Identification of calcium-inducible genes in primary keratinocytes using suppression-subtractive hybridization.
Terminal differentiation in epidermal keratinocytes involves major biochemical changes including the expression of many new differentiation-specific genes. To further understand this process, we performed suppression-subtractive hybridization of keratinocytes cultured under high-calcium condition, known to induce differentiation in vitro. We randomly isolated 300 clones representing 90 different genes. By reverse Northern blot analyses, 20 different genes were found to be overexpressed, of which 13 were confirmed as differentially expressed genes during keratinocyte differentiation by Northern blot analysis. Of those, five genes, transglutaminase 1, keratin 6, interleukin-1 receptor antagonist, kallikrein 7, and heat shock protein 27, are known to be up-regulated during epidermal differentiation. Six genes, ferritin-L chain, ribosomal protein S6, tumor-associated calcium signal transducer 2, neuroendocrine secretory protein 55, phosphoserine aminotransferase, and neutrophil gelatinase-associated lipocalin, heretofore were not known to be up-regulated during keratinocyte differentiation. We also identified two novel genes. One of these maps to chromosome 1q21 of the epidermal differentiation complex, and its expression level was strongly increased in differentiating keratinocytes. These differentially expressed genes may provide significant opportunities for further understanding of the epidermal keratinocyte differentiation.
Seo
Eun-Young
EY
Department of Dermatology, School of Medicine, Chungnam National University, Daejeon, South Korea.
Lee
Woong-Hee
WH
Piao
Yong-Jun
YJ
Kim
Kyoung-Hoon
KH
Lee
Kyu-Mi
KM
Ahn
Kwang-Sung
KS
Yang
Jun-Mo
JM
Seo
Young-Joon
YJ
Kim
Chang Deok
CD
Park
Jang-Kyu
JK
Lee
Jeung-Hoon
JH
D015152:Blotting, Northern; D002118:Calcium / Q000502:physiology*; D002478:Cells, Cultured; D000078404:Epidermal Cells; D004817:Epidermis / Q000502:physiology*; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D006824:Hybridization, Genetic; D015603:Keratinocytes / Q000166:cytology / Q000502:physiology*
D002118:Calcium