Antosiak Agata, Milowska Katarzyna, Maczynska Katarzyna, Rozalska Sylwia, Gabryelak Teresa
Department of General Biophysics, Faculty of Biology and Environmental Protection, University of Lodz, 141/143 Pomorska St., Lodz, 90-236 Poland.
Department of Industrial Microbiology and Biotechnology, University of Lodz, 12/16 Banacha St., Lodz, 90-237 Poland.
Med Chem Res. 2017;26(1):64-73. doi: 10.1007/s00044-016-1725-5. Epub 2016 Oct 3.
Genistein belongs to isoflavones, which are a subclass of flavonoids, a large group of polyphenolic compounds widely distributed in plants. Numerous in vitro studies suggest that isoflavones, particularly genistein, have both chemopreventive and chemotherapeutic potential in multiple tumor types. However, the molecular and cellular mechanisms of genistein effects on human ovarian cancer cells are still little known. In the present study, we investigated anticancer activity of genistein and its natural glucoside, genistein-8-C-glucoside isolated from flowers of L. We examined the effects of the two isoflavones alone or in combination on cultured human SK-OV-3 ovarian carcinoma cells. The cells were exposed to genistein and genistein-8-C-glucoside at various concentrations (1-90 µM) for 24 and 48 h. The cytotoxic and apoptotic properties of compounds were studied by the colorimetric 3-[4,5-2-yl]-2-5-diphenyltetrazolium bromide assay and the acridine orange/ethidium bromide staining technique. The morphological features of SK-OV-3 cells were examined by Nomarski differential interference contrast combined with a confocal laser scanning microscope. The level of ROS was evaluated with fluorescence probes: dichlorofluorescein-diacetate by flow cytometry. Changes in mitochondrial membrane potential were determined using 5,5,6,6-tetrachloro-1,1,3,3-tetraethylbenzimidazolcarbocyanine iodide. Genistein-treatment and genistein-8-C-glucoside-treatment resulted in the inhibition of cell proliferation, induction of apoptotic cell death and loss of mitochondrial membrane potential. The present data provide the first evidence in vitro that genistein-8-C-glucoside and combination genistein-genistein-8-C-glucoside could be a potential chemotherapeutic candidate for ovarian cancer therapy.
金雀异黄素属于异黄酮类，而异黄酮是黄酮类化合物的一个亚类，黄酮类是一大类广泛分布于植物中的多酚类化合物。大量体外研究表明，异黄酮，尤其是金雀异黄素，在多种肿瘤类型中具有化学预防和化疗潜力。然而，金雀异黄素对人卵巢癌细胞作用的分子和细胞机制仍鲜为人知。在本研究中，我们研究了金雀异黄素及其从L花中分离出的天然糖苷金雀异黄素-8-C-葡萄糖苷的抗癌活性。我们检测了这两种异黄酮单独或联合作用于培养的人SK-OV-3卵巢癌细胞的效果。将细胞暴露于不同浓度（1-90µM）的金雀异黄素和金雀异黄素-8-C-葡萄糖苷中24小时和48小时。通过比色法3-[4,5-2-基]-2-5-二苯基四氮唑溴盐法和吖啶橙/溴化乙锭染色技术研究化合物的细胞毒性和凋亡特性。通过诺马斯基微分干涉对比结合共聚焦激光扫描显微镜检查SK-OV-3细胞的形态特征。用荧光探针二氯荧光素二乙酸酯通过流式细胞术评估活性氧水平。使用5,5,6,6-四氯-1,1,3,3-四乙基苯并咪唑羰花青碘化物测定线粒体膜电位的变化。金雀异黄素处理和金雀异黄素-8-C-葡萄糖苷处理导致细胞增殖受到抑制、凋亡性细胞死亡诱导以及线粒体膜电位丧失。目前的数据首次在体外证明金雀异黄素-8-C-葡萄糖苷以及金雀异黄素与金雀异黄素-8-C-葡萄糖苷的组合可能是卵巢癌治疗的潜在化疗候选药物。
