This thesis focuses on the study of heparanase which is an enzyme involved as a molecular determinant of cancer metastasis. The purpose of this study was to first quantitate heparanase mRNA levels in both normal and tumorigenic samples from the same tissue specimen. Laser capture microdissection was used in the isolation and extraction of melanoma cell populations from normal tissue. There was a 29 fold upregulation of heparanase expression, detected by real-time PCR, in metastatic melanoma of the lung in comparison to normal lung tissue in mice. Immunohistochemistry (IHC) showed stronger staining in human metastatic melanoma when compared to primary melanoma tumors. IHC also showed a propensity for darker heparanase staining around blood vessels and vascular regions. These results further emphasized the importance of heparanase in invasive and angiogenic mechanisms in melanoma. Once heparanase was determined to be upregulated in melanoma tissue in vivo both at the mRNA and protein level, the next part of this thesis was directed to the development of an orthotopic brain slice model. A novel model that would provide a relatively efficient way to study the biological relevance and mechanisms involved in the invasive process of brain metastatic melanoma and the role that heparanase plays in this invasive process. We showed that this model could be used to determine invasion into brain tissue at both qualitative and quantitative levels. We showed that HPSE-1 augmented invasion of brain metastatic melanoma cells into brain tissue. We also showed that melanoma cells show a time dependent expression of heparanase while invading into brain tissue. Thus, we showed that heparanase is involved in cancer metastasis development and could have important implications in the development of potential drugs aimed to combat cancer metastasis.
| Identifer | oai:union.ndltd.org:LSU/oai:etd.lsu.edu:etd-07132005-134343 |
| Date | 14 July 2005 |
| Creators | Murry, Brian Paul |
| Contributors | William G. Henk, Steven A. Barker, Dario Marchetti |
| Publisher | LSU |
| Source Sets | Louisiana State University |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.lsu.edu/docs/available/etd-07132005-134343/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached herein a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to LSU or its agents the non-exclusive license to archive and make accessible, under the conditions specified below and in appropriate University policies, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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