PUKYONG

GUIDE OAK 국가리포지터리 부경대학교

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Abstract: Gastric cancer is the second most common cause of cancer-related deaths in the world. In this study, a bioactive derivative of chitooligosaccharides, named gallic acid-grafted-chitooligosaccharides (G-COS), was evaluated for its capabilities against the proliferation of AGS human gastric carcinoma cell line. It was found that G-COS treatment caused significant inhibition on gastric cancer cell growth at concentration of 200 and 400 µg/ml. The inhibitory effect of G-COS was evidenced via inducing apoptosis. G-COS-induced cell death was identified by cell viability assay, changes in cell and nuclear morphology, DNA fragmentation, apoptosis analysis and cell cycle analysis. Notably, G-COS-induced apoptosis was related to the increase in the expression of p53, p21, Bax, cytochrome c, caspase (-9 and -3), cleaved PARP, and the decrease in the activation of Bcl-2, p-IκB-α and NF-κB (p50 and p65). These findings indicate that G-COS has a promising potential to be applied in the treatment of gastric cancer as cancer chemopreventive agents.

URI: https://repository.pknu.ac.kr:8443/handle/2021.oak/12917
http://pknu.dcollection.net/jsp/common/DcLoOrgPer.jsp?sItemId=000002235132

Table Of Contents: Abstract i
Table of Contents ii
List of Tables v
List of Figures vi
List of Abbreviations vii
1. Introduction 1
1.1. Chitin, chitosan and chitooligosaccharides 1
1.2. Gallic acid 5
1.3. Gastric cancer 7
1.4. Apoptosis 9
1.4.1. Morphology of apoptosis 9
1.4.2. Pathways of apoptosis 10
2. Materials and methods 13
2.1. Materials 13
2.2. Synthesis of G-COS 13
2.3. Cell culture 14
2.4. Cell viability 14
2.5. Morphological analysis and Hoechst staining 15
2.6. DNA fragmentation 16
2.7. Cell cycle analysis 17
2.8. Apoptosis analysis 17
2.9. Western blot analysis 18
2.10. RNA extraction and reverse transcription-polymerase chain reaction (RT-PCR) analysis 19
2.11. Statistical analysis 19
3. Results and Discussion 21
3.1. Structural characterization of G-COS 21
3.2. Cell viability 24
3.3. Morphological changes and Hoechst 33342 staining 26
3.4. DNA fragmentation 28
3.5. Analysis of cell cycle distribution 30
3.6. Analysis of apoptosis 32
3.7. Effects of G-COS on expression of gene and proteins related to apoptosis 34
3.7.1. Effect of G-COS on p53 signaling pathway 34
3.7.2. Effect of G-COS on Bcl-2 family 36
3.7.3. Effect of G-COS on Caspases 39
3.7.4. Effect of G-COS on NF- κB 41
4. Summary and Conclusion 44
References 46
Acknowledgement 57

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