Potcoava Mariana C, Futia Gregory L, Aughenbaugh Jessica, Schlaepfer Isabel R, Gibson Emily A
University of Colorado Denver, Department of Bioengineering, Anschutz Medical Campus, Mail Stop 8607, 12700 East 19th Avenue, Aurora, Colorado 80045.
University of Colorado Denver, Department of Pharmacology, Anschutz Medical Campus, Mail Stop 8303, 12801 East 17th Avenue, Aurora, Colorado 80045.
J Biomed Opt. 2014;19(11):111605. doi: 10.1117/1.JBO.19.11.111605.
Increasing interest in the role of lipids in cancer cell proliferation and resistance to drug therapies has motivated the need to develop better tools for cellular lipid analysis. Quantification of lipids in cells is typically done by destructive chromatography protocols that do not provide spatial information on lipid distribution and prevent dynamic live cell studies. Methods that allow the analysis of lipid content in live cells are therefore of great importance. Using micro-Raman spectroscopy and coherent anti-Stokes Raman scattering (CARS) microscopy, we generated a lipid profile for breast (T47D, MDA-MB-231) and prostate (LNCaP, PC3) cancer cells upon exposure to medroxyprogesterone acetate (MPA) and synthetic androgen R1881. Combining Raman spectra with CARS imaging, we can study the process of hormone-mediated lipogenesis. Our results show that hormone-treated cancer cells T47D and LNCaP have an increased number and size of intracellular lipid droplets and higher degree of saturation than untreated cells. MDA-MB-231 and PC3 cancer cells showed no significant changes upon treatment. Principal component analysis with linear discriminant analysis of the Raman spectra was able to differentiate between cancer cells that were treated with MPA, R1881, and untreated.
人们对脂质在癌细胞增殖和药物治疗耐药性中的作用越来越感兴趣，这促使人们需要开发更好的细胞脂质分析工具。细胞中脂质的定量通常通过破坏性色谱方法进行，这些方法无法提供脂质分布的空间信息，并且阻碍了动态活细胞研究。因此，能够分析活细胞中脂质含量的方法非常重要。利用显微拉曼光谱和相干反斯托克斯拉曼散射（CARS）显微镜，我们在醋酸甲羟孕酮（MPA）和合成雄激素R1881作用下，生成了乳腺癌细胞（T47D、MDA-MB-231）和前列腺癌细胞（LNCaP、PC3）的脂质图谱。将拉曼光谱与CARS成像相结合，我们可以研究激素介导的脂肪生成过程。我们的结果表明，与未处理的细胞相比，经激素处理的癌细胞T47D和LNCaP的细胞内脂滴数量和大小增加，饱和度更高。MDA-MB-231和PC3癌细胞在处理后没有显示出显著变化。对拉曼光谱进行线性判别分析的主成分分析能够区分经MPA、R1881处理的癌细胞和未处理的癌细胞。
