Tan G, Kaya M, Tevlek A, Sargin I, Baran T
Department of Biology, Faculty of Science and Letters, Aksaray University, TR-68100, Aksaray, Turkey.
Department of Biotechnology and Molecular Biology, Faculty of Science and Letters, Aksaray University, TR-68100, Aksaray, Turkey.
In Vitro Cell Dev Biol Anim. 2018 May;54(5):366-374. doi: 10.1007/s11626-018-0244-8. Epub 2018 Apr 13.
Insects' cuticles have a potential to be evaluated as a chitin source. Especially adults of aquatic insects like mayflies (order Ephemeroptera) swarm in enormous numbers in artificially lit areas while mating in spring and then die by leaving huge amounts of dead insects' bodies. Here in this study, mayfly corpses were harvested and used for production of low MW chitosan. Dried mayfly bodies had 10.21% chitin content; mayfly chitin was converted into chitosan with efficiency rate of 78.43% (deacetylation degree, 84.3%; MW, 3.69 kDa). Cytotoxicity and anti-proliferative activity of mayfly and commercially available shrimp chitosans (low, medium, and high MW) were determined on L929 fibroblast and three different cancer types including HeLa, A549, and WiDr. Apoptosis and necrosis stimulating potential of mayfly and commercial chitosans were also evaluated on A549 and WiDr cells using acridine orange and propidium iodide dual staining to observe morphological changes in nuclei and thus to reveal the predominant cell death mechanism. The effects of chitosans have varied depending on cell types, concentration, and chitosan derivatives. Mayfly and low MW chitosans had a cytotoxic effect at a concentration of 500 μg mL on non-cancer cells. At concentrations below this value (250 μg mL), mayfly and commercial chitosans except high MW one exhibited strong inhibitory activity on cancer cells especially A549 and WiDr cells. Mayfly chitosan induced early and late apoptosis in A549 cells, but late apoptosis and necrosis in WiDr cells. This study suggests that dead bodies of mayflies can be used for production of low MW chitosan with anti-proliferative activity.
昆虫的角质层有潜力被评估为一种几丁质来源。特别是水生昆虫，如蜉蝣（蜉蝣目）的成虫，在春季交配时会大量聚集在人工照明区域，然后死亡，留下大量死虫尸体。在本研究中，收集了蜉蝣尸体并用于生产低分子量壳聚糖。干燥的蜉蝣尸体几丁质含量为10.21%；蜉蝣几丁质转化为壳聚糖的效率为78.43%（脱乙酰度为84.3%；分子量为3.69 kDa）。测定了蜉蝣壳聚糖和市售虾壳聚糖（低、中、高分子量）对L929成纤维细胞以及三种不同癌症类型（包括HeLa、A549和WiDr）的细胞毒性和抗增殖活性。还使用吖啶橙和碘化丙啶双重染色法在A549和WiDr细胞上评估了蜉蝣壳聚糖和商业壳聚糖刺激凋亡和坏死的潜力，以观察细胞核的形态变化，从而揭示主要的细胞死亡机制。壳聚糖的作用因细胞类型、浓度和壳聚糖衍生物而异。蜉蝣壳聚糖和低分子量壳聚糖在浓度为500 μg/mL时对非癌细胞具有细胞毒性。在该值以下的浓度（250 μg/mL）时，除高分子量壳聚糖外，蜉蝣壳聚糖和商业壳聚糖对癌细胞尤其是A549和WiDr细胞表现出强烈的抑制活性。蜉蝣壳聚糖在A549细胞中诱导早期和晚期凋亡，但在WiDr细胞中诱导晚期凋亡和坏死。本研究表明，蜉蝣尸体可用于生产具有抗增殖活性的低分子量壳聚糖。
