Hastings Jordan, Kenealey Jason
Department of Nutrition, Dietetics, and Food Science, Brigham Young University, ESC S249, Provo, UT 84602 USA.
Cancer Cell Int. 2017 Oct 18;17:93. doi: 10.1186/s12935-017-0464-0. eCollection 2017.
Avenanthramides (AVN) are a relatively unstudied family of phytochemicals that could be novel chemotherapeutics. These compounds, found in oats, are non-toxic to healthy cells and have been shown to reduce viability of human colon and liver cancers in vitro. However, these studies do not elucidate a molecular mechanism for individual AVN. In this study we aim to see the effects of AVN on MDA-MB-231 breast cancer cells.
An MTT assay was used to determine cell viability. Staining and analysis with a flow cytometer was used to identify cell cycle progression and apoptosis. FloJo software was used to analyze the cytometric data. In all experiments, statistical significance was determined by a two-tailed t test.
This study demonstrates that AVN-A, B, and C individually reduce viability in the MDA-MB-231 breast cancer cell line. AVN-C has the most potent decrease in tumor cell viability, decreasing viable cells to below 25% at 400 µM when compared to control after 96 h. We demonstrate that treatment with AVN-C causes DNA fragmentation and accumulation of over 90% of cells into a sub G cell cycle population. Further, we conclude that AVN-C treated cells activate apoptosis because 97% of treated cells stain positive for annexin V while 91% have caspase-3/7 activity, a late marker of apoptosis.
Breast cancer cells treated with AVN-C have a decrease in cell viability, an increase in the sub G population, and stain positive for both annexin V and caspase activity, indicating that AVN-C induces apoptosis in breast cancer cells. These compounds may be able to act as chemotherapeutics as demonstrated through future in vivo studies.
燕麦酰胺(AVN)是一类研究相对较少的植物化学物质,可能是新型的化学治疗剂。这些存在于燕麦中的化合物对健康细胞无毒,并且已显示在体外可降低人结肠癌和肝癌细胞的活力。然而,这些研究并未阐明单个燕麦酰胺的分子机制。在本研究中,我们旨在观察燕麦酰胺对MDA-MB-231乳腺癌细胞的影响。
采用MTT法测定细胞活力。使用流式细胞仪进行染色和分析以确定细胞周期进程和细胞凋亡。使用FloJo软件分析流式细胞术数据。在所有实验中,通过双侧t检验确定统计学显著性。
本研究表明,燕麦酰胺A、B和C分别降低MDA-MB-231乳腺癌细胞系的活力。燕麦酰胺C对肿瘤细胞活力的降低作用最强,在96小时后,与对照组相比,400µM时可将活细胞减少至25%以下。我们证明,用燕麦酰胺C处理会导致DNA片段化,并使超过90%的细胞积累到亚G期细胞周期群体中。此外,我们得出结论,经燕麦酰胺C处理的细胞激活了细胞凋亡,因为97%的处理细胞膜联蛋白V染色呈阳性,而91%具有半胱天冬酶-3/7活性,这是细胞凋亡的晚期标志物。
用燕麦酰胺C处理的乳腺癌细胞活力降低,亚G期群体增加,膜联蛋白V和半胱天冬酶活性染色均呈阳性,表明燕麦酰胺C诱导乳腺癌细胞凋亡。这些化合物可能能够作为化学治疗剂,未来的体内研究将对此进行验证。