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The antineoplastic agent α-bisabolol promotes cell death by inducing pores in mitochondria and lysosomes

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Abstract

The sesquiterpene α-bisabolol (α-BSB) has been shown to be an effective cytotoxic agent for a variety of human cancer cells in culture and animal models. However, much of its intracellular action remains elusive. We evaluated the cytotoxic action of α-BSB against CML-T1, Jurkat and HeLa cell lines, as preclinical models for myeloid, lymphoid and epithelial neoplasias. The approach included single cell analysis (flow cytometry, immunocytology) combined with cytotoxicity and proliferation assays to characterize organelle damage, autophagy, cytostatic effect, and apoptosis. The study focuses on the relevant steps in the cytotoxic cascade triggered by α-BSB: (1) the lipid rafts through which α-BSB enters the cells, (2) the opening of pores in the mitochondria and lysosomes, (3) the activation of both caspase-dependent and caspase-independent cell death pathways, (4) the induction of autophagy and (5) apoptosis. The effectiveness of α-BSB as an agent against tumor cells is grounded on its capability to act on different layers of cell regulation to elicit different concurrent death signals, thereby neutralizing a variety of aberrant survival mechanisms leading to treatment resistance in neoplastic cell.

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Abbreviations

AAF:

Autophagic activity factor

AO:

Acridine orange, a red fluorescent lysosomotropic probe

α-BSB:

α-bisabolol

AM:

Acetoxymethyl

CFSE:

Carboxyfluorescein succinimidyl ester

CM:

Complete medium

CM-H2DCFDA:

5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate, acetyl ester

CML-T1, Jurkat, HeLa cell lines:

Blast crisis of chronic myeloid leukemia, acute T cell leukemia, cervical cancer cell line, respectively

Cyto-ID® Green:

Cationic amphiphilic tracer that selectively stains autophagic vacuoles

DAPI:

4′,6-Diamidino-2-phenylindole, fluorescent nuclear counterstain

ΔΨm :

Mitochondrial transmembrane potential

FSC:

Forward scatter (in flow cytometry)

HBSS:

Hank’s balanced salt solution

IC50 :

Half maximal inhibitory concentration

JC-1:

5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide, red fluorescent mitochondrial counterstain

LT Green:

Lysosomotropic probe

mPTP:

Mitochondrial permeability transition pore

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

Q-VD-Oph:

Pan-caspase inhibitor

ROS:

Reactive oxygen species

SSC:

Side scatter (in flow cytometry)

TO-PRO®-3:

Red-fluorescent nuclear and chromosome counterstain

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Acknowledgments

FV wants to express his gratitude to Maria Langhieri for supporting cancer research in memory of her mother Iliana Tescaroli.

Authors’ contribution

FV conceived the research. AR performed the experiments. Both authors contributed to concept design, analyzed data, discussed results, wrote and approved the final manuscript.

Funding

This work was supported by funding from Italian Association for Cancer Research (AIRC, Milan, Italy)/Cariverona Foundation (Verona, Italy). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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  1. Section of Hematology, Cancer Research & Cell Biology Laboratory, Department of Medicine, University of Verona, Verona, Italy

    Antonella Rigo & Fabrizio Vinante

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  1. Antonella Rigo
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  2. Fabrizio Vinante
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Correspondence to Fabrizio Vinante.

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Rigo, A., Vinante, F. The antineoplastic agent α-bisabolol promotes cell death by inducing pores in mitochondria and lysosomes. Apoptosis 21, 917–927 (2016). https://doi.org/10.1007/s10495-016-1257-y

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