Exp Mol Med.  2008 Apr;40(2):208-219. 10.3858/emm.2008.40.2.208.

Transcriptional profiling in human HaCaT keratinocytes in response to kaempferol and identification of potential transcription factors for regulating differential gene expression

Affiliations
  • 1School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Korea. byungkim@snu.ac.kr
  • 2Skin Research Institute, AmorePacific Co. R&D Center, Kyounggi 449-729, Korea.
  • 3Department of Biochemistry, College of Medicine, Hanyang Universit, Seoul 133-791, Korea.
  • 4College of Pharmacy and Bio-MAX Institute, Seoul National University,Seoul 151-742, Korea.

Abstract

Kaempferol is the major flavonol in green tea and exhibits many biomedically useful properties such as antioxidative, cytoprotective and anti-apoptotic activities. To elucidate its effects on the skin, we investigated the transcriptional profiles of kaempferol-treated HaCaT cells using cDNA microarray analysis and identified 147 transcripts that exhibited significant changes in expression. Of these, 18 were up-regulated and 129 were down-regulated. These transcripts were then classified into 12 categories according to their functional roles: cell adhesion/cytoskeleton, cell cycle, redox homeostasis, immune/defense responses, metabolism, protein biosynthesis/modification, intracellular transport, RNA processing, DNA modification/ replication, regulation of transcription, signal transduction and transport. We then analyzed the promoter sequences of differentially-regulated genes and identified over-represented regulatory sites and candidate transcription factors (TFs) for gene regulation by kaempferol. These included c-REL, SAP-1, Ahr-ARNT, Nrf-2, Elk-1, SPI-B, NF-kappaB and p65. In addition, we validated the microarray results and promoter analyses using conventional methods such as real-time PCR and ELISA-based transcription factor assay. Our microarray analysis has provided useful information for determining the genetic regulatory network affected by kaempferol, and this approach will be useful for elucidating gene-phytochemical interactions.

Keyword

keratinocytes; kaempferol; NF-kappaB; oligonucleotide array sequence analysis; peroxisome proliferator-activated receptors; transcription factors
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