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Access Type

WSU Access

Date of Award

January 2011

Degree Type

Thesis

Degree Name

M.S.

Department

Biomedical Engineering

First Advisor

Gregory Auner

Abstract

Breast cancer is the most common form of cancer in woman in the United States. Current imaging techniques have increased detection rates of breast cancer, specifically ductal carcinoma in situ. However, these imaging techniques are unable to provide information at the molecular level, which is essential in understanding the underlying causes of cancer, the pathogenesis of local recurrence, inter- and intra- cellular changes, and treatment efficacy. Most information on cellular behavior has been based off of fixed samples at specific time points, and thus dynamic processes that occur in between those time points are likely to be missed, especially the subtle changes that occur in the progression of normal breast cells to cancer. We aim to address these issues by combining visual observation and molecular characterization techniques.

A custom built live cell imaging system is used to visually observe normal, pre-malignant, and malignant breast cells growing in real-time for a period of 2.5 days. Cells were grown in three-dimensional cultures to depict true mammary structure. Characterization techniques, such as Raman spectroscopy and X-ray photoelectron spectroscopy (XPS) will be used to detect the subtle dynamic changes that occur as cancer progresses, as well characterize the differences in order to understand the history and aggressive nature of cancerous cells. A trypsinized single cell suspension in phenol free media was used for preliminary Raman and XPS studies.

Preliminary research was also performed to study proteomic changes with Raman spectroscopy. Matrix metalloproteinases are known to play a role in the invasiveness and metastasis of cancer. This may be due in part to its cleavage of α-1-antitrypsin (AAT). This is the first attempt to dynamically visualize proteomic changes that occur in cells.

All three modalities were able to show characteristic differences between normal and cancerous breast cells. Future work will be performed to (a) integrate Raman spectroscopy and live cell imaging for long term real-time characterization of cancer progression and (b) further study of the atomic composition and chemistry of cell with XPS to understand the changes of elemental composition of normal versus cancer.