Dose-response studies of the induction of hyperdiploidy and polyploidy by diethylstilbestrol and 17β-estradiol in cultured human lymphocytes using multicolor fluorescence in situ hybridization

Abstract

Diethylstilbestrol (DES) and 17β‐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 μM) and the steroid estrogen E2 (0.05–75 μM) 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 μM, whereas E2 showed its highest induction at 75 μM 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. Environ. Mol. Mutagen. 31:263–273, 1998 © 1998 Wiley‐Liss, Inc.

In Vitro Studies of Cellular and Molecular Developmental Toxicity of Adjuvants, Herbicides, and Fungicides Commonly Used in Red River Valley, Minnesota

“…The present study confirms these earlier findings and notes aneugenic effects as well. Aneuploidy and polyploidy reflect delayed or halted cell cycle progression, which may interrupt normal cell proliferation or trigger improper apoptosis in developing embryos, and is linked to stillbirths and birth defects (Oshimura & Barrett, 1986;Schuler et al, 1998). TPT was reported to induce aneuploidy in cultured human lymphocytes by altering spindle function (Jensen et al, 1991).…”

Nandrolone androgenic hormone presents genotoxic effects in different cells of mice

“…The testosterone derivatives could become aromatic and convert to 17 β ‐estradiol, which induces various chromosomal and genetic lesions, including aneuploidy, chromosomal aberrations, gene amplification, and microsatellite instability in cells in culture and/or in vivo and gene mutations in several cell test systems (Liehr, ). Some sex hormones such as β ‐estradiol, diethylstilbestrol and estriol (among others) have shown capacity to produce alterations in chromosome number (Wheeler et al , ; Schuler et al , ) and micronucleus induction by mitotic disturbances has been described (Eckert and Stopper, ; Fischer et al , ).…”

“…From the present study it is evident that the nandrolone induced genotoxic effects on peripheral blood, liver, bone marrow, brain and testicle cells of mice when tested using the comet assay, and induced clastogenic effects, as observed in the micronucleus test. Some studies showed that AAs do not induce gene mutation in classical bacterial and mammalian mutation assays, but at the chromosomal level several studies have reported that this similar hormones induces chromosomal breaks or aberrations (Schuler et al , ; Fischer et al , ; Torres‐Bugarín et al , ; among others).…”

“…Estrogens are also sex steroid hormones and, in general, do not induce gene mutation in classical bacterial and mammalian mutation assays (Roy and Liehr, ). However, at the chromosomal level of mutation, some studies have shown positive results (Banduhn and Obe, ; Wheeler et al , ; Eckert and Stopper, ; Schuler et al , ; Fischer et al , ; Torres‐Bugarín et al , ). Elevated levels of estrogens also are known to produce adverse effects, such as embryotoxicity, teratogenicity and carcinogenicity (IARC, ; Hertz, ; Marselos and Tomatis, ).…”

Detection of chromosomal aberrations by fluorescence in situ hybridization in cervicovaginal biopsies from women exposed to diethylstilbestrol in utero

“…Although estrogens have been clearly shown to be carcinogenic in human and animal models, the mechanism of action by which estrogenic compounds induce cancer is not well understood. Studies have demonstrated that DES induces morphologic and neoplastic transformation, as well as, aneuploidy in both in vivo and in vitro models (9,10,26,27) . Experimental evidence also suggests that DES is able to induce aneuploidy by binding to tubulin and microtubules and by subsequently interfering with spindle formation (28,29) .…”