Smolina Margarita, Goormaghtigh Erik
Laboratory for the Structure and Function of Biological Membranes, Center for Structural Biology and Bioinformatics, Université Libre de Bruxelles (ULB), Bld du Triomphe 2, CP206/02, B-1050 Brussels, Belgium.
Analyst. 2015 Apr 7;140(7):2336-43. doi: 10.1039/c4an01833h.
One current challenge in the field of breast cancer infrared imaging is the identification of carcinoma cell subtypes in the tissue. Neither sequencing nor immunochemistry is currently able to provide a cell by cell thorough classification. The latter is needed to build accurate statistical models capable of recognizing the diversity of breast cancer cell lines that may be present in a tissue section. One possible approach for overcoming this problem is to obtain the IR spectral signature of well-characterized tumor cell lines in culture. Cultures in three-dimensional matrices appear to generate an environment that mimics better the in vivo environment. There are, at present, series of breast cancer cell lines that have been thoroughly characterized in two- and three-dimensional (2D and 3D) cultures by full transcriptomics analyses. In this work, we describe the methods used to grow, to process, and to characterize a triple-negative breast cancer cell line, MDA-MB-231, in 3D laminin-rich extracellular matrix (lrECM) culture and compare it with traditional monolayer cultures and tissue sections. While unsupervised analyses did not completely separate spectra of cells grown in 2D from 3D lrECM cultures, a supervised statistical analysis resulted in an almost perfect separation. When IR spectral responses of epithelial tumor cells from clinical triple-negative breast carcinoma samples were added to these data, a principal component analysis indicated that they cluster closer to the spectra of 3D culture cells than to the spectra of cells grown on a flat plastic substrata. This result is encouraging because of correlating well-characterized cell line features with clinical biopsies.
乳腺癌红外成像领域当前面临的一个挑战是识别组织中的癌细胞亚型。目前,测序和免疫化学都无法逐个细胞地进行全面分类。而这是构建能够识别组织切片中可能存在的乳腺癌细胞系多样性的准确统计模型所必需的。克服这一问题的一种可能方法是获取培养中特征明确的肿瘤细胞系的红外光谱特征。在三维基质中的培养似乎能营造出更接近体内环境的环境。目前,通过全转录组学分析,已有一系列乳腺癌细胞系在二维和三维(2D和3D)培养中得到了充分表征。在这项工作中,我们描述了在富含层粘连蛋白的细胞外基质(lrECM)三维培养中培养、处理和表征三阴性乳腺癌细胞系MDA-MB-231的方法,并将其与传统单层培养和组织切片进行比较。虽然无监督分析未能完全区分二维培养的细胞和三维lrECM培养的细胞的光谱,但有监督统计分析实现了近乎完美的分离。当将临床三阴性乳腺癌样本中上皮肿瘤细胞的红外光谱响应添加到这些数据中时,主成分分析表明,它们与三维培养细胞的光谱聚类比与在平坦塑料基质上生长的细胞的光谱聚类更接近。由于将特征明确的细胞系特征与临床活检相关联,这一结果令人鼓舞。