The effects of Hedgehog-Gli 1 signaling pathway on proliferation and apoptosis of hepatic stellate cells / 中华肝脏病杂志
Chinese Journal of Hepatology
; (12): 33-37, 2009.
Article
em Zh
| WPRIM
| ID: wpr-250063
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Hedgehog-Gli1 signaling pathway on proliferation, apoptosis and activation of hepatic stellate cells (HSCs) in vitro.</p><p><b>METHODS</b>The expression of Shh, Smo, Ptc and Gli-1 in HSC-T6 cells was analyzed by RT-PCR. HSC-T6 cells were incubated with various concentration of cyclopamine (0, 50, 100, 150, 200, 250 mumol/L) for 24 hours, cell viability was checked by MTT colorimetric assay, cell cycle was analyzed by flow cytometry, apoptosis was assayed by agarose electrophoresis of DNA and PI-Annexin V fluorescent staining, and the mRNA levels of Gli-1, TGF beta 1, PDGF and Bcl-2 were quantified by real-time RT-PCR.</p><p><b>RESULTS</b>RT-PCR indicated that the components of the Hedgehog-Gli1 signaling pathway were expressed in HSC-T6 cells. MTT assay indicated that cyclopamine inhibited cell viability in a concentration dependant manner (F = 636.81, P less than 0.01). Flow cytometry indicated that cells were piled up at G0/G1 phase in cyclopamine treated cells (65.08%+/-1.50%) as compared to control cells (55.41%+/-2.54%, t = -8.05, P less than 0.01). Cyclopamine treatment resulted in apoptosis as indicated by DNA fragmentation and PI-Annexin V staining. The mRNA levels of Gli-1, TGF beta 1, PDGF and Bcl-2 in cyclopamine treated cells were significantly lower than that in control cells (P less than 0.01).</p><p><b>CONCLUSION</b>Cyclopamine may inhibit the Hedgehog-Gli1 signaling, and hence repress proliferation and promote apoptosis in hepatic stellate cells.</p>
Texto completo:
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Base de dados:
WPRIM
Assunto principal:
Farmacologia
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Fatores de Transcrição
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Alcaloides de Veratrum
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Fator de Crescimento Derivado de Plaquetas
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RNA Mensageiro
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Transdução de Sinais
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Ciclo Celular
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Células Cultivadas
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Apoptose
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Reação em Cadeia da Polimerase Via Transcriptase Reversa
Limite:
Humans
Idioma:
Zh
Revista:
Chinese Journal of Hepatology
Ano de publicação:
2009
Tipo de documento:
Article