Date of Award
Master of Science (MS)
Pharmacology and Experimental Therapeutics
Protein kinase C- δ (PKC-δ) acts as a tumor suppressor in skin cancer and is lost in 30% of human Squamous Cell Carcinoma (SCC). This loss is at the transcriptional level and involves the pathway leading to PKC-δ promoter repression after activation of Ras,PI3K,Fyn, and NF-κB. We proposed development of a high throughput, cell-based luciferase reporter assay on Ras transformed keratinocyte cell line, HaCaT-Ras to screen compounds having potential to inhibit this pathway and re-induce PKC-δ expression. First we developed a stable, PKC-δ promoter-reporter transfected HaCaT-Ras cell line. Next, we tested 13 compounds for their abilities to induce PKC-δ promoter activity and identified PP2, and the combination of Bay11-7085 and TPA as positive control compounds. We further optimized and validated the assay on PHERAstar FS to obtain higher Z value. Overall, these studies will be helpful in future for developing a robust high-throughput screening assay of compounds having potential to induce PKC-δ gene expression.
Prajapati, Kushal, "Development of Cell Based Reporter Assay to Measure PKC-Delta Promoter Activity" (2013). Master's Theses. 1862.
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Copyright © 2013 Kushal Prajapati