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In vitrocharacterization of lectin-induced alterations on the proliferative activity of three human melanoma cell lines

机译:凝集素诱导的三种人黑色素瘤细胞系增殖活性改变的体外表征

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Lectin binding is known to be able to elicit signalling events relevant for various aspects of cell physiology. The influence of lectin binding on melanoma cells remains relatively unexplored. The aim of our study was to investigate the in vitro effects of five plant lectins, namely peanut (PNA), wheat germ (WGA), concanavalin A (Con-A), Griffonia simplicifolia (GSA-IA4) and Phaseolus vulgaris (PHA-L) agglutinins, on the cell proliferation of melanoma cell lines (SK-MEL-28, HT-144 and C32) cultured in media supplemented with either 10 or 1 fetal calf serum (FCS). Cell proliferation was assessed by means of the tetrazolium derivative reduction (MTT) assay. Four lectin concentrations were tested, namely 0.05, 0.5, 5 and 50 ug/ml, in four experimental settings, namely 1, 3, 5 and 7 days after the addition of each lectin to the culture media. Determination of the cell gain compartment (percentage of cells in the S and G2 phases of the cell cycle) was done by means of digital cell image analysis assessed on Feulgen-stained nuclei. Our results demonstrated that of the five lectins under study, four had a globally significant dose-dependent toxic effect on melanoma cell proliferation. The fifth lectin, PNA, had a significant stimulatory effect on the C32 cell line. Low doses of lectins may produce a transient increase in cell proliferation. Increasing the FCS from 1 to 10 in the culture media significantly antagonized lectin-induced toxicity in the three cell lines. The cell kinetics measurements showed that the inhibition of cell growth was merely due to cell death. The present data strongly suggest that some lectins might influence the proliferation of melanoma cells. In addition, because lectins are present in our diet and are able to pass into the systemic circulation, we speculate that lectins may exert an influence on melanoma growth under clinical conditions.
机译:已知凝集素结合能够引发与细胞生理学各个方面相关的信号传导事件。凝集素结合对黑色素瘤细胞的影响仍相对未被探索。本研究的目的是研究五种植物凝集素的体外影响,即花生 (PNA)、小麦胚芽 (WGA)、刀豆蛋白 A (Con-A)、Griffonia simplicifolia (GSA-IA4) 和菜豆 (PHA-L) 凝集素,对在补充有 10% 或 1% 胎牛血清 (FCS) 的培养基中培养的黑色素瘤细胞系(SK-MEL-28、HT-144 和 C32)的细胞增殖。通过四唑衍生物还原 (MTT) 测定法评估细胞增殖。在四个实验环境中测试四种凝集素浓度,即 0.05、0.5、5 和 50 ug/ml,即在将每种凝集素添加到培养基后 1、3、5 和 7 天。通过对Feulgen染色的细胞核进行数字细胞图像分析来确定细胞增益区室(细胞周期的S期和G2期细胞百分比)。我们的结果表明,在所研究的五种凝集素中,有四种对黑色素瘤细胞增殖具有全球显着的剂量依赖性毒性作用。第五种凝集素PNA对C32细胞系有显著的刺激作用。低剂量的凝集素可能会引起细胞增殖的短暂增加。在培养基中将FCS从1%提高到10%可显著拮抗凝集素诱导的三种细胞系毒性。细胞动力学测量表明,细胞生长的抑制仅仅是由于细胞死亡。目前的数据强烈表明,一些凝集素可能会影响黑色素瘤细胞的增殖。此外,由于凝集素存在于我们的饮食中并且能够进入体循环,我们推测凝集素可能会在临床条件下对黑色素瘤的生长产生影响。

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