Date of Defense

5-4-2001

Department

Chemistry

First Advisor

David Scott Reinhold, Department of Chemistry

Second Advisor

Susan R. Stapleton, Department of Chemistry

Keywords

Cancer cells, cell growth, PI 3-Kinase, positive controls, antibody concentrations

Abstract

Normal cell growth takes place as the result of signal transduction pathways within the cell. The activation of enzymes within these pathways can lead to growth and cell division. It has been suggested that irregularities in signal transduction pathways may explain the uncontrolled growth of cancerous cells. One pathway that may play a role in the uncontrolled cell growth in tumor cells is the phosphatidyl 3-kinase (PI 3-K) pathway. Inhibitor studies, performed in the lab, on normal cell lines (L266-C and L266-J) and fibrosarcoma cell lines (HT1080 and VIP:FT) indicated that the aberrant activation of PI 3-Kinase maybe responsible for the uncontrolled growth of the tumorcell lines. Since activation of the PI 3-Kinase results in phosphorylation of protein kinase B(PKB), it was decided to examine the phosphorylated state of PKBin the tumor cell lines and compare it to normal human fibroblast cell lines. Cell lysates were prepared and a determination of the protein content was done for each cell line. Western Blotting was then used to determine the presence of phosphorylated PKB. It was not possible to draw conclusions about the presence of phosphorylated PKB because the positive control did not work. Ideas for further research include changing antibody concentrations and using different positive controls.

Access Setting

Honors Thesis-Campus Only

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