Cytotoxicity of ungeremine towards multi-factorial drug resistant cancer cells and induction of apoptosis, ferroptosis, necroptosis and autophagy

Armelle T Mbaveng; Gabin T M Bitchagno; Victor Kuete; Pierre Tane; Thomas Efferth

doi:10.1016/j.phymed.2019.152832

Cytotoxicity of ungeremine towards multi-factorial drug resistant cancer cells and induction of apoptosis, ferroptosis, necroptosis and autophagy

Phytomedicine. 2019 Jul:60:152832. doi: 10.1016/j.phymed.2019.152832. Epub 2019 Jan 15.

Authors

Armelle T Mbaveng¹, Gabin T M Bitchagno², Victor Kuete¹, Pierre Tane², Thomas Efferth³

Affiliations

¹ Department of Pharmaceutical Biology, Institute of Pharmacy and Biochemistry, University of Mainz, Staudinger Weg 5, 55128 Mainz, Germany; Department of Biochemistry, Faculty of Science, University of Dschang, P.O. Box 67, Dschang, Cameroon.
² Department of Chemistry, Faculty of Science, University of Dschang, P.O. Box 67, Dschang, Cameroon.
³ Department of Pharmaceutical Biology, Institute of Pharmacy and Biochemistry, University of Mainz, Staudinger Weg 5, 55128 Mainz, Germany. Electronic address: efferth@uni-mainz.de.

PMID: 31031043
DOI: 10.1016/j.phymed.2019.152832

Abstract

Background: Successful cancer chemotherapy is hampered by resistance of cancer cells to established anticancer drugs. Numerous natural products reveal cytotoxicity towards tumor cells.

Purpose: The present study was aimed to determine the cytotoxicity of a betaine-type alkaloid, ungeremine, towards 9 cancer cell lines including various sensitive and drug-resistant phenotypes. The mode of action of this compound was further investigated.

Methods: The cytotoxicity, ferroptotic and necroptotic cell death were determined by the resazurin reduction assay. Caspase activation was evaluated using the caspase-Glo assay. Flow cytometry was applied for the analysis of cell cycle analysis (PI staining), apoptosis (annexin V/PI staining), mitochondrial membrane potential (MMP) (JC-1) and reactive oxygen species (ROS) (H₂DCFH-DA). Apoptotic, necroptotic and autophagic markers were determined by Western blotting. CCRF-CEM leukemia cells were used for all mechanistic studies.

Results: Ungeremine displayed cytotoxic activity towards the 9 cancer cell lines tested, including drug-sensitive and MDR phenotypes. The IC₅₀values obtained varied from 3.67 µM (in MDA-MB-231-BCRP breast carcinoma cells) to 75.24 µM (against in CEM/ADR5000 leukemia cells) for ungeremine and from 0.02 µM (against CCRF-CEM cells) to 122.96 µM (against CEM/ADR5000 cells) for doxorubicin (control drug). Ungeremine induced ferroptosis, necroptosis, autophagy as well as apoptosis mediated by caspase activation, MMP alteration and increase ROS production.

Conclusion: The present investigation showed that ungeremine is a promising cytotoxic compoundthat could be further explored in the future to develop new anticancer drugs to fight sensitive and resistant phenotypes.

Keywords: Alkaloid; Cell death; Multi-drug resistance; Natural product.

MeSH terms

Alkaloids / toxicity*
Amaryllidaceae Alkaloids / chemistry
Amaryllidaceae Alkaloids / toxicity*
Antineoplastic Agents, Phytogenic / chemistry
Antineoplastic Agents, Phytogenic / toxicity*
Apoptosis / drug effects*
Autophagy / drug effects*
Cell Death / drug effects
Cell Line, Tumor
Drug Resistance, Multiple / drug effects
Drug Resistance, Neoplasm / drug effects
Humans
Indolizines / chemistry
Indolizines / toxicity*
Plant Extracts / chemistry
Plant Extracts / toxicity*
Reactive Oxygen Species / metabolism

Substances

Alkaloids
Amaryllidaceae Alkaloids
Antineoplastic Agents, Phytogenic
Indolizines
Plant Extracts
Reactive Oxygen Species
ungeremine