Construction of the stable expression system of NIH3T3 fibroblast with pcDNA3.1(-)-hTGFbeta3 and the study on the proliferation of the system fibroblasts / 中华整形外科杂志
Chinese Journal of Plastic Surgery
;
(6): 109-112, 2006.
Artículo
en Chino
| WPRIM
| ID: wpr-240374
ABSTRACT
<p><b>OBJECTIVE</b>To construct and identify the stable expression system of NIH3T3 fibroblast with eukaryotic expression vector of human transforming growth factor beta3 (pcDNA3.1 (-)/TGFbeta3). So as to investigate the proliferation of NIH3T3 fibroblasts transfected with hTGFbeta3 gene stably.</p><p><b>METHODS</b>The stable transfection of NIH3T3 fibroblasts with recombinant plasmid expressing hTGFbeta3 was established by using LipofectamineTM2000 and G418 selection. The mRNA and protein expression of TGFbeta3 were detected by the RT-PCR and Western blot method, respectively. Microscope and MTT were adopted to examine the proliferation of the stable expression system of fibroblasts with hTGFbeta3.</p><p><b>RESULTS</b>After G418 selection, RT-PCR and Western blot analysis, 7 out of 10 cell lines transfected with pcDNA3.1 (-)/TGFbeta3 expressed with very high level of TGFbeta3, as compared with vector control transfectants that showed no expression, and compared with the other cell lines that expressed relatively low level. The stable transfection of NIH3T3 fibroblasts growth slowed down significantly (P < 0.05).</p><p><b>CONCLUSION</b>The stable expression system of NIH3T3 fibroblast with hTGFbeta3 were constructed successfully. The TGFbeta3 gene could inhibit the proliferation of NIH3T3 fibroblasts in vitro.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Plásmidos
/
Transfección
/
Células 3T3 NIH
/
Proliferación Celular
/
Factor de Crecimiento Transformador beta3
/
Fibroblastos
/
Genética
/
Metabolismo
Tipo de estudio:
Estudio pronóstico
Límite:
Animales
/
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Plastic Surgery
Año:
2006
Tipo del documento:
Artículo
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