Chen F-P, Chien M-H
Department of Obstetrics and Gynecology, Keelung Chang Gung Memorial Hospital, and Chang Gung University , Taiwan.
Climacteric. 2014 Aug;17(4):385-92. doi: 10.3109/13697137.2013.869671. Epub 2013 Dec 27.
To explore the effect and pathway of phytoestrogens on the growth of breast cancer cell line MCF-7.
MCF-7 cells (human estrogen receptor-positive and progesterone receptor-positive breast cancer cells) were cultured in serum-free medium for 24 h and then treated with genistein, resveratrol, and quercetin (10(-10)-10(-4) mol/l). After further incubation for 24, 48, 72, and 92 h, the cells were harvested and extracted for 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. According to the above results, the proteins involving proliferative and apoptotic pathways were evaluated by Western blot analysis.
Genistein, resveratrol, and quercetin significantly inhibited cellular proliferation in a dose- and time-dependent manner. Significantly elevated caspase-3 activity was noted after treatment with genistein (10(-9)-10(-7) mol/l), as well as with resveratrol and quercetin (10(-9)-10(-5) mol/l). Significant reduction of PI3K and AKT protein and significant increase of Fas ligand, Fas-associated protein with death domain, cytochrome C, truncated Bid, caspase-9, and caspase-3 were all noted after genistein, resveratrol, and quercetin treatment. 17β-Estradiol induced completely opposite results. Estrogen receptor (ER) α expression was significantly increased with 17β-estradiol, whereas ERβ expression was significantly elevated in the cultures with genistein, resveratrol, and quercetin.
These data demonstrate that genistein, resveratrol, and quercetin have antiproliferative effects on breast cancer cells. Their induction of apoptosis involves the activation of both the intrinsic and extrinsic apoptotic pathways, which may be related to the differential affinity to ERα and ERβ. Whether phytoestrogens have similar effects on normal breast cells remains to be examined.
探讨植物雌激素对乳腺癌细胞系MCF-7生长的影响及其作用途径。
将MCF-7细胞（人雌激素受体阳性和孕激素受体阳性乳腺癌细胞）在无血清培养基中培养24小时，然后用染料木黄酮、白藜芦醇和槲皮素（10⁻¹⁰ - 10⁻⁴mol/L）处理。进一步孵育24、48、72和92小时后，收获细胞并提取用于噻唑蓝（MTT）检测。根据上述结果，通过蛋白质免疫印迹分析评估参与增殖和凋亡途径的蛋白质。
染料木黄酮、白藜芦醇和槲皮素以剂量和时间依赖性方式显著抑制细胞增殖。在用染料木黄酮（10⁻⁹ - 10⁻⁷mol/L）以及白藜芦醇和槲皮素（10⁻⁹ - 10⁻⁵mol/L）处理后，观察到半胱天冬酶-3活性显著升高。在用染料木黄酮、白藜芦醇和槲皮素处理后，均观察到PI3K和AKT蛋白显著减少，而Fas配体、死亡结构域相关Fas蛋白、细胞色素C、截短的Bid、半胱天冬酶-9和半胱天冬酶-3显著增加。17β-雌二醇诱导出完全相反的结果。17β-雌二醇使雌激素受体（ER）α表达显著增加，而在含有染料木黄酮、白藜芦醇和槲皮素的培养物中ERβ表达显著升高。
这些数据表明，染料木黄酮、白藜芦醇和槲皮素对乳腺癌细胞具有抗增殖作用。它们诱导细胞凋亡涉及内源性和外源性凋亡途径的激活，这可能与对ERα和ERβ的不同亲和力有关。植物雌激素对正常乳腺细胞是否具有类似作用仍有待研究。
