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Ribosomal S6 kinase 4 (RSK4) tumor suppressor gene promoter methylation status in ovarian cancer

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Abstract

Background

Previously, we reported lower RSK4 mRNA and protein levels in malignant ovarian tumors compared to normal and benign ovarian tissues. Also, we observed a significant inverse correlation between the advanced ovarian cancer stages and RSK4 mRNA levels. We did not investigate the mechanisms involved in RSK4-reduced expression in ovarian cancer. Thus, this study investigates whether RSK4 promoter methylation in ovarian cancer tissues is responsible for its low expression. Additionally, the reactivation of RSK4 expression and its effect was studied in ovarian cancer cell lines.

Methods and results

RSK4 promoter methylation percentage in malignant and benign ovarian tumors and normal ovary tissues was determined by combined bisulfite restriction analysis. The reactivation of RSK4 expression by decitabine treatment was studied in OVCAR3, SKOV3, TOV-112D, and TOV-21G cells by Western blotting. Cell proliferation was determined by XTT. A significantly high methylation percentage of the RSK4 promoter was observed among malignant and benign ovarian tumors but not in normal ovarian tissue. RSK4 promoter methylation was not associated with age, histological subtype, or stages of ovarian cancer. RSK4 promoter methylation correlates weakly but not significantly with RSK4 protein expression. No correlation was shown between RSK4 methylation and RSK4 mRNA expression. Decitabine induces RSK4 reactivation in all cell lines. However, cell proliferation was reduced only in TOV-112D cells.

Conclusion

These data indicate that although RSK4 promoter methylation is increased in malignant ovarian tumors, this mechanism is unlikely to regulate its expression in ovarian cancer. RSK4 reactivation reduced cell proliferation only in the endometroid histological subtype.

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Data availability

The data that support the findings of this study are available from the corresponding author, Laura Diaz-Cueto, MD., PhD., upon reasonable request.

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Funding

This study was supported by a Grant (FIS/IMSS/PROT/G12/1142) to L. Diaz-Cueto from the Coordinación de Investigación en Salud. Fondo de Investigación en Salud, Instituto Mexicano del Seguro Social. Ciudad de México. México.

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Authors and Affiliations

  1. Unidad de Investigación Médica en Medicina Reproductiva, UMAE Hospital de Gineco-Obstetricia No. 4 “Luis Castelazo Ayala”, Instituto Mexicano del Seguro Social, Av. Rio Magdalena No. 289, Sexto piso. Tizapán, San Angel, CP 01090, Mexico City, Mexico

    Fabian Arechavaleta-Velasco, Pablo Dominguez-Lopez, Ernesto Acosta-Jimenez & Laura Diaz-Cueto

  2. Hospital Angeles del Pedregal, Mexico City, Mexico

    Ernesto Acosta-Jimenez

Authors
  1. Fabian Arechavaleta-Velasco
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  2. Pablo Dominguez-Lopez
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  3. Ernesto Acosta-Jimenez
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  4. Laura Diaz-Cueto
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Contributions

All authors listed have contributed substantially and approved the manuscript. LDC conceived the original idea, designed the experiments, and supervised the project. PDL and EAJ performed all the experiments. LDC and FAV analyzed the data and wrote the paper.

Corresponding author

Correspondence to Laura Diaz-Cueto.

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Competing interest

The authors declare that they have no competing interests.

Ethical approval

The UMAE Hospital de Gineco-Obstetricia No. 4 Dr. Luis Castelazo Ayala institutional review board approved this study (R-2012-3606-1).

Consent to participate

Ovarian tissue samples were collected from our tissue bank. Previously, all patients gave written informed consent for the retention and analysis of their tissue for research purposes.

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Not applicable.

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Arechavaleta-Velasco, F., Dominguez-Lopez, P., Acosta-Jimenez, E. et al. Ribosomal S6 kinase 4 (RSK4) tumor suppressor gene promoter methylation status in ovarian cancer. Mol Biol Rep 50, 6863–6870 (2023). https://doi.org/10.1007/s11033-023-08609-9

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  • DOI: https://doi.org/10.1007/s11033-023-08609-9

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