Date of Award
Master of Science (MS)
College of Science and Mathematics
Thesis Sponsor/Dissertation Chair/Project Chair
Protein-tyrosine kinase, Cellular signal transduction, Neuroblastoma, Cancer--Pathophysiology
The ret proto-oncogene codes for a receptor tyrosine kinase involved in the pathogenesis of numerous developmental defects, particularly those in neural crest-derived structures leading to the regulation of cell proliferation, migration, differentiation and survival during embryogenesis. Constitutive activation or inactivation of the RET receptor results in the cancer syndrome Multiple Endocrine Neoplasia or Hirschsprung's disease, respectively. The importance of RET in these disease states has been well established. However, while many proteins associated with the RET signaling pathway have been identified, the transcriptional changes induced by the wild type receptor, particularly with respect to these signaling proteins in neuroblastoma cells, is less clear. In order to better understand the transcriptional changes that occur in response to RET activation, SK-N-SH neuroblastoma cells were treated with 100 ng/ml GDNF and the RNA isolated. Focusing on the changes in transcription of specific genes that occur in response to the activation of RET, including Etv4, Spryl and Shp2, primers for these genes were designed and the genes characterized using standard RT-PCR and then further analyzed using real-time PGR. Some of these genes promote and control branching in kidney morphology while others, when mutated, are associated with several human diseases. Analyzing the expression levels of each gene in response to the activation of RET will aid in elucidating RET-mediated mechanisms that contribute to these diseases and understanding the role of RET on downstream signaling and cellular function. These findings show quantitatively minimal effect on Spryl, Shp2 and Etv4 expression in response to 4-hour GDNF treatment in neuroblastoma cells by real-time PGR. Because the fold changes observed were not remarkable, the data suggests that expression regulation of these proteins may not be dependent on RET activation. Future studies include analyzing transcriptional changes of these signaling proteins in response to RET activation with longer GDNF treatments or by combination of different ligand stimulation to help further elucidate the possible role of RET on these signaling proteins.
Flores, Elizabeth Yvonne, "A Study of RET Dependent Transcriptional Changes of Cell Signaling Proteins in Neuroblastoma Cells" (2015). Theses, Dissertations and Culminating Projects. 413.