Common phytochemicals are ecdysteroid agonists and antagonists: a possible evolutionary link between vertebrate and invertebrate steroid hormones.

Many plant compounds are able to modulate growth and reproduction of herbivores by directly interacting with steroid hormone systems. In insects, several classes of phytochemicals, including the phytoestrogens, interfere with molting and reproduction. We investigated whether the anti-ecdysone activity may be due to interaction with the ecdysone receptor (EcR) using a reporter-gene assay and a cell differentiation assay of an ecdysone-responsive cell line, Cl.8+. We tested rutin (delays molt in insects); four flavones: luteolin and quercetin (metabolites of rutin), and apigenin and chrysin; and three non-flavones, coumestrol and genistein (both estrogenic) and tomatine (alters molt in insects). None of the phytochemicals tested were ecdysone agonists in the reporter-gene assay, but the flavones were able to significantly inhibit EcR-dependent gene transcription. In the Cl.8+ cells, quercetin and coumestrol were mixed agonists/antagonists, while genistein, tomatine and apigenin showed a synergistic effect with ecdysteroid in the reduction of cell growth. We suggest that the rutin effects on molting in insects are most likely due to the metabolites, luteolin or quercetin, while tomatine acts via a non-EcR pathway. Flavones not only interact with EcR and estrogen receptor (ER), but also signal nitrogen-fixing bacteria to form root nodules. The NodD protein which regulates this symbiosis has two ligand-binding domains similar to human ERalpha. The evolutionary significance of these findings are discussed.

Interaction of PAHs and PCBs with ecdysone-dependent gene expression and cell proliferation.

This study was done to determine whether PAHs and PCBs can interact with the arthropod steroid hormone system. Ecdysteroid molting hormones control growth, molting, and reproduction in arthropods. A spike in 20-OH ecdysone (20 HE) triggers the molt cycle in crustaceans, and earlier studies have shown that PAHs can affect this molt cycle in several crab species. However, the mechanism of this molt cycle interaction is unknown. Both PAHs and PCBs interact with other nuclear receptors; however, nothing is known about their ability to interact with the invertebrate ecdysone receptor (EcR). Four PAHs, benzo[a]pyrene, benzo[b]fluoranthene, pyrene, and chrysene, and the commercial PCB mixture, Aroclor 1254, were used to determine the ability of these classes of compounds to induce ecdysone-dependent reporter gene expression and to modify the proliferation and differentiation response of the ecdysteroid-responsive Cl.8+ cell line. The four PAHs were each able to enhance the ecdysteroid response in both the reporter gene and the cell proliferation assays only when given in conjunction with ecdysteroids. Aroclor 1254 had no effect in either system, either alone or in conjunction with ecdysteroids. These studies show that although the PAHs alone do not activate ecdysteroid-dependent gene expression or cell differentiation, they are able to enhance the effect of ecdysteroids, presumably through a non-receptor-mediated process. This mechanism may explain the effects on molting which have been reported after low-level crude oil exposures in crustaceans.