Effect of shRNA-mediated silence of H-ras gene on proliferation of human SACC-M cells / 中华口腔医学杂志
Chinese Journal of Stomatology
; (12): 113-117, 2008.
Article
em Zh
| WPRIM
| ID: wpr-235971
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To examine the effects of H-ras gene silence on cell cycle, proliferation and apoptosis of salivary adenoid cystic carcinoma -M (SACC-M) cell lines.</p><p><b>METHODS</b>The plasmid H-ras-shRNA, containing the sequence of shRNA targeting H-ras, and HK-shRNA (without interfering effect) were constructed and transfected into SACC-M cells. The cell line with shRNA plasmid stable expression was isolated by G418. The expression levels of H-ras were detected by RT-PCR and protein immunofluorescent assay; cell cycle and cell apoptosis were analyzed by flow cytometry (FCM). The proliferation of cell was also determined by subcutaneous tumor formation in nude mice.</p><p><b>RESULTS</b>After transfection of H-ras-shRNA plasmid, the mRNA expression of H-ras in SACC-M cells was down-regulated by 61.80% and protein expression of H-ras was inhibited by 62.76%; the cell proliferation was inhibited obviously; the G0G1 phase cells were increased. The cell apoptosis rate of H-ras-shRNA group was significantly higher than that of HK-shRNA group (P <0.05). The volume of subcutaneous tumor in nude mice was significantly smaller in Hras-shRNA group than in control group.</p><p><b>CONCLUSIONS</b>The recombinant plasmid HRAS-shRNA could efficiently down-regulate the expression of H-ras gene and protein, induce apoptosis of SACC-M cells and simultaneously inhibit proliferation of these cells in vitro and in vivo.</p>
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Base de dados:
WPRIM
Assunto principal:
Patologia
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Neoplasias das Glândulas Salivares
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Transfecção
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Proteínas Proto-Oncogênicas p21(ras)
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Apoptose
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Carcinoma Adenoide Cístico
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Inativação Gênica
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RNA Interferente Pequeno
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Linhagem Celular Tumoral
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Proliferação de Células
Limite:
Animals
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Female
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Humans
Idioma:
Zh
Ano de publicação:
2008
Tipo de documento:
Article