Autocrine Stimulation Of Melanoma Causes Mmp-2 Upregulation And Subsequent Migration

Open Access
Author:
Khristov, Todor R
Area of Honors:
Bioengineering
Degree:
Bachelor of Science
Document Type:
Thesis
Thesis Supervisors:
  • Cheng Dong, Thesis Supervisor
  • Peter J Butler, Honors Advisor
Keywords:
  • melanoma
  • tumor
  • extravasation
  • matrix metalloproteinase
Abstract:
The goal of this work is to investigate the pathway behind the up-regulation and expression of Matrix Metalloproteinases (MMPs) as a result of autocrine melanoma stimulation. Certain chemokines, such as Monocyte Chemotactic Protein-1 (MCP-1) and Interleukin-8 (IL-8) are characteristic of metastatic melanomas and have previously been implicated to be involved in MMP-2 up-regulation and activation. It is the pathways involving these proteins that are the focus of our research. In order to study the effect of MMPs on melanoma extravasation through the extracellular matrix, an assay was developed, which involved the extract from murine Engelbreth-Holm-Swarm sarcoma (Matrigel), to simulate the extracellular matrix. An optimal Matrigel volume was determined to allow monitoring the changes in melanoma migration as a result of inhibition of certain steps in the MMP activation pathway. The highly metastatic melanoma cell line (Lu1205) was used in the experiments. The study focused on the synergistic effect of MCP-1 and IL-8 on melanoma migration through the extracellular matrix. It was shown that blocking MCP-1 and IL-8 reduced the migration and invasion rate of Lu1205 cells. Additionally, it was shown that MMP-2 plays a crucial role in melanoma extracellular matrix digestion. In order to account for incomplete migration of Lu1205 through the Matrigel, a modified flow chamber was designed.