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Mevalonate Prevents Lovastatin-Induced Apoptosis in Medulloblastoma Cell Lines

Published online by Cambridge University Press:  02 December 2014

Wei Wang  and
Robert J.B. Macaulay
Wei Wang
Affiliation:
Department of Pathology, College of Medicine, University of Saskatchewan, Saskatoon, SK, Canada
Robert J.B. Macaulay
Affiliation:
Department of Pathology, College of Medicine, University of Saskatchewan, Saskatoon, SK, Canada
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Abstract

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Background:

3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) is a key rate-limiting enzyme in the mevalonate pathway, which generates precursors for cholesterol biosynthesis and the production of non-steroidal mevalonate derivatives that are involved in a number of growth-regulatory processes. We have reported that lovastatin, a competitive inhibitor of HMG-CoA reductase, not only inhibits medulloblastoma proliferation in vitro, but also induces near-complete cell death via apoptosis. The present study explores some of the pathways which may be involved in lovastatin-induced apoptosis.

Methods:

Medulloblastoma cell lines were exposed in vitro to lovastatin with or without mevalonate, and document the effects using morphology, flow cytometry, DNA electrophoresis and Northern analysis.

Results:

1) Mevalonate prevents apoptosis when co-incubated with lovastatin, or when administered to lovastatin-pretreated cells. 2) Mevalonate restores the lovastatin-arrested cell cycle, allowing S phase entry. 3) Mevalonate does not prevent lovastatin-induced apoptosis after a critical duration of lovastatin pretreatment. For cell lines Daoy and UW228 this was 24 hours, and for D283 Med and D341 Med it was 48 hours. 4) Increases in HMG-CoA reductase mRNA levels induced by lovastatin are abrogated by co-incubation with lovastatin and mevalonate.

Conclusion:

These results confirm that lovastatin inhibition of this enzyme results in blockage of the mevalonate pathway, and that such a block is a critical step in the mechanism of lovastatin-induced apoptosis.

Résumé

RÉSUMÉIntroduction:

La 3-hydroxy-3-méthylglutaryl coenzyme a réductase (HMG-CoA réductase) estune enzyme clé, qui a un rôle d’étape cinétiquement limitante dans la voie du mévalonate et qui génère des précurseurs pour la biosynthèse du cholestérol et la production de dérivés non stéroïdiens du mévalonate impliqués dans certains processus de régulation de la croissance. Nous avons rapporté que la lovastatine, un inhibiteur compétitif de l’HMG-CoA réductase, non seulement inhibe la prolifération du médulloblastome in vitro, mais également induit presque complètement la mort cellulaire via l’apoptose. Cette étude explore certaines des voies qui pourraient être impliquées dans l’apoptose induite par la lovastatine.

Méthodes:

Les effets de l’exposition in vitro de lignées de cellules de médulloblastome à la lovastatine avec ou sans mévalonate ont été évalués par des etudes morphologiques, par cytométrie de flux, électrophorèse de l’ADN et analyse de Northern.

Résultats:

1) Le mévalonate prévient l’apoptose quand les cellules sont coincubées avec la lovastatine ou quand il est administré à des cellules prétraitées par la lovastatine. 2) Le mévalonate rétablit le cycle cellulaire interrompu par la lovastatine, permettant une entrée en phase S. 3) Le mévalonate ne prévient pas l’apoptose induite par la lovastatine après un temps de prétraitement critique par la lovastatine. Ce temps de prétraitement était de 24 heures pour les lignées cellulaires Daoy et UW228, et de 48 heures pour les lignées D283 Med et D341 Med. 4) L’augmentation des niveaux d’ARNm de l’HMG-CoA réductase induite par la lovastatine est abolie par la coincubation avec la lovastatine et le mévalonate.

Conclusions:

Ces résultats confirment que l’inhibition de cette enzyme par la lovastatine provoque un blocage de la voie métabolique du mévalonate et que ce blocage est une étape critique dans le mécanisme de l’apoptose induite par la lovastatine.

Type
Experimental Neurosciences
Information
Canadian Journal of Neurological Sciences , Volume 26 , Issue 4 , November 1999 , pp. 305 - 310
Copyright
Copyright © The Canadian Journal of Neurological 1999

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