Combined effect of xenoestrogens and growth factors in two estrogen-responsive cell lines.

It is now well recognized that estrogenic signaling mechanisms are far more complex than once thought. Several crosstalks between the estrogen receptor and other signaling pathways may influence the estrogenic stimulation of cell growth. Thus, the estrogenic effects of several environmental contaminants, now suspected to act as endocrine disrupters, may be influenced by a simultaneous stimulation of other signaling pathways. The aim of this study was to investigate whether the growth response of two estrogen-responsive cell lines, MCF-7 and GH3, treated with xenoestrogens might be affected by the addition of growth factors to their culture medium. Cells were treated with two known xenoestrogens, endosulfan and chlordane, alone or in the presence of insulin-like growth factor-1 and epidermal growth factor, respectively, and their growth was measured using the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxyanilide proliferation assay. Our results show that treatment with endosulfan or chlordane as well as treatment with growth factors increased cell growth, while the administration of xenoestrogens together with growth factors triggered a partly additive response with no antagonist or synergistic effect. These results sustain a role for xenoestrogens in cellular growth.

In vitro modulation of prolactin mRNA by toxaphene and 3,3',4,4'-tetrachlorobiphenyl.

It is now well recognized that many environmental contaminants are capable of disrupting endocrine processes in a variety of species, including birds, mammals, reptiles, and fish. Among these contaminants are toxaphene and polychlorinated biphenyls (PCBs), two of the most prevalent contaminants present in aquatic food chains of the Great Lakes and the Canadian Arctic region. We set out to investigate the possible endocrine-modulating activities of toxaphene, the PCB congener 3,3',4,4'-tetrachlorobiphenyl (TeCB), an equimolar mixture of both compounds (toxaphene/TeCB), and estradiol (E(2)) (E(2)/toxaphene, E(2)/TeCB) on prolactin (PRL) mRNA expression. Concentrations ranging from 10(-7) to 10(-11)M for both toxaphene and TeCB were assayed but only toxaphene modulated PRL mRNA levels, as determined by relative quantitative reverse transcriptase-polymerase chain reaction. Maximal induction by toxaphene was seen at 10(-7)M, resulting in a fourfold increase in PRL mRNA levels. No interactions were observed for combinations of the test substances. Our study demonstrates that toxaphene may exhibit estrogen-like activity by modulating PRL mRNA levels in GH(3) cells.

Modulation of prolactin expression by xenoestrogens.

Xenoestrogens are widely used environmental chemicals that have recently been under scrutiny because of their possible role as endocrine disrupters. Among them are endosulfan and chlordane, two persistent insecticides suspected to act as estrogens in living organisms. To test and better understand the potential estrogenic activity of these chemicals, we used a pituitary cell line (GH(3)) known to respond to estrogens by increasing its secretion of prolactin (PRL), a hormone that is well known for its many physiological functions, especially in fetal growth, development, and reproduction. We measured the levels of PRL secretion and PRL mRNA transcription using immunometric tests, Northern blots, and relative quantitative RT-PCR. We also employed the XTT proliferation assay to compare the growth of GH(3) cells stimulated with 17-beta estradiol and endosulfan or chlordane. Our results show that endosulfan and chlordane are able to induce a substantial increase of PRL expression while these two chemicals do not increase cell growth. Together, our results suggest that endosulfan and chlordane could indeed modulate an estrogen-inducible gene such as PRL, possibly acting via second messenger-mediated cellular mechanisms instead of solely competing with estrogens for the nuclear estrogen receptor sites.