Arsenic trioxide (As2O3) induced calcium signals and cytotoxicity in two human cell lines: SY-5Y neuroblastoma and 293 embryonic kidney (HEK)

Ana Maria Florea, Frank Splettstoesser, Dietrich Busselberg

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

Arsenic trioxide (As2O3) has anticancer properties; however, its use also leads to neuro-, hepato- or nephro-toxicity, and therefore, it is important to understand the mechanism of As2O3 toxicity. We studied As2O3 influence on intracellular calcium ([Ca2+]i) homeostasis of human neuroblastoma SY-5Y and embryonic kidney cells (HEK 293).We also relate the As2O3 induced [Ca2+]i modifications with cytotoxicity. We used Ca2+ sensitive dyes (fluo-4 and rhod-2) combined with laser scanning microscopy or fluorescence activated cell sorting to measure Ca2+ changes during the application of As2O3 and we approach evaluation of cytotoxicity. As2O3 (1 μM) increased [Ca2+]i in SY-5Y and HEK 293 cells. Three forms of [Ca2+]i-elevations were found: (1) steady-state increases, (2) transient [Ca2+]i-elevations and (3) Ca2+-spikes. [Ca2+]i modifications were independent from extracellular Ca2+ but dependent on internal calcium stores. The effect was not reversible. Inositol triphosphate (IP3) and ryanodine (Ry) receptors are involved in regulation of signals induced by As2O3. 2-APB and dantrolene significantly reduced the [Ca2+]i-rise (p < 0.001, t-test) but did not completely abolish [Ca2+]i-elevation or spiking. This indicates that other Ca2+ regulating mechanisms are involved. In cytotoxicity tests As2O3 significantly reduced cell viability in both cell types. Staining with Hoechst 33342 showed occurrence of apoptosis and DNA damage. Our data suggest that [Ca2+]i is an important messenger in As2O3 induced cell death.

Original languageEnglish
Pages (from-to)292-301
Number of pages10
JournalToxicology and Applied Pharmacology
Volume220
Issue number3
DOIs
Publication statusPublished - 1 May 2007
Externally publishedYes

Fingerprint

Cytotoxicity
Neuroblastoma
Cells
Calcium
Kidney
Cell Line
Toxicity
Inositol 1,4,5-Trisphosphate Receptors
HEK293 Cells
arsenic trioxide
Dantrolene
Ryanodine Receptor Calcium Release Channel
Inositol
Cell death
Sorting
Confocal Microscopy
DNA Damage
Cell Survival
Flow Cytometry
Microscopic examination

Keywords

  • Apoptosis
  • Arsenic trioxide
  • AsO
  • Calcium homeostasis
  • Calcium signals
  • HEK
  • IP receptors
  • Neuroblastoma
  • Ryanodine receptors
  • Tumor cells

ASJC Scopus subject areas

  • Toxicology
  • Pharmacology

Cite this

Arsenic trioxide (As2O3) induced calcium signals and cytotoxicity in two human cell lines : SY-5Y neuroblastoma and 293 embryonic kidney (HEK). / Florea, Ana Maria; Splettstoesser, Frank; Busselberg, Dietrich.

In: Toxicology and Applied Pharmacology, Vol. 220, No. 3, 01.05.2007, p. 292-301.

Research output: Contribution to journalArticle

@article{bf733c0196ea41cd9f8624f580e8d0a0,
title = "Arsenic trioxide (As2O3) induced calcium signals and cytotoxicity in two human cell lines: SY-5Y neuroblastoma and 293 embryonic kidney (HEK)",
abstract = "Arsenic trioxide (As2O3) has anticancer properties; however, its use also leads to neuro-, hepato- or nephro-toxicity, and therefore, it is important to understand the mechanism of As2O3 toxicity. We studied As2O3 influence on intracellular calcium ([Ca2+]i) homeostasis of human neuroblastoma SY-5Y and embryonic kidney cells (HEK 293).We also relate the As2O3 induced [Ca2+]i modifications with cytotoxicity. We used Ca2+ sensitive dyes (fluo-4 and rhod-2) combined with laser scanning microscopy or fluorescence activated cell sorting to measure Ca2+ changes during the application of As2O3 and we approach evaluation of cytotoxicity. As2O3 (1 μM) increased [Ca2+]i in SY-5Y and HEK 293 cells. Three forms of [Ca2+]i-elevations were found: (1) steady-state increases, (2) transient [Ca2+]i-elevations and (3) Ca2+-spikes. [Ca2+]i modifications were independent from extracellular Ca2+ but dependent on internal calcium stores. The effect was not reversible. Inositol triphosphate (IP3) and ryanodine (Ry) receptors are involved in regulation of signals induced by As2O3. 2-APB and dantrolene significantly reduced the [Ca2+]i-rise (p < 0.001, t-test) but did not completely abolish [Ca2+]i-elevation or spiking. This indicates that other Ca2+ regulating mechanisms are involved. In cytotoxicity tests As2O3 significantly reduced cell viability in both cell types. Staining with Hoechst 33342 showed occurrence of apoptosis and DNA damage. Our data suggest that [Ca2+]i is an important messenger in As2O3 induced cell death.",
keywords = "Apoptosis, Arsenic trioxide, AsO, Calcium homeostasis, Calcium signals, HEK, IP receptors, Neuroblastoma, Ryanodine receptors, Tumor cells",
author = "Florea, {Ana Maria} and Frank Splettstoesser and Dietrich Busselberg",
year = "2007",
month = "5",
day = "1",
doi = "10.1016/j.taap.2007.01.022",
language = "English",
volume = "220",
pages = "292--301",
journal = "Toxicology and Applied Pharmacology",
issn = "0041-008X",
publisher = "Academic Press Inc.",
number = "3",

