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Archives of Toxicology

, Volume 90, Issue 6, pp 1369–1382 | Cite as

Impact of a synthetic cannabinoid (CP-47,497-C8) on protein expression in human cells: evidence for induction of inflammation and DNA damage

  • Andrea Bileck
  • Franziska Ferk
  • Halh Al-Serori
  • Verena J. Koller
  • Besnik Muqaku
  • Alexander Haslberger
  • Volker Auwärter
  • Christopher Gerner
  • Siegfried Knasmüller
Molecular Toxicology

Abstract

Synthetic cannabinoids (SCs) are marketed worldwide as legal surrogates for marihuana. In order to predict potential health effects in consumers and to elucidate the underlying mechanisms of action, we investigated the impact of a representative of the cyclohexylphenols, CP47,497-C8, which binds to both cannabinoid receptors, on protein expression patterns, genomic stability and on induction of inflammatory cytokines in human lymphocytes. After treatment of the cells with the drug, we found pronounced up-regulation of a variety of enzymes in nuclear extracts which are involved in lipid metabolism and inflammatory signaling; some of the identified proteins are also involved in the endogenous synthesis of endocannabinoids. The assumption that the drug causes inflammation is further supported by results obtained in additional experiments with cytosols of LPS-stimulated lymphocytes which showed that the SC induces pro-inflammatory cytokines (IL12p40 and IL-6) as well as TNF-α. Furthermore, the proteome analyses revealed that the drug causes down-regulation of proteins which are involved in DNA repair. This observation provides an explanation for the formation of comets which was seen in single-cell gel electrophoresis assays and for the induction of micronuclei (which reflect structural and numerical chromosomal aberrations) by the drug. These effects were seen in experiments with human lymphocytes which were conducted under identical conditions as the proteome analysis. Taken together, the present findings indicate that the drug (and possibly other structurally related SCs) may cause DNA damage and inflammation in directly exposed cells of consumers.

Keywords

Synthetic cannabinoid Comet assay Lymphocytes Proteomics 

Abbreviations

B(a)P

Benzo(a)pyrene

BSA

Bovine serum albumin

CBs

Cannabinoid receptors

DDT

Dithiothreitol

IAA

Iodacetamide

FDR

False discovery rate

GC–MS

Gas chromatography–mass spectrometry

1H NMR

Proton nuclear magnetic resonance

LC–MS

Liquid chromatography–mass spectrometry

LFQ

Label-free quantification

MNi

Micronuclei

Nbuds

Nuclear buds

NDI

Nuclear division indices

NPBs

Nucleoplasmatic bridges

PAGE

Polyacrylamide gel electrophoresis

PBMCs

Peripheral blood mononuclear cells

PHA

Phytohemagglutinin

SCs

Synthetic cannabinoids

SCGE

Single-cell gel electrophoresis

SDS

Sodium dodecyl sulfate

THC

Tetrahydrocannabinol

Notes

Acknowledgments

This study was funded by the EU Commission in the frame of the coordinated Project “SPICE and synthetic cannabinoids” (JUST/2009/DPIP/AG/0948).

Compliance with ethical standards

Conflict of interest

The authors state that they have no conflict of interest.

Supplementary material

204_2015_1569_MOESM1_ESM.docx (11 kb)
Supplementary material 1 (DOCX 11 kb)
204_2015_1569_MOESM2_ESM.docx (90 kb)
Supplementary material 2 (DOCX 90 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Andrea Bileck
    • 1
  • Franziska Ferk
    • 2
  • Halh Al-Serori
    • 2
  • Verena J. Koller
    • 2
  • Besnik Muqaku
    • 1
  • Alexander Haslberger
    • 3
  • Volker Auwärter
    • 4
  • Christopher Gerner
    • 1
  • Siegfried Knasmüller
    • 2
  1. 1.Faculty of Chemistry, Institute of Analytical ChemistryUniversity of ViennaViennaAustria
  2. 2.Department of Internal Medicine 1, Comprehensive Cancer Center, Institute of Cancer ResearchMedical University of ViennaViennaAustria
  3. 3.Department of Nutritional SciencesUniversity of ViennaViennaAustria
  4. 4.Institute of Forensic MedicineUniversity Medical Center FreiburgFreiburgGermany

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