Profiling of differentially expressed genes in human gingival epithelial cells and fibroblasts by DNA microarray.
Gingival epithelial cells and fibroblasts play important roles and have a harmonious relationship under normal and disease conditions, but the precise differences between theses cells remain unknown. To study the differences in gene expression between human gingival epithelial cells (HGE) and human gingival fibroblasts (HGF), mRNA was recovered from primary cultured cells and analyzed using cDNA microarray technology. The cDNA retro-transcribed from equal quantities of mRNA was labeled with the fluorescent dyes Cy5 and Cy3. The mixed probes were then hybridized with 7276 genes on the DNA microarray, after which fluorescence signals were scanned and further analyzed using GeneSpring software. Of the 7276 genes screened, 469 showed expression levels that were more than 2-fold greater in HGE than in HGF, while 293 showed expression levels that were more than 2-fold greater in HGF than in HGE. To confirm the reliability of the microarray results, keratin K5 and desmocolin, and vimentin and gp130, which showed higher mRNA levels in HGE and HGF, respectively, were selected and their mRNA levels were further analyzed by RT-PCR. The results of RT-PCR correlated well with those of microarray analysis. The present findings using a DNA microarray to detect differences in the gene expression profiles of HGE and HGF may be beneficial for genetic diagnosis of periodontal tissue metabolism and periodontal diseases.
Abiko
Yoshimitsu
Y
Department of Biochemistry, Nihon University School of Dentistry at Matsudo, Chiba, Japan. yabiko@mascat.nihon-u.ac.jp
Hiratsuka
Koichi
K
Kiyama-Kishikawa
Michiko
M
Tsushima
Katsumasa
K
Ohta
Mitsuhiro
M
Sasahara
Hiroshige
H
D015703:Antigens, CD / Q000235:genetics; D002478:Cells, Cultured; D050822:Cytokine Receptor gp130; D051187:Desmocollins; D003896:Desmosomes / Q000235:genetics; D004847:Epithelial Cells / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D005456:Fluorescent Dyes; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000235:genetics*; D005881:Gingiva / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D053555:Keratin-5; D007633:Keratins / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics; D018121:Receptors, Cytokine / Q000235:genetics; D053677:Receptors, Oncostatin M; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics; D014746:Vimentin / Q000235:genetics
D015703:Antigens, CD; C081561:DSC2 protein, human; D051187:Desmocollins; D005456:Fluorescent Dyes; C494199:IL6ST protein, human; C503940:KRT5 protein, human; D053555:Keratin-5; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D018121:Receptors, Cytokine; D053677:Receptors, Oncostatin M; D014746:Vimentin; D050822:Cytokine Receptor gp130; D007633:Keratins