Abstract | BACKGROUND: METHODS: Cell viability was tested by MTT assay. Autophagy was evaluated by western blot using anti-LC3 and anti-p62 antibodies, acridine orange staining and FACS analysis. Oxygen radical generation and necrosis were determined by FACS analysis using DCFDA and PI staining. Cathepsin B activation was evaluated by western blot and fluorogenic Z- Arg-Arg-AMC peptide. Finally, gene expression was performed using RT-PCR Profiler array. RESULTS: We found that sunitinib treatment for 24 h triggers incomplete autophagy, impairs cathepsin B activation and stimulates a lysosomal-dependent necrosis. By contrast, treatment for 48 h with pazopanib induces cathepsin B activation and autophagic cell death, markedly reversed by CA074-Me and 3-MA, cathepsin B and autophagic inhibitors, respectively. Finally, pazopanib upregulates the α- glucosidase and downregulates the TP73 mRNA expression. CONCLUSION: Our results showing distinct cell death mechanisms activated by different TKIs, provide the biological basis for novel molecularly targeted approaches.
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Authors | M Santoni, C Amantini, M B Morelli, S Liberati, V Farfariello, M Nabissi, L Bonfili, A M Eleuteri, M Mozzicafreddo, L Burattini, R Berardi, S Cascinu, G Santoni |
Journal | British journal of cancer
(Br J Cancer)
Vol. 109
Issue 4
Pg. 1040-50
(Aug 20 2013)
ISSN: 1532-1827 [Electronic] England |
PMID | 23887605
(Publication Type: Journal Article)
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Chemical References |
- Antineoplastic Agents
- Indazoles
- Indoles
- Pyrimidines
- Pyrroles
- Reactive Oxygen Species
- Sulfonamides
- pazopanib
- Protein-Tyrosine Kinases
- Sunitinib
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Topics |
- Antineoplastic Agents
(pharmacology)
- Autophagy
(drug effects)
- Carcinoma, Squamous Cell
(pathology)
- Carcinoma, Transitional Cell
(pathology)
- Cell Death
(drug effects)
- Cell Line, Tumor
- Cell Survival
(drug effects)
- Humans
- Indazoles
- Indoles
(pharmacology)
- Inhibitory Concentration 50
- Membrane Potential, Mitochondrial
(drug effects)
- Necrosis
(chemically induced)
- Protein-Tyrosine Kinases
(antagonists & inhibitors)
- Pyrimidines
(pharmacology)
- Pyrroles
(pharmacology)
- Reactive Oxygen Species
- Sulfonamides
(pharmacology)
- Sunitinib
- Urinary Bladder Neoplasms
(pathology)
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