Master's Theses

Date of Award

2012

Document Type

Thesis

Department

Biomedical Engineering

Keywords

Cytochrome C- Gifp reporter celline, Human Lung cancer, Apoptosis Pathway

Abstract

"Currently chemotherapy and radiotherapy are the most popular methods for cancer treatments after surgery, but lack of specificity and are thus toxic to the healthy human cells. Studies have shown that targeted therapies have been linked to activation of apoptosis signal pathways (cascade of caspases) in cancer cells including intrinsic and extrinsic pathways. One of the major pathways leading to caspase activation involves the release of cytochrome-c from mitochondria. However, in some tumor cells this pathway is inactivated due to the mutation of mitochondria membrane stabilizing proteins (e.g. Bcl-2, Bcl-xL). Therefore, the overall objective of this proposal was to understand the mechanism and the kinetic of cytochrome-c release from the mitochondria, so novel and improved molecular target therapies can be developed against cancers. For this purpose, we have successfully constructed a cytochrome C-GFP reporter cell line using human lung cancer cells. The result from western blot confirmed the presence of the intact cytochrome C-GFP fusion protein in the reporter cell line. We investigated the release kinetics of cytochrome C-GFP in response to three different apoptosis-inducing reagents; TRAIL, Staurosporine, and Tarceva. The result from this study illustrated that the dynamic release of cytochrome C-GFP is dependent on the cell types and doses. We have also developed a system to investigate the anti-cancer drugs response in a sequence of time interval, Microfluidic Duration Device. The microfluidic duration device with the reporter cell line can be further used for preclinical study in the development of combination agents for target therapy."

 
 

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