Establishment of screening system of environmental compounds for estrogenicity via WISP-2-induction

由于许多结构不同的化学物质被报道具有雌激素的功能，因此对化合物的雌激素活性的评估受到广泛关注。我们在人乳腺癌细胞株MCF-7中发现WISP-2(WNT-1可诱导信号通路蛋白2)是一个新的雌激素诱导基因。孕酮、地塞米松、三碘甲腺原氨酸和2，3，7，8-四氯二苯并对二恶英不能调节WISP-2的表达，表明其诱导是与雌激素受体相互作用的激素高度特异的。Western印迹分析显示，17α-雌二醇(17-雌二醇，E_2)诱导的WSAP-2蛋白不仅在细胞裂解产物中表达，而且在暴露细胞的培养上清液中也有表达，表明WIP-2是一种分泌型蛋白。E2对培养上清液中WSAP-2蛋白的诱导具有剂量依赖性，估计EC50和GT；水平在10到100 PM之间。这些结果证明了通过WISP-2诱导筛选环境化合物雌激素活性的能力。

As many structurally diverse chemicals have been reported to function as estrogens, evaluations for estrogenicity of compounds are of widespread concern. We identified WISP-2 (Wnt-1 inducible signaling pathway protein 2) as a novel estrogen-inducible gene in the MCF-7 human breast cancer cell lines. Progesterone, dexamethasone, tri-iodothyronine, and 2,3,7,8-tetrachlorodibenzo-p-dioxin did not regulate the expression of WISP-2, indicating that its induction is highly specific for hormones that interact with the estrogen receptor. Western blot analysis detected WISP-2 protein induced by 17α-estradiol(E2), not only in the cell lysates but also in the culture supernatant of exposed cells, indicating that WISP-2 was a secreted protein. The induction of WISP-2 protein by E2 in the culture supernatant was dose-dependent with estimated EC_<50> levels between 10 and 100 pM. These results demonstrated the capacity to screen environmental compounds for estrogenicity via WISP-2 induction.