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Effects of gliotoxin isolated from marine fungus Aspergillus sp. on apoptosis in human cervical cancer cell and human chondrosarcoma cell

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Author(s)
Nguyen Van Tinh
Issued Date
2012
Abstract
Gliotoxin은 항암활성을 포함한 다양한 생리활성을 도출하는 해양 진균 Aspergillus sp.의 이차 대사산물이다. 반면 Gliotoxin의 자궁경부암 (HeLa) 및 연골육종 (SW1353) 세포에 대한 세포사멸 작용기전은 여전히 밝혀지지 않고 있다. 따라서 본 연구에서는 해양 진균에서 분리한 Gliotoxin에 대한 이화학적 특성과 두 종의 암세포주에 대한 세포독성, flow cytometry 분석 및 DNA fragmentation 등 방법으로 세포사멸 효능을 확인하였으며, 아울러 단백질과 유전자 레벨에서의 세포사멸 관련 인자, caspase-3, caspase-8, caspase-9, Bax, Bcl-2, p53 cytochrome c 등을 Western blot 와 RT-PCR 분석방법으로 작용기전을 규명할 수 있었다. 그 결과 Gliotoxin은 두 세포에서 모두 Bcl-2 발현의 감소와 Bax, cytochrome c, caspase-3, caspase-8, caspase-9 발현을 활성화 또는 증가함으로써 세포사멸을 유도시키는 작용기전을 확인하였다.
Alternative Title
해양진균 Aspergillus sp. 로부터 분리된 gliotoxin의 인간 자궁경부암 및 연골육종 세포에서의 세포사멸 효과
Alternative Author(s)
응구엔 반 띤
Affiliation
Chosun University, College of Natural Sciences
Department
일반대학원 해양생명과학
Advisor
Prof. Won-Kyo Jung Ph.D.
Awarded Date
2012-08
Table Of Contents
CONTENTS

CONTENTS i
LIST OF TABLES iv
LIST OF FIGURES v
LIST OF ABBREVIATION vii
초 록 1
I. INTRODUCTION 3
A. Activation and inhibition of apoptosis 3
B. Caspases: the executioners of apoptosis 5
C. Aspergillus sp. and gliotosin 14
II. MATERIALS AND METHOD 19
A. Materials 19
B. Method 19
1. Preparation of Aspergillus sp. 19
2. Isolation of gliotoxin 20
3. Cell line and cell culture 20
4. Cell viability assay 20
5. Nuclear staining with DAPI 21
6. Annexin-FITC V and propidium iodide staining apoptosis tests 21
7. DNA fragmentation assay 22
8. RNA extraction 22
9. Reverse-transcriptase polymerase chain reaction 23
10. Western blot analysis 25
11. Measurement of total protein concentration 25
12. Statistical analysis 25
III. RESULTS AND DISCUSSION 26
A. Extraction of gliotoxin from Aspergillus sp. 26
B. Elucidation of chemical structure of gliotoxin compound 28
C. Cell viability of gliotoxin 35
D. DAPI staining of Hela and SW1353 cells with gliotoxin 37
E. Annexin-FITC V and PI staining apoptosis tests 39
F. DNA fragmentation 42
G. Apoptotic effect in protein and gene expression levels 44
IV. SUMMARY 53
REFERENCES 54
ACKNOWLEDGMENTS 62
Degree
Master
Publisher
Chosun University, College of Natural Sciences
Citation
Nguyen Van Tinh. (2012). Effects of gliotoxin isolated from marine fungus Aspergillus sp. on apoptosis in human cervical cancer cell and human chondrosarcoma cell.
Type
Dissertation
URI
https://oak.chosun.ac.kr/handle/2020.oak/9513
http://chosun.dcollection.net/common/orgView/200000263307
Appears in Collections:
General Graduate School > 3. Theses(Master)
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