Data/hora: 11/06/2024 05:48:51

Provedor de dados: | 56
País: | Brazil
Título: | Secretory TAT-peptide-mediated protein transduction of LIF receptor α-chain distal cytoplasmic motifs into human myeloid HL-60 cells
Autores: | Sun,Q.; Xiong,J.; Lu,J.; Xu,S.; Li,Y.; Zhong,X.P.; Gao,G.K.; Liu,H.Q.
Data: | 2012-10-01
Ano: | 2012
Palavras-chave: | Leukemia inhibitory factor; TAT-HIV1; Protein transduction domain; Acute myeloid leukemia; LIF receptor.
Resumo: | The distal cytoplasmic motifs of leukemia inhibitory factor receptor α-chain (LIFRα-CT3) can independently induce intracellular myeloid differentiation in acute myeloid leukemia (AML) cells by gene transfection; however, there are significant limitations in the potential clinical use of these motifs due to liposome-derived genetic modifications. To produce a potentially therapeutic LIFRα-CT3 with cell-permeable activity, we constructed a eukaryotic expression pcDNA3.0-TAT-CT3-cMyc plasmid with a signal peptide (ss) inserted into the N-terminal that codes for an ss-TAT-CT3-cMyc fusion protein. The stable transfection of Chinese hamster ovary (CHO) cells via this vector and subsequent selection by Geneticin resulted in cell lines that express and secrete TAT-CT3-cMyc. The spent medium of pcDNA3.0-TAT-CT3-cMyc-transfected CHO cells could be purified using a cMyc-epitope-tag agarose affinity chromatography column and could be detected via SDS-PAGE, with antibodies against cMyc-tag. The direct administration of TAT-CT3-cMyc to HL-60 cell culture media caused the enrichment of CT3-cMyc in the cytoplasm and nucleus within 30 min and led to a significant reduction of viable cells (P &lt; 0.05) 8 h after exposure. The advantages of using this mammalian expression system include the ease of generating TAT fusion proteins that are adequately transcripted and the potential for a sustained production of such proteins in vitro for future AML therapy.
Tipo: | Info:eu-repo/semantics/article
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2012001000005
Editor: | Associação Brasileira de Divulgação Científica
Relação: | 10.1590/S0100-879X2012007500101
Formato: | text/html
Fonte: | Brazilian Journal of Medical and Biological Research  v.45 n.10 2012
Fonte: | info:eu-repo/semantics/openAccess