Effect of simvastatin on glioma cell proliferation, migration, and apoptosis.
OBJECTIVE
In this study, we investigated the effects of simvastatin on proliferation, migration, and apoptosis in human U251 and U87 glioma cells and the underlying molecular mechanism.


METHODS
We used colony formation assay to test the cell proliferation, in vitro scratch assay to examine the cell migration, and caspase-3 activity assay, annexin V staining, and cytochrome C release to evaluate the cell apoptosis. Lipid raft fractions were isolated from glioma cells. Total cholesterol content assay was used to test the change of cholesterol level in lipid raft fractions. Immunocytochemistry staining was performed to detect the changes of lipid rafts in cell membranes. Western blotting analysis was performed to examine the signal transduction both in cells and in lipid raft fractions.


RESULTS
Simvastatin inhibited proliferation and migration of U251 and U87 cells dose dependently. Simvastatin induced an increase of caspase-3 activity and annexin V staining, and down-regulated the phosphatidylinositol 3-kinase (PI3K)/Akt pathway. Simvastatin also decreased cholesterol content in lipid raft fractions, suppressed caveolin-1 expression in the lipid rafts, and induced Fas translocation into lipid rafts, suggesting that simvastatin may inhibit the prosurvival PI3K/Akt pathway and trigger caspase-3-dependent apoptotic cell death through the modulation of lipid rafts.


CONCLUSIONS
These results suggest that modulation of lipid rafts, Fas translocation, and PI3K/Akt/caspase-3 pathway are involved in the antitumor effect of simvastatin and may have a potential role in cancer prevention and treatment.
Wu
Hongtao
H
Department of Neurosurgery, Henry Ford Health System, Detroit, Michigan 48202, USA.
Jiang
Hao
H
Lu
Dunyue
D
Xiong
Ye
Y
Qu
Changsheng
C
Zhou
Dong
D
Mahmood
Asim
A
Chopp
Michael
M
D000704:Analysis of Variance; D017304:Annexin A5 / Q000378:metabolism; D000924:Anticholesteremic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D003114:Colony-Forming Units Assay / Q000379:methods; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005910:Glioma / Q000473:pathology* / Q000503:physiopathology; D006801:Humans; D021962:Membrane Microdomains / Q000378:metabolism / Q000473:pathology; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D019821:Simvastatin / Q000494:pharmacology*; D013997:Time Factors
D017304:Annexin A5; D000924:Anticholesteremic Agents; D004791:Enzyme Inhibitors; D019821:Simvastatin; D019869:Phosphatidylinositol 3-Kinases; D053148:Caspase 3