Zoledronic acid activates the DNA S-phase checkpoint and induces osteosarcoma cell death characterized by apoptosis-inducing factor and endonuclease-G translocation independently of p53 and retinoblastoma status.
The molecular mechanisms responsible for the cellular effects of the nitrogen-containing bisphosphonate zoledronic acid (Zol) were assessed on several osteosarcoma cell lines differing in their p53 and retinoblastoma (Rb) status. Zol inhibited cell proliferation and increased atypical apoptosis. The Zol effects on proliferation were due to cell cycle arrest in S and G2/M phases subsequent to the activation of the intra-S DNA damage checkpoint with an increase in P-ATR, P-chk1, Wee1, and P-cdc2 levels and a decrease in cdc25c, regardless of the p53 and Rb status. In addition, the atypic apoptosis induced by Zol was independent of caspase activation, and it was characterized by nuclear alterations, increased Bax expression, and reduced Bcl-2 level. Furthermore, mitochondrial permeability was up-regulated by Zol independently of p53 in association with the translocation of apoptosis-inducing factor (AIF) and endonuclease-G (EndoG). Zol also disturbed cytoskeletal organization and cell junctions and inhibited cell migration and phosphorylation of focal adhesion kinases. The main difficulty encountered in treating cancer relates to mutations in key genes such as p53, Rb, or proteins affecting caspase signaling carried by many tumor cells. We have demonstrated for the first time that zoledronic acid activated the DNA damage S-phase checkpoint and the mitochondrial pathway via AIF and EndoG translocation, and it inhibited cell proliferation and induced cell death, bypassing these potentials mutations. Therefore, zoledronic acid may be considered as an effective therapeutic agent in clinical trials of osteosarcoma in which mutation for p53 and Rb very often occur, and where current treatment with traditional chemotherapeutic agents is ineffective.
Ory
B
B
Institut National de la Santé et de la Recherche Médicale ERI 7, 44035 Nantes cedex 1, France.
Blanchard
F
F
Battaglia
S
S
Gouin
F
F
Rédini
F
F
Heymann
D
D
D000818:Animals; D051033:Apoptosis Inducing Factor / Q000378:metabolism*; D050071:Bone Density Conservation Agents / Q000494:pharmacology; D001859:Bone Neoplasms / Q000473:pathology; D002453:Cell Cycle / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects*; D045744:Cell Line, Tumor; D004273:DNA, Neoplasm / Q000235:genetics*; D004164:Diphosphonates / Q000494:pharmacology*; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D018613:Microscopy, Confocal; D012516:Osteosarcoma / Q000473:pathology; D021381:Protein Transport; D051381:Rats; D016196:S Phase / Q000187:drug effects; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D000077211:Zoledronic Acid
D051033:Apoptosis Inducing Factor; D050071:Bone Density Conservation Agents; D004273:DNA, Neoplasm; D004164:Diphosphonates; D007093:Imidazoles; D016159:Tumor Suppressor Protein p53; D000077211:Zoledronic Acid; D004706:Endodeoxyribonucleases; C052391:endonuclease G