Stable expression of the human estrogen receptor in HeLa cells by infection: Effect of estrogen on cell proliferation and c- myc expression

Abstract

We have introduced the human estrogen receptor (ER) gene into HeLa cells, a human adenocarcinoma cell line of uterine origin, by infection. The ER cDNA was inserted into a retroviral vector (pMV7-ER) which also contains the neomycin resistance gene to allow for selection of stable infected clones. Northern analysis showed exogenous ER expression in stable clones. The ER protein expressed was about 66 kDa, similar to native MCF-7 ER, and binds with high affinity to estrogen (E 2). We have also observed that addition of E 2 at 10 −8 M inhibits the growth of the I-1 clone which expresses high levels of the ER (223 fmol/mg cytosol protein). The inhibitory effects of E 2 directly correlate with the quantity of ER in the cells. E 2-induced gene expression analysis showed that pS2 and progesterone receptor (PgR), genes induced in MCF-7 cells by E 2, are not induced in the ER + HeLa clones. However, c- myc expression was found to be decreased and may be responsible for the observed growth inhibition by E 2.