}

TY - JOUR

T1 - Arsenic trioxide (As2O3) induced calcium signals and cytotoxicity in two human cell lines

T2 - SY-5Y neuroblastoma and 293 embryonic kidney (HEK)

AU - Florea, Ana Maria

AU - Splettstoesser, Frank

AU - Busselberg, Dietrich

PY - 2007/5/1

Y1 - 2007/5/1

N2 - Arsenic trioxide (As2O3) has anticancer properties; however, its use also leads to neuro-, hepato- or nephro-toxicity, and therefore, it is important to understand the mechanism of As2O3 toxicity. We studied As2O3 influence on intracellular calcium ([Ca2+]i) homeostasis of human neuroblastoma SY-5Y and embryonic kidney cells (HEK 293).We also relate the As2O3 induced [Ca2+]i modifications with cytotoxicity. We used Ca2+ sensitive dyes (fluo-4 and rhod-2) combined with laser scanning microscopy or fluorescence activated cell sorting to measure Ca2+ changes during the application of As2O3 and we approach evaluation of cytotoxicity. As2O3 (1 μM) increased [Ca2+]i in SY-5Y and HEK 293 cells. Three forms of [Ca2+]i-elevations were found: (1) steady-state increases, (2) transient [Ca2+]i-elevations and (3) Ca2+-spikes. [Ca2+]i modifications were independent from extracellular Ca2+ but dependent on internal calcium stores. The effect was not reversible. Inositol triphosphate (IP3) and ryanodine (Ry) receptors are involved in regulation of signals induced by As2O3. 2-APB and dantrolene significantly reduced the [Ca2+]i-rise (p < 0.001, t-test) but did not completely abolish [Ca2+]i-elevation or spiking. This indicates that other Ca2+ regulating mechanisms are involved. In cytotoxicity tests As2O3 significantly reduced cell viability in both cell types. Staining with Hoechst 33342 showed occurrence of apoptosis and DNA damage. Our data suggest that [Ca2+]i is an important messenger in As2O3 induced cell death.

AB - Arsenic trioxide (As2O3) has anticancer properties; however, its use also leads to neuro-, hepato- or nephro-toxicity, and therefore, it is important to understand the mechanism of As2O3 toxicity. We studied As2O3 influence on intracellular calcium ([Ca2+]i) homeostasis of human neuroblastoma SY-5Y and embryonic kidney cells (HEK 293).We also relate the As2O3 induced [Ca2+]i modifications with cytotoxicity. We used Ca2+ sensitive dyes (fluo-4 and rhod-2) combined with laser scanning microscopy or fluorescence activated cell sorting to measure Ca2+ changes during the application of As2O3 and we approach evaluation of cytotoxicity. As2O3 (1 μM) increased [Ca2+]i in SY-5Y and HEK 293 cells. Three forms of [Ca2+]i-elevations were found: (1) steady-state increases, (2) transient [Ca2+]i-elevations and (3) Ca2+-spikes. [Ca2+]i modifications were independent from extracellular Ca2+ but dependent on internal calcium stores. The effect was not reversible. Inositol triphosphate (IP3) and ryanodine (Ry) receptors are involved in regulation of signals induced by As2O3. 2-APB and dantrolene significantly reduced the [Ca2+]i-rise (p < 0.001, t-test) but did not completely abolish [Ca2+]i-elevation or spiking. This indicates that other Ca2+ regulating mechanisms are involved. In cytotoxicity tests As2O3 significantly reduced cell viability in both cell types. Staining with Hoechst 33342 showed occurrence of apoptosis and DNA damage. Our data suggest that [Ca2+]i is an important messenger in As2O3 induced cell death.

KW - Apoptosis

KW - Arsenic trioxide

KW - AsO

KW - Calcium homeostasis

KW - Calcium signals

KW - HEK

KW - IP receptors

KW - Neuroblastoma

KW - Ryanodine receptors

KW - Tumor cells

UR - http://www.scopus.com/inward/record.url?scp=34247245667&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34247245667&partnerID=8YFLogxK

U2 - 10.1016/j.taap.2007.01.022

DO - 10.1016/j.taap.2007.01.022

M3 - Article

C2 - 17376498

AN - SCOPUS:34247245667

VL - 220

SP - 292

EP - 301

JO - Toxicology and Applied Pharmacology

JF - Toxicology and Applied Pharmacology

SN - 0041-008X

IS - 3

ER -