Chen F-P, Chien M-H
Department of Obstetrics and Gynecology, Keelung Chang Gung Memorial Hospital, and Chang Gung University , Taiwan.
Climacteric. 2014 Dec;17(6):682-91. doi: 10.3109/13697137.2014.937688. Epub 2014 Sep 19.
To explore the effect and pathway of phytoestrogens in vitro on the growth of both normal and malignant breast cells.
Normal breast MCF-10A cells and breast cancer MCF-7 cells were incubated with 10 (-10)-10(-4) mol/l genistein, resveratrol, and quercetin (plasma concentrations in human: 10 nmol/l-10 μmol/l) for 48 h and were then extracted for a cell proliferation assay (MTT), and for a cell death assay (TUNEL) assay. The proteins involved in the proliferative and apoptotic pathways were evaluated by Western blot analysis. Additionally, a comparison with 17β-estradiol as well as an evaluation of the differential effects on estrogen receptors (ER) α and β were performed.
MCF-7 cell proliferation was significantly inhibited at the concentrations greater than 10(-4) mol/l for all three phytoestrogens and from 10 (-5) mol/l for resveratrol and quercetin. MCF-10A cell proliferation was significantly increased at the concentrations from 10 (-8) to 10 (-5) mol/l for genistein and resveratrol and only at 10 (-5) mol/l for quercetin. Apoptotic cells were significantly increased by these phytoestrogens in the MCF-7 cells. At a concentration of 10 (-7) mol/l of these phytoestrogens, a significant reduction of PI3K and Akt and an increase of Fas ligand, Fas-associated protein with death domain, cytochrome C, truncated Bid, caspase-9, and caspase-3 were noted in the MCF-7; PI3K and Akt were significantly increased in the MCF-10A. ERβ expression was significantly elevated in MCF-10A and MCF-7 with these phytoestrogens. The effects of estradiol on normal and malignant breast cells were completely opposite to those of phytoestrogens.
This study demonstrates that phytoestrogens have antiproliferative effects on breast cancer cells via an ER-dependent mechanism, even at low concentrations, but are also capable of maintaining the survival of normal breast cell via ER-independent or other mechanisms.
探讨植物雌激素在体外对正常乳腺细胞和恶性乳腺细胞生长的影响及作用途径。
将正常乳腺MCF-10A细胞和乳腺癌MCF-7细胞分别用10（-10）-10（-4） mol/l的染料木黄酮、白藜芦醇和槲皮素（人体血浆浓度：10 nmol/l - 10 μmol/l）孵育48小时，然后提取细胞进行增殖测定（MTT法）和细胞死亡测定（TUNEL法）。通过蛋白质印迹分析评估参与增殖和凋亡途径的蛋白质。此外，与17β-雌二醇进行比较，并评估对雌激素受体（ER）α和β的不同影响。
对于所有三种植物雌激素，当浓度大于10（-4）mol/l时，MCF-7细胞增殖受到显著抑制；白藜芦醇和槲皮素在浓度为10（-5）mol/l时，MCF-7细胞增殖也受到显著抑制。对于染料木黄酮和白藜芦醇，当浓度在10（-8）至10（-5）mol/l时，MCF-10A细胞增殖显著增加；对于槲皮素，仅在10（-5）mol/l时，MCF-10A细胞增殖显著增加。这些植物雌激素使MCF-7细胞中的凋亡细胞显著增加。在这些植物雌激素浓度为10（-7） mol/l时，MCF-7细胞中PI3K和Akt显著减少，而Fas配体、死亡结构域相关蛋白Fas、细胞色素C、截短的Bid、半胱天冬酶-9和半胱天冬酶-3增加；在MCF-10A细胞中PI3K和Akt显著增加。这些植物雌激素使MCF-10A和MCF-7细胞中的ERβ表达显著升高。雌二醇对正常和恶性乳腺细胞的影响与植物雌激素完全相反。
本研究表明，植物雌激素即使在低浓度下也通过依赖雌激素受体的机制对乳腺癌细胞具有抗增殖作用，但也能够通过不依赖雌激素受体或其他机制维持正常乳腺细胞的存活。
