Anti-proliferative Effect of Aminoethylated Chitooligosaccharides (below 1 kDa Molecular Weight) in AGS Human Gastric Adenocarcinoma Cells
- Alternative Title
- 아미노에틸화 키토올리고당의 인체위암세포주 증식에 미치는 효과
- Abstract
- In this study, the ability of aminoethylation of chitooligosaccharide (COS) to inhibit the proliferation of AGS human gastric adenocarcinoma cells were evaluated. As aminoderivatized COS, aminoethyl-chitooligosaccharide (AE-COS), dimethyl aminoethyl-chitooligosaccharide (DMAE-COS) and diethyl aminoethyl-chitooligosaccharide (DEAE-COS), were synthesized and confirmed by their IR spectra results in comparison to previous study. Aminoderivatized COS-induced cell death was characterized by cell viability assay, changes in nuclear morphology and changes in cell morphology. According to our results, all aminoderivatized COS significantly induced cell death in AGS gastric cancer cells. Moreover, protein and gene expression levels of important regulators involved in apoptosis pathway such as Caspase 9, Bax, p53 and p21 were examined using RT-PCR and Western blot analysis. Aminoderivatized COS showed dose- and time-dependent inhibition of AGS cancer cell proliferation. Furthermore, anti-apoptotic effects of synthesized COS derivatives were compared with COS. The present results suggest that all three kinds of water-soluble aminoderivatized COS have a promising potential as valuable as cancer chemopreventive agents.
- Issued Date
- 2011
- Awarded Date
- 2011. 2
- Type
- Dissertation
- Keyword
- Biochemistry
- Publisher
- Pukyong National University, Department of Chemistry
- Department
- 대학원 화학과
- Advisor
- Se-Kwon Kim
- Table Of Contents
- 1. Introduction 1
1.1. Chitooligosaccharides 1
1.1.1. Properties of chitin, chitosan and COS 5
1.1.2. Biological activities of COS 6
1.2. Gastric cancer 7
1.3. Apoptosis. 11
1.3.1. Morphology of apoptosis 12
1.3.2. Mechanism of apoptosis. 13
1.4. Research objectives. 18
2. Materials and Methods 19
2.1. Materials 19
2.2. Synthesis of chitoligosaccharide derivatives. 19
2.3. Infrared spectroscopy 20
2.4. Instrumental analysis. 21
2.5. Cell culture 21
2.6. Cell viability assay. 21
2.7. Observation of morphological changes. 22
2.8. Hoechst 33342 staining. 23
2.9. DNA fragmentation. 23
2.10. Fluorescence activated cell sorting. 24
2.11. RNA extraction 25
2.12. Reverse transcription polymerase chain reaction 26
2.13. Protein isolation and Western blot analysis. 30
2.14. Statistical analysis 31
3. Results and Discussion 32
3.1. Synthesis of aminoethylated chitooligosaccharides 32
3.2. Cell viability 39
3.3. Morphological changes and Hoechst 33342 staining 42
3.4. Fluorescence activated cell sorting 48
3.5. DNA fragmentation 53
3.6. Apoptotic effect in gene and protein expression levels 55
4. Summary 62
References 64
- Degree
- Master
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