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Impact of anthracycline-based chemotherapy on RB1 gene methylation in peripheral blood leukocytes and biomarkers of oxidative stress and inflammation in sarcoma patients

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Abstract

Purpose

We investigated the impact of anthracycline-based chemotherapy on methylation status of RB1 gene in peripheral blood leukocytes together with parameters of oxidative stress and inflammation in sarcoma patients.

Patients/methods

Blood samples were collected from 51 consecutive newly diagnosed sarcoma patients admitted to University Hospital Center Zagreb (Zagreb, Croatia) for first-line chemotherapy before the first cycle and post-chemotherapy. Methylation and copy number variation (CNV) of leukocyte RB1 gene were assessed using MS-MLPA probes. In addition, in blood samples, parameters of oxidative stress (ROS, MDA, SOD, and GSH) and inflammation (CRP, WBC, and NBC) were followed.

Results

In pre-chemotherapy samples, no CNVs and aberrant methylation of CpG106 promoter region of RB1 gene were detected; however, one patient had hypermethylation (by approximately 10%) of imprinted locus CpG85 in intron 2 of RB1 gene. In addition, a very good correlation of the tumor burden and CRP and tumor burden and GSH was found. The anthracycline-based chemotherapy reverts methylation of RB1 gene-imprinted locus CpG85 to normal level. Moreover, inflammation and oxidative stress parameters such as CRP, WBC, ROS, and MDA were significantly decreased in post-chemotherapy samples.

Conclusion

This single-centered study on a cohort of consecutive sarcoma patients indicates that sarcoma patients can have aberrant germline DNA methylation and confirms the relationship of tumor burden with inflammation and oxidative stress. The applied chemotherapy protocols reverted RB1 gene methylation to normal level and decreased the level of inflammation and oxidative damage, thus indicating chemotherapy benefit to the patient’s health status.

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Data availability

The data obtained in the study are presented within the article. Raw data are available upon request.

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Acknowledgements

The authors wish to thank Martina Demšer and staff of the Department of Oncology, University Hospital Center Zagreb, for organizing blood sampling and to Višnja Oreščanin for help with statistical analysis of data.

Funding

The study received financial support of University Hospital Center Zagreb and University of Zagreb (Z165). The study was supported by the project FarmInova (KK.01.1.1.02.0021) funded by the European Regional Development Fund.

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Authors and Affiliations

  1. Division of Cytogenetics, Department of Laboratory Diagnostics, University Hospital Centre Zagreb, Kišpatićeva 12, Zagreb, Croatia

    Anita Pokupec Bilić, Morana Mikloš & Ivana Tonković Đurišević

  2. Department of Pathophysiology, University of Zagreb School of Medicine, Šalata 2, Zagreb, Croatia

    Ivan Bilić

  3. Department of Oncology, University Hospital Centre Zagreb, Kišpatićeva 12, Zagreb, Croatia

    Ivan Bilić, Luka Simetić, Krešimir Blažičević & Davorin Herceg

  4. Department of Biology, University of Zagreb Faculty of Science, Ravnice 48, Zagreb, Croatia

    Sandra Radić Brkanac

  5. University of Zagreb Faculty of Pharmacy and Biochemistry, Kovačićeva 1, Zagreb, Croatia

    Ana-Marija Domijan

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  1. Anita Pokupec Bilić
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Contributions

Conceptualization: APB, IB, SRB, and AMD; methodology: APB, IB, SRB, and AMD; recruitment of patients and sample collection: IB, LS, KB, and DH; formal analysis and investigation: APB, MM, ITĐ, and AMD; writing––original draft preparation: APB, IB, and AMD; writing––review and editing: APB, IB, SRB, LS, KB, DH, and AMD. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Ana-Marija Domijan.

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The authors declare that they have no conflict of interest.

Ethical approval

The ethical approval was granted by the Ethics Committee of the University Hospital Center Zagreb (Zagreb, Croatia), approval no. 8.1-22/125-4 02/013 AG. The study observed the ethical principles of the Declaration of Helsinki.

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Pokupec Bilić, A., Bilić, I., Radić Brkanac, S. et al. Impact of anthracycline-based chemotherapy on RB1 gene methylation in peripheral blood leukocytes and biomarkers of oxidative stress and inflammation in sarcoma patients. Clin Transl Oncol 26, 1508–1518 (2024). https://doi.org/10.1007/s12094-023-03375-3

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