Clare Kron

Publication Date

2017

Document Type

Dissertation/Thesis

First Advisor

Bode, Barrie P.

Degree Name

Ph.D. (Doctor of Philosophy)

Legacy Department

Department of Biological Sciences

LCSH

Biology; Molecular biology; Oncology

Abstract

Human hepatocellular carcinoma (HCC) is an increasing threat worldwide due to cirrhosis of the liver developing from viral hepatitis and non-alcoholic steatohepatitis (NASH). Breast cancer is the second leading cause of cancer death in American women, with the most intractable type classified as triple negative. Both cancers depend on glutamine as the major oxidizable substrate for tumor growth, therefore, identification of glutamine transporters and transporter activity is of vital importance in the development of treatment options. It is well documented that ASCT2 is the major glutamine transporter in both cancers but little is known regarding the relative amounts of additional glutamine transporters that operate in these primary and metastatic tumors. Studies using thirteen HCC cell lines investigated eight glutamine transporters via gene expression studies, Western blot assays, and functional analysis. None of the studies, however, incorporated the tumor microenvironment (the stroma), the vital interacting feature of actual tumor growth. In vivo studies were then completed using the EMT6 cell line, a murine mammary carcinoma from a BALB/c laboratory mouse with characteristics of triple negative human breast cancer (TNBC). To distinguish cancer cells from stromal cells, EMT6 was transfected with antibiotic-selective green fluorescent protein (GFP) prior to injection into the mammary fat pads of the genetically identical BALB/c mouse. In vitro results showed that HCC and EMT6 cell lines were glutamine dependent, obtained the majority of glutamine via ASCT2, and used several transporters that were differentially expressed, though transporter activities, cognate mRNA, and protein expression levels were discordant. In vivo mammary carcinoma results showed an increase in all expression levels of transporters, receptors, and enzymes compared to the In vitro EMT6 cell line. EMT6-GFP transfected tumor cells nearly universally induced greater mRNA expression compared to unmodified tumor cells used as a control. The results of this project highlighted the complex and layered regulation between gene expression and physiological activity In vitro , and the changes of mRNA expression In vivo due to both stromal involvement and the effects of antibiotic-selective transfection.

Comments

Advisors: Barrie P. Bode.||Committee members: Mel Duvall; Sherine Elsawa; Elizabeth Gaillard; Wesley Swingley.||Includes bibliographical references.||Includes illustrations.

Extent

xiii, 141 pages

Language

eng

Publisher

Northern Illinois University

Rights Statement

In Copyright

Rights Statement 2

NIU theses are protected by copyright. They may be viewed from Huskie Commons for any purpose, but reproduction or distribution in any format is prohibited without the written permission of the authors.

Media Type

Text

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