Centre for Bio-Separation Technolgy (CBST), VIT University, Vellore, Tamil Nadu-632 014, India.
Chem Biol Interact. 2012 Oct 25;200(1):38-44. doi: 10.1016/j.cbi.2012.08.025. Epub 2012 Sep 12.
Medicinal values of mulberry are known to humans from ancient ages. The white mulberry, Morus alba L. is a rich source of many bioactive phytochemicals. Earlier investigations in our laboratory lead to the purification and characterization of an anti-proliferative lectin (MLL) from the leaves of this plant. Further to that, here we have investigated the mechanism of cell death induction by MLL on human breast cancer (MCF-7) and colon cancer (HCT-15) cells. Cells were treated with GI(50) concentration (concentration of lectin required for 50% inhibition of cell growth) of MLL (8.5 μg/ml for MCF-7 and 16 μg/ml for HCT-15) for 24 h to induce cell death. The induction of apoptosis was studied by morphological analysis, DNA fragmentation, apoptotic cell staining and caspase 3 activity assay. Apoptotic cells in sub G0-G1 phase were monitored using flow cytometry. MLL induced significant morphological changes and DNA fragmentation associated with apoptosis in MCF-7 and HCT-15 cells. Positive annexin V and acridine orange/ethidium bromide stained cells indicated apoptosis induction by MLL. Up-regulation of caspase 3 activity was also found in cells treated with MLL. Flow cytometry analysis showed an increase in the percentage of cells in sub G0-G1 phase confirming the MLL induced apoptosis. In conclusion, MLL induced apoptosis in MCF-7 and HCT-15 cells in a caspase dependent manner.
桑树的药用价值自古以来就为人类所熟知。白桑(Morus alba L.)是许多生物活性植物化学物质的丰富来源。我们实验室的早期研究导致从该植物的叶子中纯化和表征了一种具有抗增殖作用的凝集素(MLL)。此外,在这里我们研究了 MLL 对人乳腺癌(MCF-7)和结肠癌细胞(HCT-15)诱导细胞死亡的机制。用 GI(50)浓度(抑制细胞生长 50%所需的凝集素浓度)的 MLL(MCF-7 为 8.5 μg/ml,HCT-15 为 16 μg/ml)处理细胞 24 小时以诱导细胞死亡。通过形态分析、DNA 片段化、凋亡细胞染色和 caspase 3 活性测定研究细胞凋亡的诱导。使用流式细胞术监测亚 G0-G1 期的凋亡细胞。MLL 诱导 MCF-7 和 HCT-15 细胞发生与凋亡相关的显著形态变化和 DNA 片段化。阳性 Annexin V 和吖啶橙/溴化乙锭染色细胞表明 MLL 诱导了细胞凋亡。还发现用 MLL 处理的细胞中 caspase 3 活性上调。流式细胞术分析显示,亚 G0-G1 期细胞的百分比增加,证实了 MLL 诱导的细胞凋亡。总之,MLL 以依赖 caspase 的方式诱导 MCF-7 和 HCT-15 细胞凋亡。