Core Unit Chip Application, Institute of Human Genetics, University Hospital Jena, Jena, Germany.
Diagn Pathol. 2010 Jan 28;5:10. doi: 10.1186/1746-1596-5-10.
In carcinoma tissues, genetic and metabolic changes not only occur at the tumor cell level, but also in the surrounding stroma. This carcinoma-reactive stromal tissue is heterogeneous and consists e.g. of non-epithelial cells such as fibroblasts or fibrocytes, inflammatory cells and vasculature-related cells, which promote carcinoma growth and progression of carcinomas. Nevertheless, there is just little knowledge about the proteomic changes from normal connective tissue to tumor stroma. In the present study, we acquired and analysed specific protein patterns of small stromal sections surrounding head and neck cell complexes in comparison to normal subepithelial connective tissue. To gain defined stromal areas we used laser-based tissue microdissection. Because these stromal areas are limited in size we established the highly sensitive 'tissue on chip based mass spectrometry' (toc-MS). Therefore, the dissected areas were directly transferred to chromatographic arrays and the proteomic profiles were subsequently analysed with mass spectrometry. At least 100 cells were needed for an adequate spectrum. The locating of differentially expressed proteins enables a precise separation of normal and tumor stroma. The newly described toc-MS technology allows an initial insight into proteomic differences between small numbers of exactly defined cells from normal and tumor stroma.
在癌组织中，遗传和代谢变化不仅发生在肿瘤细胞水平，也发生在周围基质中。这种癌反应性基质组织是异质的，例如由成纤维细胞或纤维母细胞等非上皮细胞、炎症细胞和与脉管系统相关的细胞组成，这些细胞促进癌的生长和癌的进展。然而，对于从正常结缔组织到肿瘤基质的蛋白质组变化，人们知之甚少。在本研究中，我们获得并分析了头颈部细胞复合体周围的小基质切片的特定蛋白质图谱，与正常上皮下结缔组织进行了比较。为了获得明确的基质区域，我们使用了基于激光的组织微切割。由于这些基质区域的大小有限，我们建立了高度敏感的“基于组织的芯片质谱”（toc-MS）。因此，切割的区域被直接转移到色谱阵列上，随后用质谱分析蛋白质组图谱。至少需要 100 个细胞才能获得足够的光谱。差异表达蛋白的定位可以精确地区分正常和肿瘤基质。新描述的 toc-MS 技术允许初步了解正常和肿瘤基质中少量精确定义的细胞之间的蛋白质组差异。
