[Construction of ERbeta expression vector and its function in different cancer cells].
OBJECTIVE
To construct an ERbeta expression vector and study its expression and function in different cancer cells.


METHODS
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.


RESULTS
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.


CONCLUSIONS
ERbeta protein is successfully expressed and has biological activity, laying solid foundation for further study on its role in cancer cells.
Zhu
Jian-hua
JH
Beijing Institute of Biotechnology, Beijing 100850, China.
Ye
Qi-nong
QN
Jiang
Ze-fei
ZF
Zhong
Hong-jun
HJ
Yan
Jing-hua
JH
Lü
Qiu-jun
QJ
Song
San-tai
ST
Huang
Cui-fen
CF
D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D045744:Cell Line, Tumor; D004622:Embryo, Mammalian; D004847:Epithelial Cells; D047629:Estrogen Receptor beta / Q000235:genetics* / Q000378:metabolism*; D005260:Female; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D007668:Kidney / Q000166:cytology*; D008297:Male; D010957:Plasmids; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020218:Response Elements / Q000235:genetics; D014162:Transfection
D047629:Estrogen Receptor beta; D011994:Recombinant Proteins