Construction of ERbeta expression vector and its function in different cancer cells / 中华肿瘤杂志
Chinese Journal of Oncology
;
(12): 340-343, 2003.
Artículo
en Chino
| WPRIM
| ID: wpr-347430
ABSTRACT
<p><b>OBJECTIVE</b>To construct an ERbeta expression vector and study its expression and function in different cancer cells.</p><p><b>METHODS</b>Standard PCR was used to amplify the full-length coding sequence of ERbeta. The amplified ERbeta gene was cloned into the eukaryotic expression vector pCDNA3, generating pCDNA3-ERbeta. The ERbeta expression was detected by Western blot and in vitro translation. The biological activity of ERbeta was detected by transfecting the pCDNA3-ERbeta into SV40-transformed embryonic kidney cell line 293T,breast cancer cell lines MDA-MB-435, MDA-MB-436, SKBR3, and prostate cancer cell line PC-3, with reporters containing estrogen response elements.</p><p><b>RESULTS</b>The recombinant plasmid pCDNA3-ERbeta was confirmed by restriction analysis to contain the ERbeta gene. The 63 000 ERbeta expression was shown by Western blot and further confirmed by in vitro translation. The ERbeta expression in different cancer cells was demonstrated to stimulate the expression of the reporters containing estrogen response elements, ERE and C3.</p><p><b>CONCLUSION</b>ERbeta protein is successfully expressed and has biological activity, laying solid foundation for further study on its role in cancer cells.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Patología
/
Plásmidos
/
Neoplasias de la Próstata
/
Proteínas Recombinantes
/
Neoplasias de la Mama
/
Transfección
/
Línea Celular
/
Genes Reporteros
/
Elementos de Respuesta
/
Biología Celular
Límite:
Femenino
/
Humanos
/
Masculino
Idioma:
Chino
Revista:
Chinese Journal of Oncology
Año:
2003
Tipo del documento:
Artículo
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