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Bissan A. Ahmed, Andrew Stempel, Trevor McFarland, Bruce Ksander, Binoy Appukuttan, Tim Stout; Inhibition of Ocular Tumor Cell Growth With RTEF-1 Peptide Fragment. Invest. Ophthalmol. Vis. Sci. 2012;53(14):1222.
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Transcriptional enhancer factor 1-related (RTEF-1) is a member of the TEA DNA binding domain gene family. RTEF-1 is expressed within ocular vascular endothelial cells and has been shown to play a role in the control of VEGF expression. Ocular melanoma (OM) is the most common primary ocular tumor in adults , while Retinoblastoma (Rb) is a rapidly developing cancer of retina usually affecting children under the age of four. We previously identified a 651bp RTEF-1 isoform that can inhibit VEGF expression.The purpose of this study is to test whether a short peptide fragment derived from a functional domain of the 651bp RTEF-1 isoform can inhibit ocular tumor cell proliferation.
A 26 amino acid sequence corresponding to a Ser-Thr-Tyr domain within RTEF-1 linked to a 10 amino acid cell importation signal (RMR) was synthesized (GenScript U.S InC). Human ocular melanoma cells (Mel 202) (a kind gift from Dr Bryan Ksander) and Y79 retinoblasmotma cells (ATTC) were plated into a 96 well plate and cultured for 24 h. The STY-RMR peptide was added to the cell culture media at various concentrations (0.1 to 30 ug /100ul). Cell proliferation was assessed at 72 hours using a colorimetric XTT assay Cell proliferation was expressed as a percentage of the negative controls (n = 6). The amount of VEGF within media was determined by VEGF ELISA and compared between STY-RMR treated and controls (n = 6). Cellular apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) and by apoptosis ELISA .
Significant inhibition of ocular melanoma cell proliferation was obtained with 30ug/100ul of STY-RMR treatment (83%) (P < 0.01). A dose dependent response was also observed. Greater than 50% inhibition in cell proliferation was observed in the retinoblasmotma Y79 cells using the same dose (P < 0.01). A none significat increase in apoptosis of about (20%) could be detected in treated human ocular melanoma 24h after treatment.The optimum dose to obtain significant therapeutic effect varied depending on tumor type.
Using functional short peptide domains derived from the 651 RTEF-1 isoform may prove to be useful for treatment of ocular tumors.
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