TY - JOUR
T1 - Genome-wide comparison of human keratinocyte and squamous cell carcinoma responses to UVB irradiation
T2 - Implications for skin and epithelial cancer
AU - Dazard, Jean Eudes
AU - Gal, Hilah
AU - Amariglio, Ninette
AU - Rechavi, Gideon
AU - Domany, Eytan
AU - Givol, David
N1 - Funding Information:
We are grateful to the family of Arison Dorsman for their donation to the Center for DNA Chips in the Pediatric Oncology Department, The Chaim Sheba Medical Center, Tel-Aviv. This study was supported in part by the Yad Abraham Research Center for Cancer Diagnosis and Therapy, and grants from the Irwin Green Alzheimer’s Research Fund, the Israel Science Foundation (ISF) and the Germany–Israel Science Foundation (GIF). We thank M Oren for monoclonal antibodies against p53 and MDM2, A Jetten for the polyclonal antibody against Cornifin, G de Murcia for the polyclonal antibody against PARP, and N Fusenig for HaCaT cells.
PY - 2003/5/15
Y1 - 2003/5/15
N2 - To gain insight into the transformation of epidermal cells into squamous carcinoma cells (SCC), we compared the response to ultraviolet B radiation (UVB) of normal human epidermal keratinocytes (NHEK) versus their transformed counterpart, SCC, using biological and molecular profiling. DNA microarray analyses (Affymetrix®, ∼ 12000 genes) indicated that the major group of upregulated genes in keratinocytes fall into three categories: (i) antiapoptotic and cell survival factors, including chemokines of the CXC/CC subfamilies (e.g. IL-8, GRO-1, -2, -3, SCYA20), growth factors (e.g. HB-EGF, CTGF, INSL-4), and proinflammatory mediators (e.g. COX-2, S100A9), (ii) DNA repair-related genes (e.g. GADD45, ERCC, BTG-1, Histories), and (iii) ECM proteases (MMP-1, -10). The major downregulated genes are ΔNp63 and PUMILIO, two potential markers for the maintenance of keratinocyte stem cells. NHEK were found to be more resistant than SCC to UVB-induced apoptosis and this resistance was mainly because of the protection from cell death by secreted survival factors, since it can be transferred from NHEK to SCC cultures by the conditioned medium. Whereas the response of keratinocytes to UVB involved regulation of key checkpoint genes (p53, MDM2, p21Cip1, ΔNp63), as well as antiapoptotic and DNA repair-related genes - no or little regulation of these genes was observed in SCC. The effect of UVB on NHEK and SCC resulted in upregulation of 251 and 127 genes, respectively, and downregulation of 322 genes in NHEK and 117 genes in SCC. To further analyse these changes, we used a novel unsupervised coupled two-way clustering method that allowed the identification of groups of genes that clearly partitioned keratinocytes from SCC, including a group of genes whose constitutive expression levels were similar before UVB. This allowed the identification of discriminating genes not otherwise revealed by simple static comparison in the absence of UVB irradiation. The implication of the changes in gene profile in keratinocytes for epithelial cancer is discussed.
AB - To gain insight into the transformation of epidermal cells into squamous carcinoma cells (SCC), we compared the response to ultraviolet B radiation (UVB) of normal human epidermal keratinocytes (NHEK) versus their transformed counterpart, SCC, using biological and molecular profiling. DNA microarray analyses (Affymetrix®, ∼ 12000 genes) indicated that the major group of upregulated genes in keratinocytes fall into three categories: (i) antiapoptotic and cell survival factors, including chemokines of the CXC/CC subfamilies (e.g. IL-8, GRO-1, -2, -3, SCYA20), growth factors (e.g. HB-EGF, CTGF, INSL-4), and proinflammatory mediators (e.g. COX-2, S100A9), (ii) DNA repair-related genes (e.g. GADD45, ERCC, BTG-1, Histories), and (iii) ECM proteases (MMP-1, -10). The major downregulated genes are ΔNp63 and PUMILIO, two potential markers for the maintenance of keratinocyte stem cells. NHEK were found to be more resistant than SCC to UVB-induced apoptosis and this resistance was mainly because of the protection from cell death by secreted survival factors, since it can be transferred from NHEK to SCC cultures by the conditioned medium. Whereas the response of keratinocytes to UVB involved regulation of key checkpoint genes (p53, MDM2, p21Cip1, ΔNp63), as well as antiapoptotic and DNA repair-related genes - no or little regulation of these genes was observed in SCC. The effect of UVB on NHEK and SCC resulted in upregulation of 251 and 127 genes, respectively, and downregulation of 322 genes in NHEK and 117 genes in SCC. To further analyse these changes, we used a novel unsupervised coupled two-way clustering method that allowed the identification of groups of genes that clearly partitioned keratinocytes from SCC, including a group of genes whose constitutive expression levels were similar before UVB. This allowed the identification of discriminating genes not otherwise revealed by simple static comparison in the absence of UVB irradiation. The implication of the changes in gene profile in keratinocytes for epithelial cancer is discussed.
KW - Apoptosis
KW - Chemokines
KW - Clustering
KW - Epidermis
KW - Microarray
UR - http://www.scopus.com/inward/record.url?scp=0037762589&partnerID=8YFLogxK
U2 - 10.1038/sj.onc.1206537
DO - 10.1038/sj.onc.1206537
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AN - SCOPUS:0037762589
SN - 0950-9232
VL - 22
SP - 2993
EP - 3006
JO - Oncogene
JF - Oncogene
IS - 19
ER -
