In this thesis, methods to study gene and protein expression in cells and tissues were developed and utilized in combination with protein-specific antibodies, with the overall objective to attain greater understanding of protein function.To analyze protein expression in in vitro cultured cell lines, a cell microarray (CMA) was developed, facilitating antibody-based protein profiling of cell lines using immunohistochemistry (IHC). Staining patterns in cell lines were analyzed using image analysis, developed to automatically identify cells and immunohistochemical staining, providing qualitative and quantitative measurements of protein expression. Quantitative IHC data from CMAs stained with nearly 3000 antibodies was used to evaluate the adequacy of using cell lines as models for cancer tissue. We found that cell lines are homogenous with respect to protein expression profiles, and generally more alike each other, than corresponding cancer cells in vivo. However, we found variability between cell lines in regards to the level of retained tumor phenotypic traits, and identified cell lines with a preserved link to corresponding cancer, suggesting that some cell lines are appropriate model systems for specific tumor types.Specific gene expression patterns were analyzed in vitiligo vulgaris and malignant melanoma. Transcriptional profiling of vitiligo melanocytes revealed dysregulation of genes involved in melanin biosynthesis and melanosome function, thus highlighting some mechanisms possibly involved in the pathogenesis of vitiligo. Two new potential markers for infiltrating malignant melanoma, Syntaxin-7 and Discs large homolog 5, were identified using antibody-based protein profiling of melanoma in a tissue microarray format…
Contentsl
Introduction
Proteomics
Antibody-based proteomics
Immunohistochemistry
Image capture and analysis in immunohistochemistry
Tissue microarray technology
Cell lines
Transcript profiling
DNA microarrays
Quantitative real-time PCR
Diseases of melanocytic origin
Vitiligo vulgaris
Malignant melanoma
Present Investigation
Aims
Results and discussion
Paper I
Paper II
Paper III
Paper IV
Paper V
Conclusions and future perspectives
Populärvetenskaplig sammanfattning
Acknowledgements
References
Author: Stromberg, Sara
Source: Uppsala University Library
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