Estrogen-like activity of metals in MCF-7 breast cancer cells.
The ability of metals to activate estrogen receptor-alpha (ERalpha) was measured in the human breast cancer cell line, MCF-7. Similar to estradiol, treatment of cells with the divalent metals copper, cobalt, nickel, lead, mercury, tin, and chromium or with the metal anion vanadate stimulated cell proliferation; by d 6, there was a 2- to 5-fold increase in cell number. The metals also decreased the concentration of ERalpha protein and mRNA by 40-60% and induced expression of the estrogen-regulated genes progesterone receptor and pS2 by1.6- to 4-fold. Furthermore, there was a 2- to 4-fold increase in chloramphenicol acetyltransferase activity after treatment with the metals in COS-1 cells transiently cotransfected with the wild-type receptor and an estrogen-responsive chloramphenicol acetyltransferase reporter gene. The ability of the metals to alter gene expression was blocked by an antiestrogen, suggesting that the activity of these compounds is mediated by ERalpha. In binding assays the metals blocked the binding of estradiol to the receptor without altering the apparent binding affinity of the hormone (K(d) = 10(-10) M). Scatchard analysis employing either recombinant ERalpha or extracts from MCF-7 cells demonstrated that (57)Co and (63)Ni bind to ERalpha with equilibrium dissociation constants of 3 and 9.5 x 10(-9) and 2 and 7 x 10(-9) M, respectively. The ability of the metals to activate a chimeric receptor containing the hormone-binding domain of ERalpha suggests that their effects are mediated through the hormone-binding domain. Mutational analysis identified amino acids C381, C447, E523, H524, N532, and D538 as potential interaction sites, suggesting that divalent metals and metal anions activate ERalpha through the formation of a complex within the hormone-binding domain of the receptor.
Martin
Mary Beth
MB
Department of Biochemistry, Georgetown University, Washington, DC 20007, USA. martinmb@georgetown.edu
Reiter
Ronald
R
Pham
Trung
T
Avellanet
Yaniris R
YR
Camara
Johanna
J
Lahm
Michael
M
Pentecost
Elisabeth
E
Pratap
Kiran
K
Gilmore
Brent A
BA
Divekar
Shailaja
S
Dagata
Ross S
RS
Bull
Jaime L
JL
Stoica
Adriana
A
D000818:Animals; D001943:Breast Neoplasms; D019556:COS Cells; D002455:Cell Division / Q000187:drug effects; D002857:Chromium / Q000494:pharmacology; D003035:Cobalt / Q000494:pharmacology; D003300:Copper / Q000494:pharmacology; D047628:Estrogen Receptor alpha; D004967:Estrogens / Q000502:physiology*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007854:Lead / Q000494:pharmacology; D008628:Mercury / Q000494:pharmacology; D019216:Metals, Heavy / Q000494:pharmacology*; D009532:Nickel / Q000494:pharmacology; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism; D011980:Receptors, Progesterone / Q000235:genetics / Q000378:metabolism; D014001:Tin / Q000494:pharmacology; D000071163:Trefoil Factor-1; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D025521:Tumor Suppressor Proteins; D014638:Vanadates / Q000494:pharmacology
D047628:Estrogen Receptor alpha; D004967:Estrogens; D019216:Metals, Heavy; D011506:Proteins; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; C051838:TFF1 protein, human; D000071163:Trefoil Factor-1; D025521:Tumor Suppressor Proteins; D002857:Chromium; D007854:Lead; D003035:Cobalt; D014638:Vanadates; D014001:Tin; D003300:Copper; D009532:Nickel; D008628:Mercury