Tissue-specific microdissection coupled with ProteinChip array technologies: applications in cancer research.
Analysis of whole genomes to monitor specific changes in gene activation or changes in gene copy number due to perturbation has recently become possible using DNA chip technologies. It is now becoming apparent, however, that knowing the genetic sequence encoding a protein is not sufficient to predict the size or biological nature of a protein. This can be particularly important in cancer research where posttranslational modifications of a protein can specifically lead to the disease. To address this area, several proteomic tools have been developed. Currently the most widely used proteomics tool is two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), which can display protein expression patterns to a high degree of resolution. However, 2D-PAGE can be time consuming; the analysis is complicated and, compared with DNA techniques, is not very sensitive. Although some of these problems can be alleviated by using high-quality homogeneous samples, such as those generated using microdissection techniques, the quantity of sample is often limited and may take several days to generate sufficient material for a single 2D-PAGE analysis. As an alternative to 2D-PAGE, a preliminary study using a new technique was used to generate protein expression patterns from either whole tissue extracts or microdissected material. Surface-enhanced laser desorption and ionization allows the retention of proteins on a solid-phase chromatographic surface or ProteinChip Array with direct detection of retained proteins by time-of-flight mass spectrometry. Using this system, we analyzed tumor and normal tissue from head and neck cancer and microdissected melanoma to determine differentially expressed proteins. In particular, comparisons of the protein expression patterns from microdissected normal and tumor tissues indicated several differences, highlighting the importance of extremely defined tissue lysates for protein profiling.
von Eggeling
F
F
Institute of Human Genetics and Anthropology, Jena, Germany. fegg@mit-n.uni-jena.de
Davies
H
H
Lomas
L
L
Fiedler
W
W
Junker
K
K
Claussen
U
U
Ernst
G
G
D015374:Biosensing Techniques; D002457:Cell Extracts / Q000737:chemistry; D004210:Dissection; D006258:Head and Neck Neoplasms / Q000737:chemistry / Q000473:pathology / Q000601:surgery; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008545:Melanoma / Q000737:chemistry / Q000473:pathology / Q000601:surgery; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000032:analysis* / Q000737:chemistry; D009928:Organ Specificity; D020543:Proteome
D002457:Cell Extracts; D009363:Neoplasm Proteins; D020543:Proteome