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Assessment of estrogenic activity of natural compounds using improved E-screen assay
Chun-Yan Tian1, Chang-Qi Hu, Guang Xu
1Department of Molecular Genetics, School of Medicine, Fudan Univ ersity, Shanghai 200032, China. ytian@sina.com
Aim:
To improve E-screen assay and make it more accurate to screen estrogenic compounds.
Methods:
Estrogen receptor antisense RNA expression plasmid (pCASER) was constructed and introduced into MCF-7 with lipofectAMINE(TM), and positive clones were screened out with G418. PCR amplification was employed to identify whether estrogen receptor (ER) cDNA fragment had been inserted into MCF-7 cell genomes. Western blot was applied to detect the expression of ER. Cell growth was determined by MTT assay.
Results:
One ER antisense clone (MTASER) had been screened out. The effects of 17beta-estradiol, genistein, droloxifen, miyabenol C, and kobophenol A on MCF-7 were stronger than those effects on MTASER. Epidermal growth factor (EGF) had equivalent stimulatory effects on the proliferation of MCF-7 and MTASER.
Conclusion:
The improved E-screen assay could screen estrogenic compounds more accurately than original E-screen assay did.