Abstract
Osteosarcoma is the most common primary bone tumour in young adults. Despite improved prognosis, resistance to chemotherapy remains responsible for failure of osteosarcoma treatment. The identification of signals that promote apoptosis may provide clues to develop new therapeutic strategies for chemoresistant osteosarcoma. Here, we show that lipophilic statins (atorvastatin, simvastatin, cerivastatin) markedly induce caspases-dependent apoptosis in various human osteosarcoma cells, independently of bone morphogenetic protein (BMP)-2 signaling and cell differentiation. Although statins increased BMP-2 expression, the proapoptotic effect of statins was not prevented by the BMP antagonist noggin, and was abolished by mevalonate and geranylgeranylpyrophosphate, suggesting the involvement of defective protein geranylgeranylation. Consistently, lipophilic statins induced membrane RhoA relocalization to the cytosol and inhibited RhoA activity, which resulted in decreased phospho-p42/p44- mitogen-activated protein kinases (MAPKs) and Bcl-2 levels. Constitutively active RhoA rescued phospho-p42/p44-MAPKs and Bcl-2 and abolished statin-induced apoptosis. Thus, lipophilic statins induce caspase-dependent osteosarcoma cell apoptosis by a RhoA-p42/p44 MAPKs-Bcl-2-mediated mechanism, independently of BMP-2 signaling and cell differentiation.
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Abbreviations
- Ac-DEVD-pNA:
-
acetyl-Asp-Glu-Val-Asp-paranitroaniline
- BMP:
-
bone morphogenetic protein
- BMPR-IB:
-
BMP receptor IB
- BSA:
-
bovin serum albumin
- CA:
-
constitutively active form
- FCS:
-
fetal calf serum
- FPP:
-
farnesylpyrophosphate
- GAPDH:
-
glyceraldehyde 3-phosphate deshydrogenase
- GGPP:
-
geranylgeranylpyrophosphate
- HMG-CoA:
-
3-hydroxy 3-methylglutaryl coenzyme A
- MAPKs:
-
mitogen-activated protein kinases
- PI3K:
-
phosphoinositide 3′ kinase
- NO:
-
nitric oxide
- RT-PCR:
-
reverse transcription-polymerase chain reaction
- TBST:
-
tris hydroxymethylaminomethane buffered saline Tween-20
- WT:
-
wild-type form
- zVAD-fmk:
-
z-Val-Ala-Asp-fluoromethylketone
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Acknowledgements
This work was supported in part by Inserm, the Association pour la Recherche Contre le Cancer (ARC Contract #4315) and the Association Rhumatisme et Travail, Paris, France. We thank Drs. B Fournier and P Price (University of California, San Diego, La Jolla, CA, USA) for providing OHS4 cells, Dr. J Feyen (Bristol Squibb Meyer, USA) for providing statins, Dr. G Rawadi (Proskelia, Romainville, France) for the noggin vector, Drs. P Fort and P Jurdic (ENS, Lyon) for the RhoA constructs, Olivia Bawa for her contribution to this work and Nathalie Rochet (Inserm U364, Faculté de Médecine, Nice, France) for the gift of MG63, U2OS and CAL-72 osteosarcoma cells.
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Fromigué, O., Haÿ, E., Modrowski, D. et al. RhoA GTPase inactivation by statins induces osteosarcoma cell apoptosis by inhibiting p42/p44-MAPKs-Bcl-2 signaling independently of BMP-2 and cell differentiation. Cell Death Differ 13, 1845–1856 (2006). https://doi.org/10.1038/sj.cdd.4401873
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DOI: https://doi.org/10.1038/sj.cdd.4401873
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