De Guzman Gerard Q, Cagayan Maria Stephanie Fay S, Bagaoisan Daisy-Mae A, Miranda Shiela Dv
Department of Pharmacology and Toxicology, College of Medicine, University of the Philippines Manila.
Institute of Herbal Medicine, National Institutes of Health, University of the Philippines Manila.
Acta Med Philipp. 2023 Nov 24;57(11):41-50. doi: 10.47895/amp.vi0.6631. eCollection 2023.
Breast cancer is the most common cancer among women in the Philippines and about 3 in every 100 Filipina will be diagnosed with breast cancer in their lifetime. There is a need to discover safe, yet inexpensive herbal extracts with potential cytotoxic properties as potential treatment modalities to treat breast cancer.
This study seeks to explore the cytotoxic and apoptotic properties of the ethyl acetate fraction of the defatted crude methanol leaf extract of in MCF-7 breast cancer cell lines.
Screening for flavonoids of the extracts was performed using TLC, total flavonoids, total phenols, FTIR and LC-MS spectroscopy. The hydrogen peroxide and ferric reducing anti-oxidant power were used as substrates to assess anti-oxidative properties of the extracts. The MTT dye viability assay was used to assess the cytotoxic properties of the extracts against MCF-7 cells. Apoptotic properties of the extracts in MCF-7 cells were determined by caspase-3 activation assay, DNA fragmentation patterns and fluorescence microscopy after annexin-V and propidium iodide staining.
The abundance of flavonoids in the ethyl acetate fraction of the crude methanol leaf extract was established by TLC, FTIR, LC-MS/MS, total flavonoid and total phenol analyses. The anti-oxidative properties of this extract was comparable to ascorbic acid. The median inhibitory concentration (IC) of this extract in MCF-7 breast cancer cell lines was 7.2 mcg/mL while doxorubicin registered an IC of 1.2 mcg/mL. At this concentration, the extract was not cytotoxic to normally-dividing breast epithelial cells. Cytotoxicity of the extract was mediated via apoptosis as demonstrated by DNA fragmentation, caspase-3 activation and fluorescence microscopic analyses.
The study shows that the flavonoid-rich ethyl acetate fraction of the crude methanol leaf extract of possesses potent apoptotic and cytotoxic properties against MCF-7 breast cancer cell lines at low concentrations.
乳腺癌是菲律宾女性中最常见的癌症，每100名菲律宾女性中约有3人在其一生中会被诊断出患有乳腺癌。有必要发现具有潜在细胞毒性特性的安全且廉价的草药提取物，作为治疗乳腺癌的潜在治疗方式。
本研究旨在探索去脂粗甲醇叶提取物的乙酸乙酯部分对MCF-7乳腺癌细胞系的细胞毒性和凋亡特性。
使用薄层色谱法（TLC）、总黄酮、总酚、傅里叶变换红外光谱（FTIR）和液相色谱-质谱联用（LC-MS）光谱法对提取物中的黄酮类化合物进行筛选。以过氧化氢和铁还原抗氧化能力作为底物来评估提取物的抗氧化特性。采用MTT染料活力测定法评估提取物对MCF-7细胞的细胞毒性。通过半胱天冬酶-3激活测定、DNA片段化模式以及膜联蛋白-V和碘化丙啶染色后的荧光显微镜检查来确定提取物在MCF-7细胞中的凋亡特性。
通过TLC、FTIR、LC-MS/MS、总黄酮和总酚分析确定了粗甲醇叶提取物的乙酸乙酯部分中黄酮类化合物的含量。该提取物的抗氧化特性与抗坏血酸相当。该提取物在MCF-7乳腺癌细胞系中的半数抑制浓度（IC）为7.2 mcg/mL，而阿霉素的IC为1.2 mcg/mL。在此浓度下，该提取物对正常分裂的乳腺上皮细胞无细胞毒性。DNA片段化、半胱天冬酶-3激活和荧光显微镜分析表明，提取物的细胞毒性是通过凋亡介导的。
该研究表明，粗甲醇叶提取物中富含黄酮类化合物的乙酸乙酯部分在低浓度下对MCF-7乳腺癌细胞系具有强大的凋亡和细胞毒性特性。
