RESUMO
BACKGROUND: Recent studies suggest that androgens are the main regulator of changes in human hair growth, and dermal papilla (DP) is known to secret factors which regulate the growth and activity of the cells in the follicle in response to androgens. However, published data of androgen-regulated genes in human dermal papilla cells is limited and genome-wide large scale screening has not been reported. OBJECTIVE: To identify the dihydrotestosterone (DHT)-regulated genes in human dermal papilla cells. METHODS: SV40T-transformed human dermal papilla cell line (SV40T-DPC) was established and DHT-regulated genes were screened by cDNA microarray analysis. RESULTS: SV40T-DPC maintained early passage morphology and expressed functional androgen receptors. cDNA microarray followed by RT-PCR analysis showed that a number of genes are regulated by DHT in SV40T-DPC. CONCLUSION: The DHT-regulated genes reported in this study may be involved in androgen-mediated regulation of hair growth.
Assuntos
Antígenos Transformantes de Poliomavirus/metabolismo , Linhagem Celular Transformada , Derme/citologia , Di-Hidrotestosterona/metabolismo , Regulação da Expressão Gênica , Transporte Ativo do Núcleo Celular , Androgênios/metabolismo , DNA Complementar/metabolismo , Derme/metabolismo , Fibroblastos/metabolismo , Humanos , Imuno-Histoquímica/métodos , Microscopia de Fluorescência , Análise de Sequência com Séries de Oligonucleotídeos , Receptores Androgênicos/metabolismo , Pele/metabolismoRESUMO
To identify candidate genes that could be used as diagnostic and therapeutic targets for hepatocellular carcinoma (HCC), we searched for the genes that are overexpressed in HCC by combining representational difference analysis and microarray. Genes such as glypican-3 (GPC3), insulin-like growth factor 2, long-chain fatty-acid-coenzyme A ligase 4, farnesyl diphosphate synthase were frequently identified in our screening. Northern blot analysis with these four genes confirmed their overexpression in HCC. Among them we found that GPC3 transcript is upregulated in six out of seven cases of HCC. Immunoblot and immunohistochemical staining using polyclonal anti-GPC3 antibodies further confirmed that GPC3 protein is indeed increased in HCC tumor samples. We also found that GPC3 is secreted into culture media from cell lines derived from HCC. We conclude that GPC3 is a good molecular marker for HCC.
Assuntos
Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , Proteínas de Membrana/genética , Proteínas de Neoplasias/genética , Biomarcadores Tumorais/genética , Primers do DNA , Regulação Neoplásica da Expressão Gênica , Glipicanas , Humanos , Imuno-Histoquímica , Proteínas de Membrana/metabolismo , Proteínas de Neoplasias/metabolismo , Reação em Cadeia da Polimerase , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Tumorais CultivadasRESUMO
Urolithiasis and calcium oxalate crystal deposition diseases are still significant medical problems. In the course of nephrocalcin cDNA cloning, we have identified FKBP-12 as an inhibitory molecule of calcium oxalate crystal growth. lambdagt 11 cDNA libraries were constructed from renal carcinoma tissues and screened for nephrocalcin cDNA clones using anti-nephrocalcin antibody as a probe. Clones expressing recombinant proteins, which appeared to be antigenically cross-reactive to nephrocalcin, were isolated and their DNA sequences and inhibitory activities on the calcium oxalate crystal growth were determined. One of the clone lambda gt 11 #31-1 had a partial fragment (80 bp) of FKBP-12 cDNA as an insert. Therefore, a full-length FKBP-12 cDNA was PCR-cloned from the lambda gt 11 renal carcinoma cDNA library and was subcloned into an expression vector. The resultant recombinant FKBP-12 exhibited an inhibitory activity on the calcium oxalate crystal growth (Kd=10(-7) M). Physiological effect of the extracellular FKBP-12 was investigated in terms of macrophage activation and proinflammatory cytokine gene induction. Extracellular FKBP-12 failed to activate macrophages even at high concentrations. FKBP-12 seems an anti-stone molecule for the oxalate crystal deposition disease and recurrent stone diseases.