Kacar S, Vejselova D, Kutlu H M, Sahinturk V
1 Faculty of Medicine, Department of Histology and Embryology, Eskisehir Osmangazi University, Eskisehir, Turkey.
2 Faculty of Science, Department of Biology, Anadolu University, Eskisehir, Turkey.
Hum Exp Toxicol. 2018 May;37(5):468-474. doi: 10.1177/0960327117712386. Epub 2017 Jun 1.
Acrylamide is a very common compound even reaching up to our daily foods. It has been studied in a wealth of cell lines on which it proved to have various toxic effects. Among these cell lines, human lung adenocarcinoma cell line (A549) is one of that on which acrylamide's toxicity has not been studied well yet.
We intended to determine the half maximal inhibitory concentration (IC) dose of acrylamide and to investigate its cytotoxic, anti-proliferative and apoptotic effects on A549 cells.
We determined the IC dose by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Then, the mode of cell death was evaluated by flow cytometry using Annexin-V fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining. Next, we performed transmission electron microscopy (TEM) and confocal microscopy analyses for morphological alterations and apoptotic indices.
According to the MTT assay results, A549 cell viability decreases proportionally with increasing acrylamide concentrations and IC for A549 was 4.6 mM for 24 h. Annexin-V FITC/PI assay results indicated that acrylamide induces apoptosis in 64% of the A549 cells. TEM and confocal microscopy analyses showed nuclear condensations, fragmentations, cytoskeleton laceration, and membrane blebbing, which are morphological characteristics of apoptosis.
Our research suggests that acrylamide causes cytotoxic, anti-proliferative, and apoptotic effects on A549 cells at 4.6 mM IC dose in 24 h.
丙烯酰胺是一种非常常见的化合物，甚至存在于我们的日常食物中。人们已在大量细胞系中对其进行了研究，结果证明它具有多种毒性作用。在这些细胞系中，人肺腺癌细胞系（A549）是其中之一，丙烯酰胺对该细胞系的毒性尚未得到充分研究。
我们旨在确定丙烯酰胺的半数最大抑制浓度（IC）剂量，并研究其对A549细胞的细胞毒性、抗增殖和凋亡作用。
我们通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑溴盐（MTT）法测定IC剂量。然后，使用膜联蛋白-V异硫氰酸荧光素（FITC）/碘化丙啶（PI）染色，通过流式细胞术评估细胞死亡方式。接下来，我们进行了透射电子显微镜（TEM）和共聚焦显微镜分析，以观察形态学改变和凋亡指数。
根据MTT分析结果，A549细胞活力随丙烯酰胺浓度的增加而成比例降低，A549细胞24小时的IC为4.6 mM。膜联蛋白-V FITC/PI分析结果表明，丙烯酰胺可诱导64%的A549细胞发生凋亡。TEM和共聚焦显微镜分析显示细胞核凝聚、碎片化、细胞骨架撕裂和细胞膜起泡，这些都是凋亡的形态学特征。
我们的研究表明，丙烯酰胺在24小时内以4.6 mM的IC剂量对A549细胞产生细胞毒性、抗增殖和凋亡作用。
