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a CNRS UMR 8161, Institut de Biologie de Lille, CNRS/Institut Pasteur de Lille/Université de Lille I/Université de Lille II, B.P. 447, 59021 Lille Cedex, France b Plate-forme Biopuces, Université de Lille 2, Faculté de Médecine, Place de Verdun, 59045 Lille, France
Key Words: HGF/SF • MET • apoptosis • target genes • breast • array
Address for correspondence: Véronique Fafeur, CNRS UMR 8161 Institut de Biologie de Lille, Institut Pasteur de Lille, B.P. 447, 59021 Lille Cedex, France. Voice: 33-3-20-87-10-91; fax: 33-3-20-87-11-11. e-mail: veronique.fafeur{at}ibl.fr
Hepatocyte growth factor/scatter factor (HGF/SF) induces scattering, morphogenesis, and survival of epithelial cells through activation of the MET tyrosine kinase receptor. HGF/SF and MET are involved in normal development and tumor progression of many tissues and organs, including the mammary gland. In order to find target genes of HGF/SF involved in its survival function, we used an oligonucleotide microarray representing 1,920 genes known to be involved in apoptosis, transcriptional regulation, and signal transduction. MCF-10A human mammary epithelial cells were grown in the absence of serum and treated or not with HGF/SF for 2 h. Total RNA was reverse-transcribed to cDNA in the presence of fluorescent Cy3-dUTP or Cy5-dUTP to generate fluorescently labeled cDNA probes. Microarrays were performed and the ratios of Cy5/Cy3 fluorescence were determined. The expression of three apoptotic genes was modified by HGF/SF, with A20 being upregulated, and DAXX and SMAC being downregulated. These changes of expression were confirmed by real-time quantitative PCR. According to current-knowledge, A20 is antiapoptotic and SMAC is proapoptotic, while a pro- or antiapoptotic function of DAXX is controversial. The fact that HGF/SF upregulates an antiapoptotic gene (A20) and downregulates a proapoptotic gene (SMAC) is in agreement with its survival effect in MCF-10A cells. This study identified novel apoptotic genes regulated by HGF/SF, which can contribute to its survival effect.
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