Table 2: Regulation of gene expression of tumor-related genes by epidermal growth factor (EGF).

NameControl
+EGF
Ratio-1+EGF
Control
Ratio-21/Ratio-2
Cy3Cy5Cy3Cy5

MMP12240104704.6726735790.224.61
Vimentin4596168773.67517018960.372.73
TIMP3275521130.7763114042.220.45
IGFBP36081.2013610.2240116434.090.24
BRAK717420890.2965446377.090.14

Total RNA was extracted with TRIzol reagent (Invitrogen) from cells treated with EGF for 24 h and untreated control cells and fluorescent cDNAs were prepared separately by a single round of reverse transcription in the presence of fluorescent Cy3-deoxy UTP for EGF-treated cells and Cy5-deoxy UTP for untreated ones. Cy3-deoxy UTP- and Cy5-deoxy UTP-labeled cDNAs were mixed and hybridized with IntelliGene Human Cytokine CHIP Ver. 3.0 (TAKARA). cDNA microarray analysis was performed according to the manufacturer’s protocol. The same experiment was repeated by reversing the labels, that is, using Cy5-deoxy UTP for EGF-treated cells and Cy3-deoxy UTP for the untreated ones. The hybridized chips were analyzed by ScanArray 4000 (Perkin-Elmer, Wellesley, MA, USA). MMP1: matrix metalloproteinase-1, TIMP3: tissue inhibitor of matrix metalloproteinase 3, IGFBP3: insulin-like growth factor-binding protein 3.