Characterization of a point mutation in the hormone binding domain of the estrogen receptor from an breast tumor.
It is clear that growth of the MCF-7 breast cancer cell line is stimulated by estrogen and when estrogen is removed, growth slows. We have observed a tumor derived from MCF-7 cells that grows in athymic mice in the absence of estrogen stimulation. We hypothesized that a mutation in the estrogen receptor (ER) could be responsible for this constitutive growth. Using single stranded conformational polymorphism (SSCP) and DNA sequencing analysis, we have identified an ER containing a point mutation at position 415 (gly to val) within the hormone binding domain. The functional activity of this mutant was assessed in vitro and in vivo. Using transient transfection into an ER negative breast cancer cell with an ERE luciferase reporter gene, we found that both the wild-type and mutant receptors have similar efficacy. Additionally, the estrogenic responses were blocked by antiestrogens in a concentration related manner. We also found that tumors with the mutant receptor show similar growth response in athymic mice as wild-type: stimulation with estradiol and inhibition with antiestrogens. We conclude that the point mutation at position 415 (gly to val) is not responsible for constitutive growth.
England
G M
GM
Department of Surgery, Northwestern University, Chicago, IL 60611, USA.
Bilimoria
M M
MM
Chen
Z
Z
Assikis
V J
VJ
Muenzner
H D
HD
Jordan
V C
VC
D000818:Animals; D001665:Binding Sites; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology*; D002455:Cell Division / Q000187:drug effects; D017931:DNA Primers; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology*; D004965:Estrogen Antagonists / Q000494:pharmacology*; D005260:Female; D000077267:Fulvestrant; D005998:Glycine; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis; D051379:Mice; D008819:Mice, Nude; D010052:Ovariectomy; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D011960:Receptors, Estrogen / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects*; D014162:Transfection; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured; D014633:Valine
D017931:DNA Primers; D004965:Estrogen Antagonists; D011960:Receptors, Estrogen; D013629:Tamoxifen; C016601:afimoxifene; D000077267:Fulvestrant; D004958:Estradiol; D008156:Luciferases; D014633:Valine; D005998:Glycine