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Cell-Cycle Gene Expression in Lovastatin-Induced Medulloblastoma Apoptosis

Published online by Cambridge University Press:  02 December 2014

Wei Wang  and
Robert J.B. Macaulay
Wei Wang
Affiliation:
Laboratory of Cellular Oncology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA
Robert J.B. Macaulay
Affiliation:
Department of Pathology and Laboratory Medicine, QEII Health Sciences Center and Dalhousie University, Halifax, Nova Scotia B3H 1V8, Canada
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Abstract

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Background:

3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase is a key rate-limiting enzyme in the mevalonate pathway, which generates precursors both for cholesterol biosynthesis and for the production of nonsteroidal 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 mechanism of this phenomenon is unclear. Possible involvement of changes in expression of certain cell-cycle related genes led us to study some of them in more detail.

Methods:

Medulloblastoma cell lines were exposed in vitro to lovastatin, and the effects of gene expression changes were studied using RT-PCR, antisense oligonucleotide, DNA electrophoresis and Western blotting analysis.

Results:

1) Levels of total Ras gene mRNA and individual Ras gene mRNA are stable in lovastatin treatment in all examined medulloblastoma cell lines. 2) Blocking c-myc gene over-expression does not enhance medulloblastoma cell sensitivity to lovastatin. 3) Following lovastatin treatment, p16 expression exhibits no change, but pronounced increases of p27KIP1 protein are observed in all examined cell lines. Lovastatin induces pronounced increases of p21WAF1 protein only in Daoy and UW228, but not in D283 Med and D341 Med. 4) Following lovastatin treatment, increased p53 protein is detected only in D341 Med, and bax protein is unchanged in all cell lines.

Conclusion:

Lovastatin-induced growth inhibition and apoptosis in medulloblastoma are not dependent on the regulation of Ras and c-myc gene expression, but may be mediated by p27KIP1 gene expression. Lovastatin-induced apoptosis in medulloblastoma is probably p53 independent, but p53 and p21WAF1 gene expression may also mediate anti-proliferative effects of lovastatin on specific medulloblastoma cell lines.

Résumé:

RÉSUMÉ:Introduction:

La 3-hydroxy-3-méthylglutaryl-coenzyme A (HGM-CoA) réductase est un enzyme limitant clé de la voie du mévalonate qui génère des précurseurs tant pour la biosynthèse du cholestérol que pour la production de dérivés non stéroïdiens du mévalonate qui sont impliqués dans certains processus régulateurs de la croissance. Nous avons rapporté que la lovastatine, un inhibiteur non compétitif de l'HMG-CoA réductase, inhibe la prolifération du médulloblastome in vitro et induit également la mort cellulaire presque complète via l'apoptose. Le mécanisme sous-jacent à ce phénomène n'est pas clair. La possibilité que des changements dans l'expression de certains gènes du cycle cellulaire soient impliqués nous a incités à en étudier quelques-uns de plus près.

Méthodes:

Des lignées cellulaires de médulloblastome ont été exposées in vitro à la lovastatine et les effets des changements dans l'expression génique ont été étudiés au moyen de RT-PCR, d'oligonucléotides antisenses, de l'électrophorèse de l'ADN et du buvardage western.

Résultats:

Les niveaux d'ARNm de tous les gènes Ras et des gènes Ras pris individuellement sont stables dans toutes les lignées cellulaires de médulloblastome étudiées après traitement par la lovastatine. 2) Le fait de bloquer la surexpression du gène c-myc n'augmente pas la sensibilité des cellules de médulloblastome à la lovastatine. 3) Suite au traitement par la lovastatine, l'expression de p16 ne change pas, mais on observe des augmentations considérables de la protéine p27KIP1 dans toutes les lignées cellulaires examinées. La lovastatine provoque des augmentations marquées de la protéine p21WAF1 dans le lignées Daoy et UW228, mais pas dans D283 Med et D341 Med. 4) Suite au traitement par la lovastatine, une augmentation de la protéine p53 est détectée seulement dans D341 Med et la protéine Box demeure inchangée dans toutes les lignées cellulaires.

Conclusions:

L'inhibition de la croissance et l'apoptose induites par la lovastatine dans le médulloblastome ne sont pas dépendantes de la régulation de l'expression des gènes Ras et c-myc, mais elles pourraient être médiées par l'expression du gène p27KIP1. L'apoptose induite par la lovastatine dans le médulloblastome est probablement indépendante de p53, mais l'expression des gènes p53 et p21WAF1 peut également médier des effets antiprolifératifs de la lovastatine dans des lignées cellulaires spécifiques de médulloblastome.

Type
Research Article
Information
Canadian Journal of Neurological Sciences , Volume 30 , Issue 4 , November 2003 , pp. 349 - 357
Copyright
Copyright © The Canadian Journal of Neurological 2003

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