González-Fernández María José, Ortea Ignacio, Guil-Guerrero José Luis
Food Technology Division, Agrifood Campus of International Excellence, ceiA3, University of Almería, E-040120 Almería, Spain.
Instituto de Investigación e Innovación Biomédica de Cádiz (INiBICA), Cádiz 11009, Spain.
Toxicol Res (Camb). 2020 Jul 27;9(4):474-483. doi: 10.1093/toxres/tfaa046. eCollection 2020 Jul.
α-Linolenic acid (ALA, 18:3-3) and γ-gamma linolenic acid (GLA, 18:3-6) are polyunsaturated fatty acids (PUFA) that improve the human health. The present study focused on testing the antitumor actions of pure ALA and GLA on the HT-29 human colorectal cancer cell line. Cell viability was checked by MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test, cell membrane damage by the lactate dehydrogenase assay, apoptosis was tested by both caspase-3 activity trial and transmission electron microscopy images, and protein composition was analyzed by quantitative proteomics analysis. MTT test revealed IC values of 230 and 255 μM for ALA and GLA, respectively, at 72 h. After 24 h of incubation, both ALA and GLA induced apoptosis on HT-29 colorectal cancer cells according to the caspase-3 assay and microscopy images. SWATH/MS analysis evidenced that ALA significantly affected the mitochondrial protein import pathway and the citric acid cycle pathway, while GLA did not significantly affect any particular pathway. In summary, both ALA and GLA showed concentration-dependent inhibitory effects on HT-29 cells viability and induced cell death by apoptosis. ALA significantly affected cellular pathways, while GLA does not have specific actions on either pathway. Both -3 and -6 C18 PUFA are bioactive food components useful in the colorectal cancer prevention.
α-亚麻酸(ALA,18:3-3)和γ-亚麻酸(GLA,18:3-6)是对人体健康有益的多不饱和脂肪酸(PUFA)。本研究着重测试纯ALA和GLA对HT-29人结肠癌细胞系的抗肿瘤作用。通过MTT((3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐)试验检测细胞活力,通过乳酸脱氢酶测定检测细胞膜损伤,通过半胱天冬酶-3活性试验和透射电子显微镜图像检测细胞凋亡,并通过定量蛋白质组学分析蛋白质组成。MTT试验显示,在72小时时,ALA和GLA的IC值分别为230和255μM。孵育24小时后,根据半胱天冬酶-3检测和显微镜图像,ALA和GLA均诱导HT-29结肠癌细胞凋亡。SWATH/MS分析证明,ALA显著影响线粒体蛋白质导入途径和柠檬酸循环途径,而GLA对任何特定途径均无显著影响。总之,ALA和GLA均对HT-29细胞活力表现出浓度依赖性抑制作用,并通过凋亡诱导细胞死亡。ALA显著影响细胞途径,而GLA对任一途径均无特定作用。-3和-6 C18 PUFA均为对预防结肠癌有益的生物活性食品成分。