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

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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.

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Abbreviations

COX:

cyclooxygenase

PARP:

poly(ADP-ribose) polymerase

PG:

prostaglandin

R(+)-MA:

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

THC:

Δ9-Tetrahydrocannabinol

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Acknowledgment

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

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Authors and Affiliations

  1. Department of Experimental and Clinical Pharmacology and Toxicology, Friedrich Alexander University Erlangen–Nürnberg, Fahrstrasse 17, D-91054, Erlangen, Germany

    Karin Eichele, Ulrike Weinzierl, Robert Ramer, Kay Brune & Burkhard Hinz

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  1. Karin Eichele
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  2. Ulrike Weinzierl
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  3. Robert Ramer
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  4. Kay Brune
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  5. Burkhard Hinz
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Correspondence to Burkhard Hinz.

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Eichele, K., Weinzierl, U., Ramer, R. et al. R(+)-Methanandamide Elicits a Cyclooxygenase-2-Dependent Mitochondrial Apoptosis Signaling Pathway in Human Neuroglioma Cells. Pharm Res 23, 90–94 (2006). https://doi.org/10.1007/s11095-005-8815-2

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  • DOI: https://doi.org/10.1007/s11095-005-8815-2

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