TY - JOUR
T1 - Mechanism of action of α-naphthoflavone as an Ah receptor antagonist in MCF-7 human breast cancer cells
AU - Merchant, M.
AU - Krishnan, V.
AU - Safe, S.
PY - 1993/6
Y1 - 1993/6
N2 - α-Naphthoflavone (αNF) and 6-methyl-1,3,8-trichlorodibenzofuran (MCDF) inhibited 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced CYP1A1 gene expression in MCF-7 human breast cancer cells and also decreased the accumulation of the nuclear [3H]TCDD-aryl hydrocarbon (Ah) receptor complex. Nuclear extracts from cells treated with 10-6 M αNF and incubated with a dioxin responsive element (DRE, 26-mer) did not form a retarded band in a gel mobility shift assay. In contrast, incubation of nuclear extracts from cells treated with 10-6 M MCDF and DRE gave a retarded band and this is consistent with the antiestrogenic and Ah receptor agonist activity of MCDF in human breast cancer cells. αNF was further investigated as an Ah receptor antagonist by determining the inhibition by αNF of TCDD-induced antiestrogenicity in MCF-7 cells. TCDD (10-9 M) inhibited the 17β- estradiol-induced proliferation of MCF-7 cells and the secretion of the 52- kDa protein. In cotreatment studies, αNF (10-8 to 10-6 M) caused a concentration-dependent decrease in the antiestrogenic responses elicited by TCDD. In addition, αNF inhibited the TCDD-induced down-regulation of nuclear estrogen receptor levels in MCF-7 cells. αNF (10-6 M) alone was inactive as an estrogen or antiestrogen and in cotreatment studies did not affect 17β-estradiol-induced responses in MCF-7 cells. Tamoxifen (10-7 M), an antiestrogen which acts through the estrogen receptor, also inhibited 17β- estradiol-induced cell proliferation and αNF did not affect the tamoxifen- mediated antiproliferative response. Thus, αNF antagonized TCDD-induced CYP1A1 gene expression in MCF-7 cells and also acted as an anti-antiestrogen for TCDD-mediated antiestrogenicity in these cells. These results were consistent with the low levels of DRE binding observed with nuclear extracts from cells treated with 10-9 M TCDD plus αNF (10-8 to 10-6 M) or 10- 6 M αNF alone. Thus, αNF appears to act as an Ah receptor antagonist in MCF-7 cells by decreasing the levels of transcriptionally active nuclear Ah receptor complexes.
AB - α-Naphthoflavone (αNF) and 6-methyl-1,3,8-trichlorodibenzofuran (MCDF) inhibited 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced CYP1A1 gene expression in MCF-7 human breast cancer cells and also decreased the accumulation of the nuclear [3H]TCDD-aryl hydrocarbon (Ah) receptor complex. Nuclear extracts from cells treated with 10-6 M αNF and incubated with a dioxin responsive element (DRE, 26-mer) did not form a retarded band in a gel mobility shift assay. In contrast, incubation of nuclear extracts from cells treated with 10-6 M MCDF and DRE gave a retarded band and this is consistent with the antiestrogenic and Ah receptor agonist activity of MCDF in human breast cancer cells. αNF was further investigated as an Ah receptor antagonist by determining the inhibition by αNF of TCDD-induced antiestrogenicity in MCF-7 cells. TCDD (10-9 M) inhibited the 17β- estradiol-induced proliferation of MCF-7 cells and the secretion of the 52- kDa protein. In cotreatment studies, αNF (10-8 to 10-6 M) caused a concentration-dependent decrease in the antiestrogenic responses elicited by TCDD. In addition, αNF inhibited the TCDD-induced down-regulation of nuclear estrogen receptor levels in MCF-7 cells. αNF (10-6 M) alone was inactive as an estrogen or antiestrogen and in cotreatment studies did not affect 17β-estradiol-induced responses in MCF-7 cells. Tamoxifen (10-7 M), an antiestrogen which acts through the estrogen receptor, also inhibited 17β- estradiol-induced cell proliferation and αNF did not affect the tamoxifen- mediated antiproliferative response. Thus, αNF antagonized TCDD-induced CYP1A1 gene expression in MCF-7 cells and also acted as an anti-antiestrogen for TCDD-mediated antiestrogenicity in these cells. These results were consistent with the low levels of DRE binding observed with nuclear extracts from cells treated with 10-9 M TCDD plus αNF (10-8 to 10-6 M) or 10- 6 M αNF alone. Thus, αNF appears to act as an Ah receptor antagonist in MCF-7 cells by decreasing the levels of transcriptionally active nuclear Ah receptor complexes.
UR - http://www.scopus.com/inward/record.url?scp=0027235615&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027235615&partnerID=8YFLogxK
U2 - 10.1006/taap.1993.1101
DO - 10.1006/taap.1993.1101
M3 - Article
C2 - 8390116
AN - SCOPUS:0027235615
VL - 120
SP - 179
EP - 185
JO - Toxicology and Applied Pharmacology
JF - Toxicology and Applied Pharmacology
SN - 0041-008X
IS - 2
ER -