Analysis of calcium-inducible genes in keratinocytes using suppression subtractive hybridization and cDNA microarray

Eun Young Seo, Jung Hyun Namkung, Kyu Mi Lee, Woong Hee Lee, Myung Im, Sun Ho Kee, Tae Park Geon, Jun Mo Yang, Young Joon Seo, Jang Kyu Park, Deok Kim Chang, Jeung Hoon Lee

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31 Citations (Scopus)

Abstract

Terminal differentiation of skin keratinocytes is a vertically directed multistep process that is tightly controlled by the sequential expression of a variety of genes. To gain further insight into the molecular events involved in this process, we used suppression subtraction hybridization (SSH) and cDNA microarray analysis. Messenger RNAs were isolated from primary skin keratinocytes cultured in vitro after treatment with calcium and then SSH was performed. A total of 840 cDNA clones were obtained from subtracted libraries, and these cDNA clones were used to make the microarray slides. Time-course cDNA microarray analysis (1, 3, 7, and 14 days after calcium treatment) revealed the global gene expression profile during keratinocyte differentiation. Of the 840 genes tested, 290 showed a greater than twofold change in expression level at least once over four time points. The genes were clustered into six groups according to their expression pattern using self-organizing map analysis and showed the global feature of function-related regulation. The genes related to keratinocyte differentiation were markedly up-regulated by calcium treatment. In addition, a unique pattern of increase was seen in the expression of genes related to ribosomal proteins. On the other hand, transcripts involved in metabolism, DNA repair, transcription, and translation were generally down-regulated. These results demonstrate the complexity of the gene expression profile that contributes to the spatiotemporal regulation of keratinocyte differentiation.

Original languageEnglish
Pages (from-to)528-538
Number of pages11
JournalGenomics
Volume86
Issue number5
DOIs
Publication statusPublished - 2005 Nov

Keywords

  • Calcium
  • Skin keratinocytes
  • Suppression subtraction hybridization
  • cDNA microarray

ASJC Scopus subject areas

  • Genetics

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    Seo, E. Y., Namkung, J. H., Lee, K. M., Lee, W. H., Im, M., Kee, S. H., Geon, T. P., Yang, J. M., Seo, Y. J., Park, J. K., Chang, D. K., & Lee, J. H. (2005). Analysis of calcium-inducible genes in keratinocytes using suppression subtractive hybridization and cDNA microarray. Genomics, 86(5), 528-538. https://doi.org/10.1016/j.ygeno.2005.06.013