Robinson Michael A, Graham Daniel J, Morrish Fionnuala, Hockenbery David, Gamble Lara J
National ESCA and Surface Analysis Center for Biomedical Problems, Department of Chemical Engineering, University of Washington, Seattle, Washington 98195.
National ESCA and Surface Analysis Center for Biomedical Problems, Department of Bioengineering, University of Washington, Seattle, Washington 98195.
Biointerphases. 2015 Jun 28;11(2):02A303. doi: 10.1116/1.4929633.
In this work, four triple negative (TN) cell lines, three ER+ and PR+ receptor positive (RP) cell lines, and one ER+, PR+, and HER2+ cell line were chemically distinguished from one another using time-of-flight secondary ion mass spectrometry (ToF-SIMS) and principal component analysis (PCA). PCA scores separation was observed between the individual cell lines within a given classification (TN and RP) and there were distinctly different trends found in the fatty acid and lipid compositions of the two different classifications. These trends indicated that the RP cell lines separated out based on the carbon chain length of the lipids while the TN cell lines showed separation based on cholesterol-related peaks (in the positive ion data). Both cell types separated out by trends in fatty acid chain length and saturation in the negative ions. These chemical differences may be manifestations of unique metabolic processes within each of the different cell lines. Additionally, the HER2+ cell line was distinguished from three other RP cell types as having a unique distribution of fatty acids including anticorrelation to 18-carbon chain fatty acids. As these cell lines could not be grown in the same growth media, a combination of chemical fixation, rinsing, C60 (+) presputtering, and selection of cellular regions-of-interest is also presented as a successful method to acquire ToF-SIMS data from cell lines grown in different media.
在本研究中，使用飞行时间二次离子质谱（ToF-SIMS）和主成分分析（PCA）对四种三阴性（TN）细胞系、三种雌激素受体（ER）和孕激素受体（PR）阳性（RP）细胞系以及一种ER、PR和人表皮生长因子受体2（HER2）阳性细胞系进行了化学区分。在给定分类（TN和RP）内的各个细胞系之间观察到PCA得分分离，并且在两种不同分类的脂肪酸和脂质组成中发现了明显不同的趋势。这些趋势表明，RP细胞系根据脂质的碳链长度分离，而TN细胞系则根据胆固醇相关峰（在正离子数据中）分离。两种细胞类型均通过负离子中脂肪酸链长度和饱和度的趋势分离。这些化学差异可能是每个不同细胞系内独特代谢过程的表现。此外，HER2阳性细胞系与其他三种RP细胞类型不同，其脂肪酸分布独特，包括与18碳链脂肪酸呈反相关。由于这些细胞系不能在相同的生长培养基中生长，本文还介绍了一种化学固定、冲洗、C60（+）预溅射和选择细胞感兴趣区域的组合方法，作为从在不同培养基中生长的细胞系获取ToF-SIMS数据的成功方法。
