Inhibition effect of antisense Bmi-1 on Jurkat cells / 中华血液学杂志
Chinese Journal of Hematology
;
(12): 554-556, 2005.
Artigo
em Chinês
| WPRIM
| ID: wpr-255841
ABSTRACT
<p><b>OBJECTIVES</b>To investigate whether antisense Bmi-1 plasmid could inhibit the proliferation of Jurkat cells.</p><p><b>METHODS</b>The antisense plasmid was constructed by PCR amplification of a 171 bp segment spanning Bmi-1 start codon and zinc finger structure and the PCR product was subsequently inserted reversely to plasmid pLNCX2. The final construct was confirmed through restriction enzyme digestion. G418 was added into the medium after the plasmid was successfully introduced into Jurkat cells by using lipofectin-mediated DNA transfection. The proliferation of Jurkat cells were determined by MTT and colony formation assays. Cell cycle was determined by flow cytometry. The p16 expression of Jurkat cells was studied by immunofluorescent histochemistry.</p><p><b>RESULTS</b>The growth rate of antisense Bmi-1 transfected Jurkat cells was significantly lower than that of the controls, and the colony forming capacity of the transfected cells decreased significantly (P < 0.01), the colony numbers being (90.7 +/- 9.07)/10(3) cells, (83.3 +/- 6.11)/10(3) cells and (56.0 +/- 5.56)/10(3) cells for control cells, empty plasmid transfected Jurkat cells and antisense Bmi-1 transfected Jurkat cells, respectively. The percentage of G, phase cells was increased and the p16 expression of antisense Bmi-1 transfected cells was significantly upregulated than that of control cells.</p><p><b>CONCLUSION</b>Antisense Bmi-1 can inhibit the growth and upregulate the expression of p16 of Jurkat cells in vitro.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Proteínas Repressoras
/
Proteínas Nucleares
/
Transfecção
/
Regulação Neoplásica da Expressão Gênica
/
Ciclo Celular
/
Oligonucleotídeos Antissenso
/
Proteínas Proto-Oncogênicas
/
Células Jurkat
/
Proliferação de Células
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Hematology
Ano de publicação:
2005
Tipo de documento:
Artigo
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