An Anti-inflammatory Peptide Isolated from Seahorse Hippocampus kuda bleeler inhibits the Invasive Potential of MG-63 Osteosarcoma Cells
- Abstract
- Osteosarcoma is the most common primary malignancy of bone, and patients often develop pulmonary metastasis. The mechanisms underlying osteosarcoma metastasis remain to be elucidated. Recently, anti-inflammatory agents were shown to be useful in the treatment of tumor progression. We previously isolated a natural anti-inflammatory peptide from the seahorse Hippocampus kuda bleeler. Here, we examined the antitumor metastasis activity of this peptide and investigated its mechanism. The peptide significantly inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced invasive migration was associated with reduced expression of matrix metalloproteinases (MMP1 and MMP2). In addition, Rac1 is a member of the Rho family of small GTPases involved in signal transduction pathways that control proliferation, adhesion, and migration of cells during embryonic development and invasiveness of tumor cells. So, we confirmed that TPA stimulation increased intracellular reactive oxygen species (ROS) generation and small GTPase Rac 1 expression, whereas the peptide decreased ROS generation and Rac 1 activation.Taken together, these results suggest that the peptide inhibits invasive migration of MG-63 osteosarcoma cells by inhibiting MMP1 and MMP2 expression through downregulation of Rac 1-ROS signaling.
- Issued Date
- 2013
- Awarded Date
- 2013. 8
- Type
- Dissertation
- Publisher
- 부경대학교
- Affiliation
- 대학원
- Department
- 대학원 화학과
- Advisor
- 박선주
- Table Of Contents
- Table of Contents
Abstract i
Table of Contents …..ii
List of Figures iv
List of Tables vi
List of Abbreviation vii
1. Introduction 1
2. Material and Methods 5
2.1. Materials and chemicals 5
2.2. Preparaiton of the anti-inflammatory peptide 5
2.3. Cell culture 6
2.4. Cell viability assay 7
2.5. Cell invasion assay 7
2.6. Measurement of ROS 8
2.7. Small GTPase Rac1 activity assay 10
2.8. Reverse transcription-polymerization chain reaction (RT-PCR) 10
3. Result and Discussion 15
3.1. An anti-inflammatory peptide from the seahorse dose not affect the viability of MG-63 osteosarcoma cells. 15
3.2. Peptide inhibits TPA-induced invasive migration of MG-63 osteosarcoma cells 19
3.3. Peptide decrease TPA-induced intracellular ROS generation and Rac1 activation in MG-63 cells 22
3.4. Peptide reduces TPA-induced expression of MMPs in MG-63 cells 28
3.5. Peptide increases TPA-induced expression of Collagen Ⅰ in MG-63 cells 31
4. Discussion 33
5. REFERENCES 39
- Degree
- Master
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