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β-Arrestin inhibition induces autophagy, apoptosis, G0/G1 cell cycle arrest in agonist-activated V2R receptor in breast cancer cells

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Abstract

Non-visual arrestins (β-arrestins) are endocytic proteins that mediate agonist-activated GPCRs internalization and signaling pathways in an independent manner. The involvement of β-arrestins in cancer invasion and metastasis is increasingly reported. So, it is hypothesized that inhibition of β-arrestins may diminish the survival chances of cancer cells. This study aimed to evaluate the in vitro impact of inhibiting β-arrestins on the autophagic and/or apoptotic responsiveness of breast cancer cells. We used Barbadin to selectively inhibit β-Arr/AP2 interaction in AVP-stimulated V2R receptor of triple-negative breast cancer cells (MDA MB-231). Autophagy was assessed by the microtubule-associated protein 1 light chain 3-II (LC3II), apoptosis was measured by Annexin-V/PI staining and cell cycle distribution was investigated based upon the DNA content using flow cytometry. Barbadin reduced cell viability to 69.1% and increased the autophagy marker LC3II and its autophagic effect disappeared in cells transiently starved in Earle's balanced salt solution (EBSS). Also, Barbadin mildly enhanced the expression of P62 mRNA and arrested 63.7% of cells in G0/G1 phase. In parallel, the drug-induced apoptosis in 29.9% of cells (by AV/PI) and 27.8% of cells were trapped in sub-G1 phase. The apoptotic effect of Barbadin was enhanced when autophagy was inhibited by the PI3K inhibitor (Wortmannin). Conclusively, the data demonstrate the dual autophagic and apoptotic effects of β-βArr/AP2 inhibition in triple-negative breast cancer cells. These observations nominate β-Arrs as selective targets in breast cancer treatment.

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Acknowledgement

Authors would like to express their gratitude toward Science and Technology Development Fund (STDF), for providing the financial supported to this work (grant number 34850).

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Authors and Affiliations

  1. Division of Biochemistry, Department of Chemistry, Faculty of Science, Tanta University, Tanta, Egypt

    Thoria Donia, Mohamed Abouda & Mohamed Hessien

  2. Department of Oncology, Faculty of Medicine, Ain Shams University, Cairo, Egypt

    Mohamed Kelany

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  1. Thoria Donia
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  2. Mohamed Abouda
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  3. Mohamed Kelany
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  4. Mohamed Hessien
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Contributions

Conceptualization: MH and TD; Methodology and investigations: MA, MK, TD and MH; Editing the original draft and revision: MH, MK and TD; Project administration, MH; Funding acquisition, MH, TD, MK.

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Correspondence to Mohamed Hessien.

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Donia, T., Abouda, M., Kelany, M. et al. β-Arrestin inhibition induces autophagy, apoptosis, G0/G1 cell cycle arrest in agonist-activated V2R receptor in breast cancer cells. Med Oncol 38, 38 (2021). https://doi.org/10.1007/s12032-021-01484-z

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