RNA-Binding Protein IGF2BP1 in Cutaneous Squamous Cell Carcinoma

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RNA-Binding Protein IGF2BP1 in Cutaneous Squamous Cell Carcinoma TaeWon Kim, Thomas Havighurst, KyungMann Kim, Scott J. Hebbring, Zhan Ye, Juliet Aylward, Sunduz Keles, Yaohui G. Xu, Vladimir S. Spiegelman Journal of Investigative Dermatology Volume 137, Issue 3, Pages 772-775 (March 2017) DOI: 10.1016/j.jid.2016.10.042 Copyright © 2016 The Authors Terms and Conditions

Figure 1 IGF2BP1 and its targets are overexpressed in SCC. (a) Boxplot of IGF2BP1 log2 fold change corresponding to Supplementary Figure S1 online. Mean fold change is 9.85 (23.30); median (dark line) is 11.00 (23.46) (P < 0.0001, one-sample Student t test). (b) Immunoblot of randomly selected cSCC samples from Supplementary Figure S1. (c–d) Scatterplots between expression level of IGF2BP1 mRNA and its targets, β-TrCP1 and c-Myc. The Spearman rank correlation coefficient and P-value are presented on each plot. A locally weighted scatterplot smoothing (LOWESS) smoothing curve shows the relationship between the variables. cSCC, cutaneous squamous cell carcinoma; Ctrl, control; SCC, squamous cell carcinoma. Journal of Investigative Dermatology 2017 137, 772-775DOI: (10.1016/j.jid.2016.10.042) Copyright © 2016 The Authors Terms and Conditions

Figure 2 Inhibition of IGF2BP1 suppressed SCC growth. (a) Ca3/7 (mouse) and SCC-13 (human SCC) were co-transfected either with scrambled sh-RNA (shScram.) and pTK-puro plasmid or with sh-IGF2BP1 and pTK-puro plasmid. Cells were treated with puromycin (Ca3/7 = 2.5 μg/ml; SCC-13 = 0.5 μg/ml) until colonies become visible to the naked eye. Colonies were stained with crystal violet. Mean ± standard deviation (n = 3, ∗P < 0.05, one-sample Student t test) are presented. (b) Ca3/7 and SCC-13 cells were co-transfected either with shScram. and Amaxa-GFP plasmid or with sh-IGF2BP1 and Amaxa-GFP plasmid. Annexin V-positive and GFP-positive cells were counted using flow cytometry. Mean ± standard deviation (n = 3, ∗P < 0.05, one-sample Student t test) are presented. (c) Immunoblot analysis of SCC cell lines transfected with IGF2BP1-specific shRNA. GFP, green fluorescent protein; SCC, squamous cell carcinoma; Scram, scrambled; sh, short hairpin. Journal of Investigative Dermatology 2017 137, 772-775DOI: (10.1016/j.jid.2016.10.042) Copyright © 2016 The Authors Terms and Conditions