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

• Published in: Experimental & molecular medicine Vol. 40; no. 2; pp. 208 - 219
• Main Authors: Kang, Byung Young, Kim, Sujong, Lee, Ki-Hwan, Lee, Yong Sung, Hong, Il, Lee, Mi-Ock, Min, Daejin, Chang, Ihseop, Hwang, Jae Sung, Park, Jun Seong, Kim, Duck Hee, Kim, Byung-gee
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 30.04.2008; Springer Nature B.V; Korean Society of Medical Biochemistry and Molecular Biology
• Subjects: Base Sequence; Biomedical and Life Sciences; Biomedicine; Cell Line; DNA Primers; Enzyme-Linked Immunosorbent Assay; Gene Expression Profiling; Gene Expression Regulation - drug effects; Humans; Kaempferols - pharmacology; Keratinocytes - drug effects; Keratinocytes - metabolism; Medical Biochemistry; Molecular Medicine; Oligonucleotide Array Sequence Analysis; Original; Promoter Regions, Genetic; Reverse Transcriptase Polymerase Chain Reaction; Stem Cells; Transcription Factors - physiology; Transcription, Genetic - drug effects; NF-κB; keratinocytes; kaempferol; peroxisome proliferator-activated receptors; transcription factors; oligonucleotide array sequence analysis
• ISSN: 1226-3613; 2092-6413
• DOI: 10.3858/emm.2008.40.2.208

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-κB 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.