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The determination of the relationship between p97/VCP, small VCP-interacting protein, two ERAD proteins and steroidogenesis in Leydig cell lines

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Abstract

Background

In recent studies, it was shown that Endoplasmic reticulum-associated degradation (ERAD) is regulated by androgens and small VCP-interacting protein (SVIP) is an ERAD inhibitor. There is no data available about the interactions of ERAD proteins with proteins involved in steroidogenesis. The aim of the study was to investigate the expressions of SVIP, p97/VCP, StAR, CYP17A1 and 3β-HSD in human and mouse.

Methods and results

HLC, TM3 and MA-10 Leydig cell lines were used to determine roles of ERAD proteins in steroidogenesis based on immunofluorescence, Western blot, qRT-PCR, ELISA. Findings showed that StAR, CYP17A1 and 3β-HSD were colocalized with SVIP and p97/VCP in Leydig cells. A decrease in CYP17A1, 3β-HSD and StAR expressions was observed as a result of suppression of SVIP siRNAs and p97/VCP siRNAs expressions in MA10, TM3 and HLC. When siSVIP transfected cells were compared with siSVIP transfected with hCG-exposed cells, SVIP protein expression was significantly increased as compared to the SVIP transfected group in human Leydig cells.

Conclusion

We suggest that the suppression of protein expressions by p97/VCP and SVIP siRNAs in Leydig cells, the effects of proteins involved in steroidogenesis (StAR, CYP17A1 and 3β-HSD) have proven to be originating from p97/VCP and SVIP which were playing a role in the steroidogenesis process. Additionally, it was demonstrated that testosterone levels decreased after transfection with p97/VCP siRNA and SVIP siRNA, p97/VCP and SVIP created an effect on testosterone synthesis while taking place in the steps of testosterone synthesis. Further, it was determined in the study that the SVIP was affected by hCG stimulations.

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Knowledge and Acknowledgment

Part of this work was supported by The Scientific and Technological Research Council of Turkey (TUBITAK), Grant ID: 119S249.

Funding

Part of this work was supported by The Scientific and Technological Research Council of Turkey (TUBITAK), Grant ID: 119S249.

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Authors and Affiliations

  1. Medical Faculty, Department of Histology and Embryology Ankara, Ankara Yıldırım Beyazıt University, Ankara, Turkey

    Ebru Alimogullari, Gülben Akcan & Sevil Cayli

  2. Central Research and Application Center, Ankara Yıldırım Beyazıt University, Ankara, Turkey

    Oguz Ari

Authors
  1. Ebru Alimogullari
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  2. Gülben Akcan
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  3. Oguz Ari
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  4. Sevil Cayli
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Contributions

Ebru Alimogullari was involved in the project design, project development and manuscript writing. Sevil Cayli, Gulben Akcan, and Oguz Ari contributed to project development, data collection, and data analysis

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Correspondence to Ebru Alimogullari.

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Alimogullari, E., Akcan, G., Ari, O. et al. The determination of the relationship between p97/VCP, small VCP-interacting protein, two ERAD proteins and steroidogenesis in Leydig cell lines. Mol Biol Rep 49, 9159–9170 (2022). https://doi.org/10.1007/s11033-022-07740-3

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  • DOI: https://doi.org/10.1007/s11033-022-07740-3

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