Date of Award

2011

Degree Type

Thesis

Degree Name

Master of Science

Program

Medical Biophysics

Supervisor

Dr. John D. Lewis

Abstract

Metastatic disease, or the migration of cancer cells from the primary tumor to distant locations in the body, contributes to over 90% of cancer mortalities. Migration is a requirement of metastasis and involves the detachment of cancer cells from the primary tumour in vivo, followed by invasion of the cell into the surrounding stromal tissue. Tumour cells that are migration-deficient are incapable of detaching from the primary tumour and exhibit compact phenotypes in the chicken embryo model. Based on these findings, it was hypothesized that mediators of migration could be identified using an RNAi genomic library and screening for compact tumour phenotypes in the chicken embryo model. It was also postulated that knockdown of two proteins known to be involved in migration, rhoA and cortactin, would prevent migration of human epidermoid carcinoma (HEp3) cells in vivo, serving as a positive control and proof-of-principle for the RNAi screen. Results of this study identify rhoA and cortactin as positive regulators of migration, both in vitro and in vivo, and demonstrate the feasibility of the RNAi screen. Furthermore, execution of an RNAi screen, covering 5000 human genes, identified three novel mediators of tumour cell migration: MESCD1, KIF3B and ARHGAP12.

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