脂质代谢的改变是包括癌症在内的许多疾病的关键因素。因此,研究不饱和和饱和脂肪酸(FAs)对人体稳态的影响对于了解生活方式疾病的发展至关重要。在本文中,我们专注于棕榈(PA)的影响,亚油酸(LA),和二十碳五烯酸(EPA)在人结肠正常(CCD-18Co)和癌症(Caco-2)单细胞上使用拉曼成像和光谱学。拉曼成像的无标记性质使我们能够在不改变内源性细胞代谢的情况下评估FAs动力学。由于拉曼成像能够可视化单细胞亚结构,我们分析了内质网(ER)的化学成分变化,线粒体,脂滴(LD),和补充FA后的细胞核。分析典型的脂质的拉曼谱带强度比,蛋白质,和核酸(I1656/I1444,I1444/I1256,I1444/I750,I1304/I1256)证明,使用拉曼映射,我们可以观察到ER中FAs的代谢途径,负责吸收外源FAs,从头合成,伸长率,和FAs的去饱和,在负责通过FA氧化产生能量的线粒体中,在专门研究细胞脂肪储存的LD中,在细胞核中,脂肪酸通过脂肪酸结合蛋白运输,人类结肠癌发生的生物标志物。膜的分析表明,FAs的摄取有效地改变了该细胞器的化学成分,对洛杉矶的影响最强。光谱学研究已经使用XTT测试完成,这表明对于Caco-2细胞添加LA或EPA降低了它们的活力,对于LA观察到更强的效果,而对于PA观察到相反的效果。对于正常细胞,使用LA或EPA刺激细胞生长的CCD-18Co补充,而PA则产生了相反的影响。
Altered metabolism of lipids is a key factor in many diseases including cancer. Therefore, investigations into the impact of unsaturated and saturated fatty acids (FAs) on human body homeostasis are crucial for understanding the development of lifestyle diseases. In this paper, we focus on the impact of palmitic (PA), linoleic (LA), and eicosapentaenoic (EPA) acids on human colon normal (CCD-18 Co) and cancer (Caco-2) single cells using Raman imaging and spectroscopy. The label-free nature of Raman imaging allowed us to evaluate FAs dynamics without modifying endogenous cellular metabolism. Thanks to the ability of Raman imaging to visualize single-cell substructures, we have analyzed the changes in chemical composition of endoplasmic reticulum (ER), mitochondria, lipid droplets (LDs), and nucleus upon FA supplementation. Analysis of Raman band intensity ratios typical for lipids, proteins, and nucleic acids (I1656/I1444, I1444/I1256, I1444/I750, I1304/I1256) proved that, using Raman mapping, we can observe the metabolic pathways of FAs in ER, which is responsible for the uptake of exogenous FAs, de novo synthesis, elongation, and desaturation of FAs, in mitochondria responsible for energy production via FA oxidation, in LDs specialized in cellular fat storage, and in the nucleus, where FAs are transported via fatty-acid-binding proteins, biomarkers of human colon cancerogenesis. Analysis for membranes showed that the uptake of FAs effectively changed the chemical composition of this organelle, and the strongest effect was noticed for LA. The spectroscopy studies have been completed using XTT tests, which showed that the addition of LA or EPA for Caco-2 cells decreases their viability with a stronger effect observed for LA and the opposite effect observed for PA. For normal cells, CCD-18 Co supplementation using LA or EPA stimulated cells for growing, while PA had the opposite impact.