Department of Biochemistry, Biomedical Institute, UNIRIO, 20211-040, Brazil.
Cell and Molecular Biology Graduate Program, UNIRIO, 20211-040, Brazil.
Oxid Med Cell Longev. 2022 Sep 2;2022:2187696. doi: 10.1155/2022/2187696. eCollection 2022.
Bovine lactoferrin (bLf) is a multifunctional protein widely associated with anticancer activity. Prostate cancer is the second most frequent type of cancer worldwide. This study was aimed at evaluating the influence of bLf on cell viability, cell cycle progression, reactive oxygen species (ROS) production, and rate of apoptosis in the human prostate cancer cell line (DU-145). MTT assay and trypan blue exclusion were used to analyze cell viability. Morphological changes were analyzed through optical microscopy after 24 h and 48 h of bLf treatment. FITC-bLf internalization and cellular damage were observed within 24 h by confocal fluorescence microscopy. Cell cycle analyses were performed by flow cytometry and propidium iodide. For caspases 3/7 activation and reactive oxygen species production evaluation, cells were live-imaged using the high-throughput system Operetta. The cell viability assays demonstrated that bLf induces cell death and morphological changes after 24 h and 48 h of treatment compared to control on DU-145 cells. The bLf internalization was detected in DU-145 cells, G-phase arrest of the cell cycle, caspase 3/7 activation, and increased oxidative stress on bLf-treated cells. Our data support that bLf has an important anticancer activity, thus offering new perspectives in preventing and treating prostate cancer.
牛乳铁蛋白(bLf)是一种多功能蛋白,广泛与抗癌活性相关。前列腺癌是全球第二常见的癌症类型。本研究旨在评估 bLf 对人前列腺癌细胞系(DU-145)活力、细胞周期进程、活性氧(ROS)产生和细胞凋亡率的影响。MTT 检测和台盼蓝排斥试验用于分析细胞活力。bLf 处理 24 h 和 48 h 后通过光学显微镜分析形态变化。24 h 内通过共聚焦荧光显微镜观察 FITC-bLf 内化和细胞损伤。通过流式细胞术和碘化丙啶进行细胞周期分析。为了评估 caspase 3/7 激活和活性氧产生,使用高通量系统 Operetta 对活细胞进行成像。与对照组相比,细胞活力检测显示 bLf 在处理 24 h 和 48 h 后诱导 DU-145 细胞死亡和形态变化。在 DU-145 细胞中检测到 bLf 内化、细胞周期 G 期阻滞、caspase 3/7 激活和 bLf 处理细胞中氧化应激增加。我们的数据支持 bLf 具有重要的抗癌活性,从而为预防和治疗前列腺癌提供了新的视角。