Purification and Characterization of Tumorsphere Formation Inhibitor derived from Saururus chinensis
- Abstract
- Many studies have supported the existence of cancer stem cells (CSCs) as a subpopulation of cancer cells. CSCs are drug-resistant and radiation-resistant cancer cells to be responsible for tumor progress, maintenance and recurrence of cancer, and metastasis. The breast CSCs inhibitor that inhibits cancer stem cell derived from MCF-7 cells was isolated from the aerial parts of Saururus chinensis. The ethyl acetate fraction obtained from methanol extracts exhibited the outstanding inhibitory effect on mammospohere formation. From the ethyl acetate fraction, a new compound named HP-1 was purified through Silica gel column chromatography, ODS open column chromatography, Sephadex LH-20 column chromatography, preparative thin-layer chromatography, and reversed-phase high performance liquid chromatography. The UV spectra of the HP-1 revealed characteristic absorption peaks at 202 and 261 nm. For the first time, we purified CSCs inhibitor from Saururus chinensis. Thus, the isolated compounds exhibited a strong anti-CSCs activity, which may contribute to the interpretation of the pre-pharmacological and clinical effects of S.chinensis.
- Issued Date
- 2016
- Awarded Date
- 2016. 8
- Type
- Dissertation
- Alternative Author(s)
- Sruong, Sreyhak
- Department
- 대학원 생명공학부
- Advisor
- 이동선
- Table Of Contents
- CONTENTS i
LIST OF ABBREVIATIONS . iii
LIST OF FIGURES iv
LIST OF TABLES . v
ABSTRACT . 1
INTRODUCTION . 2
MATERIALS AND METHODS . 5
1. General Experimental Procedures 5
2. Plant Material . 6
3. Extraction and Characterization of HP-1 from S. chinensis . 6
3.1. Preparation of mehanolic crude extracts 8
3.2. Fractionation of the methanolic extracts by means of liquid-liquid partitioning 8
3.3. Isolation and purification of active substances in the bioactive ethyl acetate fraction by bioassay-guided fractionation . 9
3.4. Identification of active components by P. TLC using UV254 nm fluorescence detection . 10
3.5. HPLC analysis . 11
4. Mammosphere formation assay 12
RESULTS 14
1. Purification of active substances from SCE. 14
1.1. The anti-CSCs activity of solvent-partitioning fractions . 14
1.2. The anti-CSCs activity of fractions obtained from Ethyl acetate aqueous 16
1.3. Open ODS column chromatography of the anti-CSCs activity compound from F6 19
1.4. Separation of the anti-CSCs activity compound from 50% of acetonitrile elutes using Sephadex LH-20 column 21
1.5. Isolation of the anti-CSCs activity compound from compounds separted using Preparative TLC . 23
1.6 Purification of active substance in SCE using Preparative HPLC . 26
2. Identification of active substance in SCE . 29
DISCUSSION 32
CONCLUSION 34
REFERENCES 35
ACKNOWLEDGEMENTS . 41
- Degree
- Master
- Publisher
- 제주대학교 대학원
- Citation
- Sreyhak Sruong. (2016). Purification and Characterization of Tumorsphere Formation Inhibitor derived from Saururus chinensis
- Appears in Collections:
- General Graduate School > Biomaterials Science and Technology
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