Construction of shRNA-Bmi-1 plasmid and its effect on proliferation of ECA109 cells / 中国医学科学院学报
Acta Academiae Medicinae Sinicae
;
(6): 17-22, 2015.
Artigo
em Inglês
| WPRIM
| ID: wpr-257687
ABSTRACT
<p><b>OBJECTIVE</b>To explore the role of Bmi-1 gene in the proliferation of squamous carcinoma cells and whether the silencing Bmi-1 can inhibit the growth of squamous cell carcinomas cells.</p><p><b>METHODS</b>The expressions of Bmi-1 in primary cultured Fibroblasts, karatinocyte cell line Hacat,squamous carcinoma cell line A431, and ECA109 were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. Recombinant plasmid inserted with Bmi-1 gene short hairpin RNA (shRNA) expression vector PGPU6/GFP/Neo-shBmi-1 was constructed and transfected into ECA109 cells with control set. After transfection for 48 and 72 hours,the mRNA and protein levels of Bmi-1 were examined with RT-PCR and Western blot analysis, respectively. The proliferation of the ECA109 cells was evaluated by MTT method and flow cytometry.</p><p><b>RESULTS</b>Bmi-1 was highly expressed in A431 and ECA109 cells than in Fibroblast cells and Hacat cells. The mRNA and protein expressions of Bmi-1 were significantly silenced in ECA109 cells after recombinant expression vector PGPU6/GFP/Neo-shBmi-1 transfection (P<0.05). Compared with the control groups,the proliferation of ECA109 transfected with PGPU6/GFP/Neo-shBmi-1 was significantly inhibited (P<0.05), and cells in G1 phase increased while in S phase decreased (P<0.05).</p><p><b>CONCLUSIONS</b>Bmi-1 is involved in the proliferation of squamous carcinoma cells. After the silencing of Bmi-1 expression,the proliferation ECA109 cells is suppressed due to the altered cell cycle.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
RNA Mensageiro
/
Carcinoma de Células Escamosas
/
Transfecção
/
Ciclo Celular
/
Linhagem Celular
/
Western Blotting
/
RNA Interferente Pequeno
/
Proliferação de Células
/
Fibroblastos
Limite:
Humanos
Idioma:
Inglês
Revista:
Acta Academiae Medicinae Sinicae
Ano de publicação:
2015
Tipo de documento:
Artigo
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