Cloning of expression vector of human tissue factor gene and its expression in human ovarian cancer cell line / 中国实验血液学杂志
Journal of Experimental Hematology
; (6): 579-582, 2003.
Article
en Zh
| WPRIM
| ID: wpr-278835
Biblioteca responsable:
WPRO
ABSTRACT
The aim was to construct the expressive vector of human tissue factor (TF), and determine its expressive level in stable-transfected human ovarian cancer cell line. The human TFcDNA was obtained from human placenta by RT-PCR and then inserted into eukaryotic expressive vector pcDNA3 to obtain the TF-pcDNA3 recombinant. This recombinant gene was introduced into human ovarian cell line A2780 through transfection mediated by lipofectamine. Stable-transfected cells were screened by G418. The TF expressive levels were detected by RT-PCR and flow cytometry. The results showed that: (1) the constructed product was identified as TF-pcDNA3 recombinant by sequencing. (2) TF was highly expressed not only at transcriptional level in the stable-transfected A2780 cell (transfected cell 3.99 +/- 0.15, untransfected cell 0.97 +/- 0.23, P < 0.01), but also on the membrane of the cell surface [transfected cell (48.56 +/- 9.53)%, untransfected cell (2.73 +/- 1.15)%, P < 0.01]. It was concluded that TF gene was successfully cloned, and was introduced into human ovarian cancer cell, and the subline A2780/TF which stably expresses TF at high level was obtained. It will provide good experimental basis for investigating new mechanisms of tumor growth, invasion, metastasis, hypercoagulability, and for exploring a new strategy of gene therapy.
Texto completo:
1
Índice:
WPRIM
Asunto principal:
Neoplasias Ováricas
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Patología
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Proteínas Recombinantes
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Tromboplastina
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Transfección
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Clonación Molecular
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Línea Celular Tumoral
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Genética
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Metabolismo
Tipo de estudio:
Prognostic_studies
Límite:
Animals
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Female
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Humans
Idioma:
Zh
Revista:
Journal of Experimental Hematology
Año:
2003
Tipo del documento:
Article