Original Article

Assessment of estrogenic activity of natural compounds using improved E-screen assay.

Authors: Chun-Yan TIAN, Chang-Qi HU, Guang XU, Hou-Yan SONG

Abstract

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.
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