Reversal of multi-drug resistance in K562/A02 cells by two short hairpin RNAs (shRNA) of mdr1 / 中华血液学杂志
Chinese Journal of Hematology
;
(12): 456-460, 2006.
Artigo
em Chinês
| WPRIM
| ID: wpr-243930
ABSTRACT
<p><b>OBJECTIVE</b>To construct two recombinant plasmids of mdr1 and mcl1 shRNA, and to investigate their reversal effect on drug resistance in K562 adriamycin resistant cell lines (K562/A02).</p><p><b>METHODS</b>Two oligonucleotides of mdr1 and mcl1 gene were designed referring to that of GenBank, double-stranded DNA was derived through annealing, and cloned into pRNAT vector digested by two restricted endoenzymes. K562/A02 cells were transfected with the recombinant plasmids. The mdr1 mRNA expression and its protein product P-glycoprotein (P-gp) were detected by RT-PCR and flow cytometry. The expression of mcl1 gene was detected by RT-PCR. 50% inhibition concentration (IC50) of adriamycin (ADM) on K562/A02 cells was determined by MTT method. Cells apoptosis was analyzed by flow cytometry.</p><p><b>RESULTS</b>Comparing with K562/A02 cells, the shRNA of mdrl or mcl1 gene in vitro can remarkably increase the sensitivity of K562/A02 to adriamycin, down-regulate mdr1 or mcl1 gene expression, increase the K562/A02 cells apoptosis rates induced by adriamycin. Cotransfection of mdrl and mcl1 genes shRNA can also down-regulate the expression of their gene, more remarkably increase the sensitivity and apoptosis of K562/ A02 to adriamycin.</p><p><b>CONCLUSION</b>Transfection of mdrl or mcl1 gene shRNA can promote the sensitivity of K562/A02 to adriamycin and cotransfection of the two shRNA can more remarkably do so. The mel1 gene might be involved in adriamycin resistant in K562/A02 cells.</p>
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Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
RNA Mensageiro
/
Transfecção
/
Expressão Gênica
/
Apoptose
/
Membro 1 da Subfamília B de Cassetes de Ligação de ATP
/
Resistência a Múltiplos Medicamentos
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Resistencia a Medicamentos Antineoplásicos
/
Proteínas Proto-Oncogênicas c-bcl-2
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Células K562
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Hematology
Ano de publicação:
2006
Tipo de documento:
Artigo
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