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凝血剂 Moringa oleifera 凝集素 (cMoL) 对 B16-F10 黑素瘤细胞的细胞毒性。

Cytotoxicity of the coagulant Moringa oleifera lectin (cMoL) to B16-F10 melanoma cells.

机构信息

Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, 50670-901 Recife, Pernambuco, Brazil.

Departamento de Patologia Clínica, Faculdade de Ciências Médicas, Universidade Estadual de Campinas, 13083-887 Campinas, São Paulo, Brazil.

出版信息

Toxicol In Vitro. 2017 Oct;44:94-99. doi: 10.1016/j.tiv.2017.06.019. Epub 2017 Jun 20.

Abstract

Moringa oleifera seeds are used in alternative medicine to treat inflammation, tumors and bacterial and protozoan infections, for example. The seeds contain lectins, which are carbohydrate-binding proteins with several biological properties including cytotoxicity to cancer cells. In this work, we examined the cytotoxicity of the coagulant M. oleifera lectin (cMoL) on B16-F10 murine melanoma cells. cMoL cytotoxic effects were evaluated through trypan blue assay and flow cytometry analysis. Mitochondrial superoxide levels and activation of caspases 3, 8 and 9 were measured. cMoL (1.5-16μM) reduced viability and caused cell death of B16-F10 cells with an IC of 9.72μM. Flow cytometry analysis indicated induction of necrosis and suggested the presence of cells in late apoptosis. Specificity for tumor cells was observed since death of normal human fibroblasts (GN) was not higher than 20% in treatments with cMoL from 1.5 to 16μM. Microscopy images revealed rounded shape and reduction of volume in B16-F10 cells treated with cMoL. cMoL increased mitochondrial ROS production and promoted caspases 3, 8 and 9 activation in B16-F10 cells, indicating the activation of apoptosis-related pathway. In conclusion, this study demonstrates that cMoL is cytotoxic to B16-F10 cells, which stimulates more investigation on the anticancer potential of this lectin.

摘要

辣木籽在替代医学中被用于治疗炎症、肿瘤以及细菌和原生动物感染等疾病。这些种子含有凝集素,这是一种具有多种生物学特性的碳水化合物结合蛋白,包括对癌细胞的细胞毒性。在这项工作中,我们研究了凝固素 M. oleifera 凝集素 (cMoL) 对 B16-F10 鼠黑色素瘤细胞的细胞毒性。通过台盼蓝法和流式细胞术分析评估 cMoL 的细胞毒性作用。测量线粒体超氧化物水平和 caspase-3、8 和 9 的激活。cMoL(1.5-16μM)降低了 B16-F10 细胞的活力并导致其死亡,IC 为 9.72μM。流式细胞术分析表明诱导了坏死,并表明存在晚期凋亡的细胞。观察到肿瘤细胞的特异性,因为在 cMoL 浓度为 1.5 至 16μM 的处理中,正常人类成纤维细胞(GN)的死亡率不高于 20%。显微镜图像显示 cMoL 处理后的 B16-F10 细胞呈圆形且体积减小。cMoL 增加了 B16-F10 细胞中线粒体 ROS 的产生,并促进了 caspase-3、8 和 9 的激活,表明激活了与凋亡相关的途径。总之,这项研究表明 cMoL 对 B16-F10 细胞具有细胞毒性,这激发了对这种凝集素的抗癌潜力的更多研究。

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