Pharmacotherapy Department, Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan; Global Career Design Center, Hiroshima University, Hiroshima, Japan; Pharmacology and Experimental Oncology Unit, Cancer Biology Department, National Cancer Institute, Cairo University, Cairo, Egypt.
Department of Pharmacology, Faculty of Veterinary Medicine, Kafrelsheikh University, Kafrelsheikh, Egypt; Department of Infectious Diseases, Graduate School of Medicine, International University of Health and Welfare, Narita, Japan.
Pharmacol Rep. 2019 Apr;71(2):289-298. doi: 10.1016/j.pharep.2019.01.001. Epub 2019 Jan 7.
Nor-wogonin, a polyhydroxy flavone, has been shown to possess antitumor activity. However, the mechanisms responsible for its antitumor activity are poorly studied. Herein, we investigated the mechanisms of nor-wogonin actions in triple-negative breast cancer (TNBC) cells.
Effects of nor-wogonin on cell proliferation and viability of four TNBC cell lines (MDA-MB-231, BT-549, HCC70, and HCC1806) and two non-tumorigenic breast cell lines (MCF-10A and AG11132) were assessed by BrdU incorporation assays and trypan blue dye exclusion tests. Cell cycle and apoptosis analyses were carried out by flow cytometry. Protein expression was analyzed by immunoblotting.
Nor-wogonin significantly inhibited the growth and decreased the viability of TNBC cells; however, it exhibited no or minimal effects in non-tumorigenic breast cells. Nor-wogonin (40 μM) was a more potent anti-proliferative and cytotoxic agent than wogonin (100 μM) and wogonoside (100 μM), which are structurally related to nor-wogonin. The antitumor effects of nor-wogonin can be attributed to cell cycle arrest via reduction of the expression of cyclin D1, cyclin B1, and CDK1. Furthermore, nor-wogonin induced mitochondrial apoptosis, (as evidenced by the increase in % of cells that are apoptotic), decreases in the mitochondrial membrane potential (ΔΨm), increases in Bax/Bcl-2 ratio, and caspase-3 cleavage. Moreover, nor-wogonin attenuated the expression of the nuclear factor kappa-B and activation of signal transducer and activator of transcription 3 pathways, which can be correlated with suppression of transforming growth factor-β-activated kinase 1 in TNBC cells.
These results showed that nor-wogonin might be a potential multi-target agent for TNBC treatment.
柚皮素是一种多羟基黄酮类化合物，已被证明具有抗肿瘤活性。然而，其抗肿瘤活性的机制尚未得到充分研究。在此，我们研究了柚皮素在三阴性乳腺癌（TNBC）细胞中的作用机制。
通过 BrdU 掺入试验和台盼蓝染料排除试验评估柚皮素对四种 TNBC 细胞系（MDA-MB-231、BT-549、HCC70 和 HCC1806）和两种非致瘤性乳腺细胞系（MCF-10A 和 AG11132）的增殖和活力的影响。通过流式细胞术进行细胞周期和凋亡分析。通过免疫印迹分析蛋白质表达。
柚皮素显著抑制 TNBC 细胞的生长并降低其活力，但对非致瘤性乳腺细胞几乎没有或没有影响。与结构上与柚皮素相关的柚皮苷（100 μM）和芹菜素苷（100 μM）相比，柚皮素（40 μM）是一种更有效的抗增殖和细胞毒性剂。柚皮素的抗肿瘤作用可归因于通过降低细胞周期蛋白 D1、细胞周期蛋白 B1 和 CDK1 的表达来抑制细胞周期。此外，柚皮素诱导线粒体凋亡（表现为凋亡细胞的百分比增加），降低线粒体膜电位（ΔΨm），增加 Bax/Bcl-2 比值和 caspase-3 切割。此外，柚皮素减弱了核因子 kappa-B 的表达和信号转导和转录激活因子 3 途径的激活，这与 TNBC 细胞中转化生长因子-β激活激酶 1 的抑制有关。
这些结果表明，柚皮素可能是 TNBC 治疗的潜在多靶标药物。
