Dose-response studies of the induction of hyperdiploidy and polyploidy by diethylstilbestrol and 17beta-estradiol in cultured human lymphocytes using multicolor fluorescence in situ hybridization.
Diethylstilbestrol (DES) and 17beta-estradiol (E2) are known inducers of aneuploidy and polyploidy in vivo and in vitro. Isolated human lymphocytes were treated with the stilbene estrogen DES (0.05-50 microM) and the steroid estrogen E2 (0.05-75 microM) in culture. Multicolor fluorescence in situ hybridization (FISH) with DNA probes for the centromere and adjacent heterochromatin regions of chromosomes 1, 9, and 16 was used to detect hyperdiploidy, polyploidy, and chromosomal breakage affecting these chromosomes. Using this FISH technique, significant nonlinear increases in hyperdiploidy were observed with both compounds, whereas no induction of chromosomal breakage affecting the pericentric heterochromatin regions of chromosomes 1, 9, and 16 could be detected. DES induced a maximum of approximately 13% hyperdiploid cells at 30 microM, whereas E2 showed its highest induction at 75 microM with 7% hyperdiploid cells. To distinguish hyperdiploidy from polyploidy, a FISH labeling strategy to detect multiple chromosomes simultaneously was established. Using this approach, we could show that most of the cells showing multiple hybridization regions after treatment with both chemicals were most likely the result of polyploidy rather than true hyperdiploidy. These results indicate that the induction of hyperdiploidy/polyploidy with DES and E2 show sublinear dose-response relationships with likely threshold concentrations in human lymphocytes and that FISH with multiple probes targeting different chromosomes can be used to estimate hyperdiploidy and polyploidy frequencies.
Schuler
M
M
Department of Entomology, University of California, Riverside 92521, USA.
Hasegawa
L
L
Parks
R
R
Metzler
M
M
Eastmond
D A
DA
D001562:Benzimidazoles; D001583:Benzoxazoles; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D002877:Chromosomes, Human / Q000187:drug effects* / Q000648:ultrastructure; D004054:Diethylstilbestrol / Q000008:administration & dosage / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000008:administration & dosage / Q000494:pharmacology*; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D008214:Lymphocytes / Q000187:drug effects* / Q000648:ultrastructure; D009942:Organometallic Compounds; D009943:Organophosphorus Compounds; D011123:Polyploidy; D011806:Quinolinium Compounds
D001562:Benzimidazoles; D001583:Benzoxazoles; D005456:Fluorescent Dyes; C104021:Fluoro-Green; D009942:Organometallic Compounds; D009943:Organophosphorus Compounds; D011806:Quinolinium Compounds; C066850:Cascade Blue; C075296:1,1'-((4,4,7,7-tetramethyl)-4,7-diazaundecamethylene)bis-4-(3-methyl-2,3-dihydro(benzo-1,3-oxazole)-2-methylidene)quinolinium; D004958:Estradiol; D004054:Diethylstilbestrol; D016650:Fluorescein-5-isothiocyanate