI grant R01CA155513 (JJK).

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Authors and Affiliations

  1. Division of Reproductive Science in Medicine, Department of Obstetrics and Gynecology, Robert H. Lurie Comprehensive Cancer Center, Northwestern University Feinberg School of Medicine, Chicago, IL, USA

    I I Lee & J J Kim

  2. Department of Pathology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA

    K Maniar

  3. Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX, USA

    J P Lydon

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Correspondence to J J Kim.

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Lee, I., Maniar, K., Lydon, J. et al. Akt regulates progesterone receptor B-dependent transcription and angiogenesis in endometrial cancer cells. Oncogene 35, 5191–5201 (2016). https://doi.org/10.1038/onc.2016.56

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