Institute of Biomedical Sciences Abel Salazar (ICBAS), Universidade do Porto (U.Porto), Rua de Jorge Viterbo Ferreira n 228, 4050-313 Porto, Portugal.
Interdisciplinary Center for Marine and Environmental Research (CIIMAR), Universidade do Porto (U.Porto), Avenida General Norton de Matos s/n, 4450-208 Matosinhos, Portugal.
Mar Drugs. 2019 Jul 30;17(8):448. doi: 10.3390/md17080448.
Preussin, a hydroxyl pyrrolidine derivative isolated from the marine sponge-associated fungus KUFA 0062, displayed anticancer effects in some cancer cell lines, including MCF7. Preussin was investigated for its cytotoxic and antiproliferative effects in breast cancer cell lines (MCF7, SKBR3, and MDA-MB-231), representatives of major breast cancers subtypes, and in a non-tumor cell line (MCF12A). Preussin was first tested in 2D (monolayer), and then in 3D (multicellular aggregates), cultures, using a multi-endpoint approach for cytotoxicity (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), resazurin and lactate dehydrogenase (LDH)) and proliferative (5-bromo-2'-deoxyuridine (BrdU)) assays, as well as the analysis of cell morphology by optical/electron microscopy and immunocytochemistry for caspase-3 and ki67. Preussin affected cell viability and proliferation in 2D and 3D cultures in all cell lines tested. The results in the 3D culture showed the same tendency as in the 2D culture, however, cells in the 3D culture were less responsive. The effects were observed at different concentrations of preussin, depending on the cell line and assay method. Morphological study of preussin-exposed cells revealed cell death, which was confirmed by caspase-3 immunostaining. In view of the data, we recommend a multi-endpoint approach, including histological evaluation, in future assays with the tested 3D models. Our data showed cytotoxic and antiproliferative activities of preussin in breast cancer cell lines in 2D and 3D cultures, warranting further studies for its anticancer potential.
普鲁辛，一种从海洋海绵相关真菌 KUFA 0062 中分离出来的羟基吡咯烷衍生物，在一些癌细胞系中表现出抗癌作用，包括 MCF7。普鲁辛在乳腺癌细胞系（MCF7、SKBR3 和 MDA-MB-231）和非肿瘤细胞系（MCF12A）中进行了细胞毒性和抗增殖作用的研究，这些细胞系代表了主要的乳腺癌亚型。普鲁辛首先在 2D（单层）培养物中进行了测试，然后在 3D（多细胞聚集）培养物中进行了测试，使用多终点方法进行细胞毒性（3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴盐（MTT）、resazurin 和乳酸脱氢酶（LDH））和增殖（5-溴-2'-脱氧尿苷（BrdU））测定，以及通过光学/电子显微镜和免疫细胞化学分析细胞形态来分析 caspase-3 和 ki67。普鲁辛在所有测试的细胞系的 2D 和 3D 培养物中均影响细胞活力和增殖。3D 培养物中的结果与 2D 培养物中的结果相同，但 3D 培养物中的细胞反应性较低。在不同浓度的普鲁辛作用下观察到了这些作用，这取决于细胞系和测定方法。暴露于普鲁辛的细胞的形态学研究显示细胞死亡，这通过 caspase-3 免疫染色得到了证实。鉴于这些数据，我们建议在未来的测试中采用多终点方法，包括组织学评估，以测试所使用的 3D 模型。我们的数据显示普鲁辛在 2D 和 3D 培养物中的乳腺癌细胞系中具有细胞毒性和抗增殖活性，值得进一步研究其抗癌潜力。
