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In vitro siRNA-mediated GPX4 and AKT1 silencing in oxaliplatin resistance cancer cells induces ferroptosis and apoptosis

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Abstract

Oxaliplatin is a member of platinum-based chemotherapy drugs frequently used in colorectal cancer (CRC). However, resistance to oxaliplatin causes tumor progression and metastasis. Akt1 and Gpx4 are essential regulator genes of apoptosis and ferroptosis pathways. Inhibition of these genes might eradicate oxaliplatin resistance in resistant CRC cells. We compared two cell death strategies to reverse drug resistance in Caco-2 and HT-29 oxaliplatin-resistant cell lines. We used the AKT1-specific siRNA to induce apoptosis. Also, GPX4-specific siRNA and FIN56 were utilized to generate ferroptosis. The effect of these treatments was assessed by reactive oxygen species (ROS) formation, cell viability, and protein expression level assays. Besides, the expression of GPX4, CoQ10, and NRF2 was assessed in both cell lines after treatments. Correctly measuring the expression of these responsible genes and proteins confirms the occurrence of different types of cell death. In addition, the ability of Akt1/ GPX4 siRNA in resensitizing HT-29 and Caco-2 oxaliplatin resistance cells was evaluated. Our finding showed that the upregulation of GPX4/siRNA caused a reduction in GPX4 and CoQ10 expressions in both cell lines. However, the expression level of NRF2 showed the same level in our cell lines, so we observed a downregulation of NRF2 in resistant CRC cell lines. Cell viability assay indicated that induction of ferroptosis by GPX4/siRNA or FIN56 and apoptosis by Akt1/siRNA in resistant cell lines could reverse the oxaliplatin resistance. We concluded that downregulation of Akt1 or Gpx4 could increase the efficacy of oxaliplatin to overcome the resistance compared to FIN56.

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Funding

This project is part of a Ph.D. thesis (grant No. 59305) and Pharm.D. thesis (grant No.64435), funded by Drug Applied Research Center, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.

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Authors and Affiliations

  1. Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

    Morteza Golbashirzadeh, Hamid Reza Heidari, Ali Asghar Aghamolayi & Ahmad Yari Khosroushahi

  2. Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, P.O. 14766-51664, Tabriz, Iran

    Morteza Golbashirzadeh, Hamid Reza Heidari, Ali Asghar Aghamolayi & Yasin Fattahi

  3. Hematology and Oncology Research Center, Department of Applied Cell Sciences, School of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran

    Mehdi Talebi

  4. Department of Medical Nanotechnology, Faculty of Advanced Medical Science, Tabriz University of Medical Sciences, Daneshgah Street, P.O.Box 51548-53431, Tabriz, Iran

    Ahmad Yari Khosroushahi

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  1. Morteza Golbashirzadeh
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  2. Hamid Reza Heidari
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  3. Ali Asghar Aghamolayi
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Contributions

MG performed experiments, analyzed data, and wrote the paper. HRH supervised the research, analyzed the data, and prepared the manuscript; AAA performed experiments and analyzed the data. YF provided a resistance cell line and prepared a manuscript. MT served flow cytometry and analyzed its data. AYK designed the investigation and then led and supervised the project.

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Correspondence to Hamid Reza Heidari or Ahmad Yari Khosroushahi.

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Golbashirzadeh, M., Heidari, H.R., Aghamolayi, A.A. et al. In vitro siRNA-mediated GPX4 and AKT1 silencing in oxaliplatin resistance cancer cells induces ferroptosis and apoptosis. Med Oncol 40, 279 (2023). https://doi.org/10.1007/s12032-023-02130-6

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