Cytotoxic properties of sunitinib and sorafenib on human corneal epithelial cells.
OBJECTIVE
To generate toxicology profiles of individual drugs on human corneal epithelial cells (HCEC) and compare their in vitro cytotoxicity.


METHODS
Monolayer cultures of HCEC were harvested from two human donor eyes. Sunitinib (0.3-10 µg/mL) and Sorafenib (0.3-100 µg/mL), diluted in culture medium (CnT-BM.1, CELLnTEC Advanced Cell Systems AG, Bern, Switzerland), 1% Penicillin and 1% Streptomycin were added to cells that were being grown in cell culture dishes. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was performed 24 hours, three days and five days after incubation. Live/dead viability/cytotoxicity assay (Live/dead assay) was performed and analyzed using fluorescence microscopy after 24 hours of incubation. The expression of p63, ABCG2 and PDGFRβ was evaluated by immunocytochemistry prior to exposure. Cell morphology was assessed with a phase contrast microscope after 24 hours of exposure.


RESULTS
Significant toxicity of Sunitinib was seen at concentrations of >3.3 µg/mL and of Sorafenib at concentrations of >1.0 µg/mL after 24 hours of incubation. Both drugs exhibited increasing toxicities over time. HCEC stained positively for p63, ABCG2 and PDGFRβ. In comparison, the IC50 (inhibitory concentration 50) of Sorafenib was 2.26 times the IC50 of Sunitinib using Live/dead assay after 24 hours and 2.39, 1.29 and 0.78 times the IC50 of Sunitinib using the MTT test after 24 hours, three days and five days, respectively.


CONCLUSIONS
These in vitro experimental findings support the safety of Sunitinib and Sorafenib on HCEC when used at a concentration of <3.3 µg/mL and <1.0 µg/mL, respectively, after 24 hours of exposure. The in vitro cytotoxicity of Sorafenib on HCEC was higher than Sunitinib.
Department of Ophthalmology, Eberhard-Karls University , Tuebingen , Germany.
Tarek
T
Bayyoud
Johanna
J
Hofmann
Martin
M
Spitzer
Karl Ulrich
KU
Bartz-Schmidt
Efdal
E
Yoeruek
D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000970:Antineoplastic Agents / Q000633:toxicity*; D015415:Biomarkers / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival; D004797:Enzyme-Linked Immunosorbent Assay; D019573:Epithelium, Corneal / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D007211:Indoles / Q000633:toxicity*; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D009363:Neoplasm Proteins / Q000378:metabolism; D009536:Niacinamide / Q000031:analogs & derivatives* / Q000633:toxicity; D010671:Phenylurea Compounds / Q000633:toxicity*; D047428:Protein Kinase Inhibitors / Q000633:toxicity*; D011758:Pyrroles / Q000633:toxicity*; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000378:metabolism; D000077157:Sorafenib; D000077210:Sunitinib; D013778:Tetrazolium Salts / Q000378:metabolism; D013844:Thiazoles / Q000378:metabolism
C116614:ABCG2 protein, human; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D000970:Antineoplastic Agents; D015415:Biomarkers; C081876:CKAP4 protein, human; D007211:Indoles; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D010671:Phenylurea Compounds; D047428:Protein Kinase Inhibitors; D011758:Pyrroles; D013778:Tetrazolium Salts; D013844:Thiazoles; D009536:Niacinamide; D000077157:Sorafenib; D020797:Receptor, Platelet-Derived Growth Factor beta; C022616:thiazolyl blue; D000077210:Sunitinib