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. 2017 Jan 23;6(1):e291. doi: 10.1038/oncsis.2016.88

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Copyright © 2017 The Author(s)

Oncogenesis is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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Integrative analysis of melanoma cells with overexpression and/or knock-down of CD271 links CD271 with DNA repair. (a) Venn diagram depicting commonly upregulated genes induced in A375 and T20/02 cells engineered to stably express CD271 (A375^NGFR/CD271, T20/02^NGFR/CD271) or stably transfected with a CD271-targeting shRNA (shCD271, T20/02^k.d.). Numbers of identified genes found commonly upregulated in A375^NGFR/CD271 and T20/02^NGFR/CD271 (235 genes) or differentially regulated in T20/02^NGFR/CD271 and T20/02^k.d. cells (577 genes, comprising up and downregulated genes) are shown. Right panel: DNA repair cluster from DAVID analysis of a set of 340 CD271-responsive genes upregulated in T20/02 cells, red or blue indicates positive or negative GO-contribution, respectively. (b) GSEA of profiling data of T20/02^NGFR/CD271 and T20/02^k.d. cells using a DNA repair signature (230 genes) as provided by MSigDB.¹⁸ (c) Verification of the CD271-dependent regulation of DNA repair genes RAD21, RAD51, RAD51AP1, DNA damage-binding protein 2 (DDB2) and xeroderma pigmentosum genes (XPC) in T20/02^NGFR/CD271 (left panel) or cells stably transfected with CD271 specific shRNAs (#2, #3, #4) by qPCR (right panel). Expression levels are shown in logarithmic scale, indicating relative expression levels±s.d. of independent triplicates, compared with cells either expressing GFP or knock-down control (shCtl.). *P⩽0.05; **P⩽0.01; ***P⩽0.001. (d) CD271-induced expression of DNA repair genes in A375^NGFR/CD271 cells as determined by qPCR. (e) Expression levels of CD271, RAD51AP1, RAD51 and MSH6 in cells of patient-tumor derived cell strains (Mel91, T20/02 and Mel41) sorted for presence or absence of CD271 by fluorescence-assisted cell sorting (FACS). Shown are three independent experiments.