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Pharmaceutical Research

, Volume 23, Issue 1, pp 90–94 | Cite as

R(+)-Methanandamide Elicits a Cyclooxygenase-2-Dependent Mitochondrial Apoptosis Signaling Pathway in Human Neuroglioma Cells

  • Karin Eichele
  • Ulrike Weinzierl
  • Robert Ramer
  • Kay Brune
  • Burkhard HinzEmail author
Research Paper

Purpose

Cannabinoids have been associated with tumor regression and apoptosis of cancer cells. Recently, we have shown that R(+)-methanandamide (R(+)-MA) induces apoptosis of H4 human neuroglioma cells via a mechanism involving de novo expression of the cyclooxygenase-2 (COX-2) enzyme. The present study investigated a possible involvement of a mitochondrial-driven pathway in this process.

Methods

Cell death was determined by the WST-1 cell viability test, and changes in apoptotic parameters [i.e., release of mitochondrial cytochrome c, activation of caspases, cleavage of poly(ADP-ribose) polymerase (PARP)] were detected by Western blotting.

Results

H4 cells treated with R(+)-MA showed typical signs of mitochondrial apoptosis, i.e., release of mitochondrial cytochrome c into the cytosol and activation of initiator caspase-9. Moreover, activation of the executor caspase-3 was observed following cannabinoid treatment. Cells were fully protected from apoptotic cell death by the caspase-3 inhibitor Ac-DEVD-CHO, indicating a crucial role for caspase-3 activation in R(+)-MA-elicited apoptosis. Furthermore, cleavage of the caspase-3 target protein PARP was registered. All of the aforementioned effects were substantially reduced by the selective COX-2 inhibitor celecoxib (1 μM) at a pharmacologically relevant, nonapoptotic concentration.

Conclusion

R(+)-MA-induced apoptosis is mediated via a mitochondrial-dependent pathway that becomes activated, at least in part, through up-regulation of the COX-2 enzyme.

Key Words

cyclooxygenase-2 human neuroglioma cells intrinsic apoptosis mitochondrial apoptosis R(+)-methanandamide 

Abbreviations

COX

cyclooxygenase

PARP

poly(ADP-ribose) polymerase

PG

prostaglandin

R(+)-MA

R(+)-methanandamide [R-(+)-arachidonyl-1′-hydroxy-2′-propylamide)]

THC

Δ9-Tetrahydrocannabinol

Notes

Acknowledgment

This study was supported by the Deutsche Forschungsgemeinschaft (HI 813/1-1 and SFB 539 TP BI.6).

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Copyright information

© Springer Science + Business Media, Inc. 2006

Authors and Affiliations

  • Karin Eichele
    • 1
  • Ulrike Weinzierl
    • 1
  • Robert Ramer
    • 1
  • Kay Brune
    • 1
  • Burkhard Hinz
    • 1
    Email author
  1. 1.Department of Experimental and Clinical Pharmacology and ToxicologyFriedrich Alexander University Erlangen–NürnbergErlangenGermany

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