Department of Biological Sciences, Florida Atlantic University, Davie, FL 33314, USA.
J Cell Mol Med. 2012 Nov;16(11):2631-6. doi: 10.1111/j.1582-4934.2012.01576.x.
Prostate cancer is one of the leading causes of death in men aged 40 to 55. Genistein isoflavone (4', 5', 7-trihydroxyisoflavone) is a dietary phytochemical with demonstrated anti-tumour activities in a variety of cancers. Topotecan Hydrochloride (Hycamtin) is an FDA-approved chemotherapy drug, primarily used for secondary treatment of ovarian, cervical and small cell lung cancers. This study was to demonstrate the potential anticancer efficacy of genistein-topotecan combination in LNCaP prostate cancer cells and the mechanism of the combination treatment. The LNCaP cells were grown in complete RPMI medium, and cultured at 37°C, 5% CO(2) for 24-48 hrs to achieve 70-90% confluency. The cells were treated with varying concentrations of genistein, topotecan and genistein-topotecan combination and incubated for 24 hrs. The treated cells were assayed for (i) post-treatment sensitivity using MTT assay and DNA fragmentation, (ii) treatment-induced apoptosis using caspase-3 and -9 binding assays and (iii) treatment-induced ROS generation levels. The overall data indicated that (i) both genistein and topotecan induce cellular death in LNCaP cells, (ii) genistein-topotecan combination was significantly more efficacious in reducing LNCaP cell viability compared with either genistein or topotecan alone, (iii) in all cases, cell death was primarily through apoptosis, via the activation of caspase-3 and -9, which are involved in the intrinsic pathway, (iv) ROS generation levels increased significantly with the genistein-topotecan combination treatment. Treatments involving genistein-topotecan combination may prove to be an attractive alternative phytotherapy or adjuvant therapy for prostate cancer.
前列腺癌是 40 至 55 岁男性死亡的主要原因之一。染料木黄酮（4'，5'，7-三羟基异黄酮）是一种膳食植物化学物质，已证明在多种癌症中有抗肿瘤活性。盐酸拓扑替康（Hycamtin）是一种 FDA 批准的化疗药物，主要用于卵巢癌、宫颈癌和小细胞肺癌的二线治疗。本研究旨在证明染料木黄酮-拓扑替康联合治疗在 LNCaP 前列腺癌细胞中的潜在抗癌疗效及其联合治疗的机制。LNCaP 细胞在完全 RPMI 培养基中生长，在 37°C、5%CO2 下培养 24-48 小时，达到 70-90%汇合度。用不同浓度的染料木黄酮、拓扑替康和染料木黄酮-拓扑替康联合处理细胞并孵育 24 小时。用 MTT 测定法和 DNA 片段化测定法检测处理后细胞的（i）敏感性，用 caspase-3 和 -9 结合测定法检测（ii）治疗诱导的细胞凋亡，用（iii）治疗诱导的 ROS 生成水平。总的数据表明：（i）染料木黄酮和拓扑替康都能诱导 LNCaP 细胞死亡，（ii）与单独使用染料木黄酮或拓扑替康相比，染料木黄酮-拓扑替康联合治疗显著更有效地降低 LNCaP 细胞活力，（iii）在所有情况下，细胞死亡主要通过凋亡，通过激活参与内在途径的 caspase-3 和 -9，（iv）ROS 生成水平随着染料木黄酮-拓扑替康联合治疗而显著增加。涉及染料木黄酮-拓扑替康联合治疗的方法可能被证明是一种有吸引力的替代植物疗法或辅助治疗前列腺癌的方法。
