DIT Centre for Radiation and Environmental Science (RESC), Focas Research Institute, Dublin Institute of Technology, Kevin Street, Dublin, 8, Ireland.
Analyst. 2010 Dec;135(12):3070-6. doi: 10.1039/c0an00541j. Epub 2010 Oct 7.
The study of the interaction of anticancer drugs with mammalian cells in vitro is important to elucidate the mechanisms of action of the drug on its biological targets. In this context, Raman spectroscopy is a potential candidate for high throughput, non-invasive analysis. To explore this potential, the interaction of cis-diamminedichloroplatinum(II) (cisplatin) with a human lung adenocarcinoma cell line (A549) was investigated using Raman microspectroscopy. The results were correlated with parallel measurements from the MTT cytotoxicity assay, which yielded an IC(50) value of 1.2 ± 0.2 µM. To further confirm the spectral results, Raman spectra were also acquired from DNA extracted from A549 cells exposed to cisplatin and from unexposed controls. Partial least squares (PLS) multivariate regression and PLS Jackknifing were employed to highlight spectral regions which varied in a statistically significant manner with exposure to cisplatin and with the resultant changes in cellular physiology measured by the MTT assay. The results demonstrate the potential of the cellular Raman spectrum to non-invasively elucidate spectral changes that have their origin either in the biochemical interaction of external agents with the cell or its physiological response, allowing the prediction of the cellular response and the identification of the origin of the chemotherapeutic response at a molecular level in the cell.
体外研究抗癌药物与哺乳动物细胞的相互作用对于阐明药物对其生物靶标的作用机制非常重要。在这种情况下,拉曼光谱是一种潜在的高通量、非侵入性分析候选方法。为了探索这种潜力,使用拉曼微光谱研究了顺二氨二氯铂(II)(顺铂)与人肺腺癌细胞系(A549)的相互作用。结果与 MTT 细胞毒性测定的平行测量相关联,得到的 IC50 值为 1.2 ± 0.2 µM。为了进一步证实光谱结果,还从暴露于顺铂的 A549 细胞中提取的 DNA 和未暴露的对照中获取了拉曼光谱。采用偏最小二乘(PLS)多元回归和 PLS 杰克-knifing 来突出与顺铂暴露以及通过 MTT 测定测量的细胞生理变化以统计学显著方式变化的光谱区域。结果表明,细胞拉曼光谱具有非侵入性阐明起源于外部试剂与细胞的生化相互作用或其生理反应的光谱变化的潜力,允许在分子水平上预测细胞反应并识别化疗反应的起源在细胞中。