Cited by 33Flavonoids in Medical ResearchNeuroblastoma Research and TreatmentsPharmacological Effects of Natural Compounds

Anti-proliferative and cytotoxic activities of the flavonoid isoliquiritigenin in the human neuroblastoma cell line SH-SY5Y

Escobar, Stephane J.de M., Fong, Genevieve M., Winnischofer, Sheila M.B., Simone, Martin, Munoz, Lenka, Dennis, Joanne M., Rocha, Maria Eliane M., Witting, Paul K.·2019·Chemico-Biological Interactions

Abstract

Neuroblastoma is a common childhood cancer with high mortality. We evaluated the capacity of the flavonoid, isoliquiritigenin (4,2',4'-trihydroxychalcone; ISL) to inhibit cellular proliferation and migration in the human neuroblastoma cell line SH-SY5Y. Incubation of cultured SH-SY5Y cells with 20-100 μM ISL decreased cell confluency (15-70%) after 24 h incubation, while 10-100 μM ISL (24 h) depleted intracellular ATP stores (15-90% vs vehicle-treated control) after 24 h incubation. ISL-mediated cell toxicity did not involve intracellular caspase 3/7 activation, externalization of phosphatidylserine on the cell membrane or stimulation of TNF and IL-1β release, all indicating that the flavonoid did not induce apoptosis. Pre-treatment of cells with necrostatin-1, a necroptosis inhibitor, significantly restored ATP levels (ATP levels increased 12-42%) in ISL-treated neuroblastoma cells indicative of enhanced viability. By contrast, RIP1 phosphorylation status remained unchanged in cells treated with ISL although the intracellular ratio of phosphorylated/total parental RIP1 increased after ISL treatment on SH-SY5Y cells indicating that ISL decreased levels of native RIP1. In addition, ISL treatment inhibited SH-SY5Y cell migration/proliferation in a scratch assay and arrested cell cycle transition by significantly decreasing the number of cells in G0/G1 phase and increasing populations by ~10% in S (primarily) and G2/M (lesser extent) phases. The intracellular ratio of phosphorylated/total ERK 1/2 and p38 remained unchanged after ISL treatment (up to 40 μM); ERK activation was only determined at ISL dose well above the experimental IC 50 value as judged by ELISA analyses and this did not correlate with ISL cytotoxicity at lower dose <40 μM; Western blot assay confirmed the detection of phosphorylated (p-)ERK1/2 and (p-)p38 in ISL treated cells. Together the results suggest that ISL exerts anti-proliferative and cytotoxic activity on SHSY5Y cells through the loss of ATP, induction of cell cycle arrest, and cell death largely via a necroptotic mechanism in the absence of apoptotic activity.

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Crossref · 10.1016/j.cbi.2018.11.022

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Authors
Escobar, S. J. D. M., Fong, G. M., Winnischofer, S. M. B., Simone, M., Munoz, L., Dennis, J. M., Rocha, M. E. M., & Witting, P. K.
Affiliations
  • The University of Sydney (AU)
  • Universidade Federal do Paraná (BR)
Corresponding author
Witting, Paul K.
Venue
Chemico-Biological Interactions
Publisher
Elsevier BV
Published
2019-02
Volume
299
Pages
77-87
ISSN
0009-2797 (ISSN-L)
OpenAlex
W2903134099
Cited by
33
References
73
Citations by year
2026: 2 · 2025: 5 · 2024: 3 · 2023: 8 · 2022: 4 · 2021: 4 · 2020: 5 · 2019: 2
FWCI
1.78
Type
journal article
Language
en
Open access
closed
Locations
  • Chemico-Biological Interactions (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Australian Research Council (DP130103711, DP160102063)
  • Sydney Medical School
  • The University of Sydney
  • Brazilian Science Exchange Scholarship
Topics
  • Health Sciences › Medicine › Pharmacology › Flavonoids in Medical Research
  • Health Sciences › Medicine › Neurology › Neuroblastoma Research and Treatments
  • Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacological Effects of Natural Compounds
Keywords
isoliquiritigenin, flavonoids, neuroblastoma, SH-SY5Y cells, antiproliferative activity, cytotoxicity, necroptosis, ATP depletion, cell cycle arrest, ERK1/2, p38 MAPK, RIPK1, caspase 3/7, apoptosis, cell migration, chalcones, anticancer agents
MeSH terms
Adenosine Triphosphate, Cell Membrane Permeability, Cell Movement, Flavonoids, Humans, Imidazoles, Indoles, Microtubule-Associated Proteins, Neuroblastoma, Phosphorylation, Tumor Necrosis Factor-alpha, Mitogen-Activated Protein Kinase 1, Cell Line, Tumor, Chalcones, Mitogen-Activated Protein Kinase 3, Cell Proliferation, Caspase 3, Interleukin-1beta, G1 Phase Cell Cycle Checkpoints

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