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Rgnef promotes ovarian tumor progression and confers protection from oxidative stress

Oncogene volume 38, pages 6323–6337 (2019)Cite this article

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Abstract

Ovarian cancer is the fifth-leading cause of cancer death among women. The dissemination of ovarian tumors and growth as spheroids accompanies late-stage disease. In cell culture, ovarian tumor cell spheroids can exhibit elevated resistance to environmental stressors, such as reactive oxygen species. Homeostatic balance of the antioxidant response is a protective mechanism that prevents anoikis, a form of programmed cell death. Signaling pathways activated by integrin receptors suppress anoikis. Rgnef (ARHGEF28/p190RhoGEF) is a guanine nucleotide exchange factor that is activated downstream of integrins. We find that Rgnef protein levels are elevated in late-stage serous ovarian cancer, high Rgnef mRNA levels are associated with decreased progression-free and overall survival, and genomic ARHGEF28 loss is associated with increased patient survival. Using transgenic and transplantable Rgnef knockout mouse models, we find that Rgnef is essential for supporting three-dimensional ovarian spheroid formation in vitro and tumor growth in mice. Using RNA-sequencing and bioinformatic analyses, we identify a conserved Rgnef-supported anti-oxidant gene signature including Gpx4, Nqo1, and Gsta4; common targets of the NF-kB transcription factor. Antioxidant treatment enhanced growth of Rgnef-knockout spheroids and Rgnef re-expression facilitated NF-κB-dependent tumorsphere survival. These studies reveal a new role for Rgnef in ovarian cancer to facilitate NF-κB-mediated gene expression protecting cells from oxidative stress.

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Acknowledgements

Supported by grants RO1CA180769, RO1CA102310, and UCSD Cancer Center Support grant P30CA023100, from UCSD Altman Clinical and Translational Research grant NIH UL1TR001442, and from charitable donations from Nine Girls Ask. DGS was supported by NIH RO1CA107263. KNT and AMB are fellows of the UCSD Reproductive Medicine Gynecologic Oncology Program and were supported by the Gaines Family fellowship. CDO was supported by NIH training grant (T32-CA121938). DCC was supported by NCI P30 CA006927, NCI CA195723, DOD W81XWH-16-1-0142 and charitable donations from The Roberta Dubrow Fund.

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  1. Nichol L. G. Miller

    Present address: Pfizer Inc., La Jolla, CA, 92121, USA

Authors and Affiliations

  1. Moores Cancer Center, Department of Obstetrics, Gynecology and Reproductive Science, UC San Diego Health, La Jolla, CA, 92093, USA

    Elizabeth G. Kleinschmidt, Nichol L. G. Miller, Duygu Ozmadenci, Isabelle Tancioni, Carlos Díaz Osterman, Allison M. Barrie, Kristin N. Taylor, Aaron Ye, Shulin Jiang, Dwayne G. Stupack & David D. Schlaepfer

  2. Biomedical Sciences Graduate Program, UC San Diego Health, La Jolla, CA, 92093, USA

    Elizabeth G. Kleinschmidt

  3. Fox Chase Cancer Center, Philadelphia, PA, 19111, USA

    Denise C. Connolly

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Contributions

Designing research studies: EGK, NLGM, DO, IT, CDO, DDS. Conducting experiments: EGK, NLGM, DO, IT, CDO, AMB, KNT, AY, SJ. Analyzing data: EGK, NLGM, DO, IT, CDO. Providing reagents: DCC, DGS. Writing the paper: EGK, NLGM, DCC, DGS, DDS. Study supervision: DCC, DGS, DDS

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Correspondence to David D. Schlaepfer.

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Kleinschmidt, E.G., Miller, N.L.G., Ozmadenci, D. et al. Rgnef promotes ovarian tumor progression and confers protection from oxidative stress. Oncogene 38, 6323–6337 (2019). https://doi.org/10.1038/s41388-019-0881-8

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