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Analysis of gene expression in normal and neoplastic keratinocytes

O'Shaughnessy, Ryan Francis Lucas; (2000) Analysis of gene expression in normal and neoplastic keratinocytes. Doctoral thesis (Ph.D), UCL (University College London). Green open access

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The epidermis is a constantly renewing tissue. Cells in the basal layer of the epidermis terminally differentiate and are shed as dead squames. The cells responsible for controlling this constant renewal are known as stem cells. Alterations of these stem cells can lead to neoplasms such as basal cell carcinoma. I used differential hybridisation, a technique that allows the analysis of changes in expression of a large number of genes simultaneously, to find differences in gene expression between basal cell carcinoma tissue and normal skin. Two genes, MRP-14 and 8, were found to be upregulated in basal cell carcinoma. Consistent with the link between expression of these genes and hyperplasia, the epidermis over the basal cell carcinoma expressed high levels of these genes. Improvements I made in the differential hybridisation method allowed elucidation of the differences between stem cells and cells with lower proliferative potential in vitro. Careful analysis revealed no changes in gene expression greater then two fold. One gene, the epidermal fatty acid binding protein, E-FABP, showed higher levels in transit cells. Antibody studies revealed E-FABP expression is reduced in the regions of the epidermis thought to house the stem cells. Finally the expression of a potential marker of stem cells, the melanoma specific chondroitin proteoglycan, MCSP, was examined in vitro and in vivo. Antibody studies revealed expression of the protein only above the dermal papillae of cross sections of epidermis. Fluorescence activated cell sorting revealed a population of basal keratinocytes that are both MSCP positive and express high levels of the β1 integrin, a known stem cell marker.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: Analysis of gene expression in normal and neoplastic keratinocytes
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Thesis digitised by ProQuest.
Keywords: Biological sciences; Epidermis
URI: https://discovery.ucl.ac.uk/id/eprint/10100883
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