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Pathogenic mutations and T cell involvement in progression stages of cutaneous melanoma

Optimizing the quantification of molecular genetic changes by digital droplet PCR technology

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Pathogenic mutations and T cell involvement in progression stages of cutaneous melanoma

Optimizing the quantification of molecular genetic changes by digital droplet PCR technology

Open access

Rechten:Alle rechten voorbehouden

Samenvatting

Cutaneous melanoma is a type of skin cancer that develops from benign melanocytes to one of the most aggressive types of skin tumors in human. In cutaneous melanoma progression, the metastasis phase is the most challenging phase to treat. To enable earlier treatment options, a better understanding of the development of this disease is essential. The current knowledge proposes that the stepwise development of melanoma is driven by succession of driver mutations. A common driver in the early stages of melanoma is the V600E mutation in the BRAF oncogene. Though, rogression
from a benign stage to a malignant lesion is not always the case. This might be the result of alternative exon splicing of the BRAF gene. Once the melanoma progresses, an increasing amount of T cells can be observed. Despite the fact that BRAF V600E mutations, splice variants and T cells do play a role in melanoma progression, exact quantitative information is still poorly investigated. In this project, the main goal was to optimize the quantification of molecular genetic changes by digital droplet PCR technology in different progression stages of cutaneous melanoma using formalin-fixed paraffin embedded material derived from dysplastic nevi and melanoma lesions. The central question in this project was: In which quantities are BRAF splice variants and T cells present in the different progression stages of cutaneous melanoma and is the BRAF V600E mutation connected to the
occurrence of splice variants? The results showed that spice variants were present in the BRAF transcripts. However, the exact genomic region where splice variants occurred and the exact connection with the BRAF V600E mutation could not be confirmed. Regarding to T cell quantification, it can be concluded that melanoma contained more T cells than dysplastic nevi. Further research on these subjects is recommended, starting with further optimization of the experiments.

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OrganisatieHZ University of Applied Sciences
OpleidingChemie
AfdelingDomein Technology, Water & Environment
PartnerLeids Universitair Medisch Centrum, Leiden
Datum2018-01-25
TypeBachelor
TaalEngels

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