Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan, Iran.
Department of Pharmaceutical Chemistry, Faculty of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Sciences, Isfahan, Iran.
Iran J Basic Med Sci. 2013 Nov;16(11):1203-8.
OBJECTIVE(S): Regarding the presence of many active biological constituents in Avicennia marina, the present investigation was carried out to study cytotoxic activity of crude methanol leave extract and column chromatographic fractions of A. marina against MDA-MB 231 cell line (human breast cancer cell) and HEK (Human embryonic kidney cell) line.
The anticancer activity of crude methanol extract and sub-fractions were evaluated, using MTT assay. The induction of apoptosis was determined by analyzing DNA fragmentation in breast cancer cells treated with active fraction of crude methanol extract using agarose gel electrophoresis. To investigate molecular mechanism of apoptosis, gene expression levels of p53 and Bcl-2 were measured using quantitative real time PCR.
Fraction 10 was the most active fraction and was detected with HPLC as luteolin. The 50% cell cytotoxic concentration (CC50) of crude methanol extract and luteolin was 250 and 28 µg/ml, respectively. This fraction was found to be an apoptotic agent against MDA-MB 231 cells, which leads to causing DNA fragmentation. The mRNA expression level of Bcl-2 and p53 was significantly decreased and increased respectively in cancer cells treated by luteolin.
The results suggested that Luteolin isolated from Avicennia marina could probably induce apoptosis on breast cancer cell line by the regulation of p53 and Bcl-2 pathways.
鉴于海芒果中存在许多活性生物成分，本研究旨在研究粗甲醇叶提取物和海芒果柱色谱馏分对 MDA-MB 231 细胞系（人乳腺癌细胞）和 HEK（人胚肾细胞）系的细胞毒性活性。
使用 MTT 测定法评估粗甲醇提取物和亚馏分的抗癌活性。通过琼脂糖凝胶电泳分析用粗甲醇提取物的活性部分处理的乳腺癌细胞中的 DNA 片段化来确定细胞凋亡的诱导。为了研究细胞凋亡的分子机制，使用定量实时 PCR 测量 p53 和 Bcl-2 的基因表达水平。
第 10 馏分是最活跃的馏分，并用 HPLC 检测为木犀草素。粗甲醇提取物和木犀草素的 50％细胞细胞毒性浓度（CC50）分别为 250 和 28 µg/ml。该馏分被发现是一种针对 MDA-MB 231 细胞的凋亡剂，可导致 DNA 片段化。用木犀草素处理的癌细胞中 Bcl-2 和 p53 的 mRNA 表达水平分别显著降低和增加。
结果表明，从海芒果中分离出的木犀草素可能通过调节 p53 和 Bcl-2 途径诱导乳腺癌细胞系发生细胞凋亡。
