KB 구강암세포에서 Angelica decursiva 추출물의 세포사멸 활성 = Apoptotic activity of Angelica decursiva extract in KB oral cancer cells

국문 초록 (Abstract)

우리나라에서 민간요법으로 사용되는 바디나물(Angelica decursiva) 뿌리는 진해제, 진통제, 해열제 또는 거담제의 효능을 가진 것으로 알려져 있다. 그러나 Angelica decursiva의 항암효과에 대해 알려진 바는 거의 없다. 따라서 본 연구에서는 KB 구강암세포에서 Angelica decursiva의 에탄올 추출물(EEAD)에 의한 구강암세포 성장억제 효과와 그 기전을 분석하고자 하였다.
EEAD에 의한 구강암세포 성장억제 효과와 그 기전을 밝히기 위해 KB 구강암세포에서 MTT 분석, DNA fragmentation 분석, immunoblotting 및 caspase 활성분석 등을 시행하였다.
사람정상구강각화세포(HNOK)에서 EEAD는 세포성장의 억제를 유도하지 않았으나 KB 구강암세포에서는 시간과 농도에 의존적으로 세포사멸을 유도하는 것을 확인할 수 있었다. KB 구강암세포에 EEAD를 처리한 실험군에서 procaspase-7, procaspase-9의 proteolytic cleavage 현상과 caspase-3/-7의 활성화를 확인할 수 있었다.
위의 결과들을 종합해보면 Angelica decursiva는 KB 구강암세포에서 apoptosis를 유도하여 암세포 성장을 억제시키는 것으로 생각되며, 본 연구의 결과로 EEAD를 이용한 암세포 성장억제의 하나의 방향을 제시할 수 있을 것으로 사료된다.


증심어 : Angelica decursiva, 세포사, 구강암세포, 항암효과

다국어 초록 (Multilingual Abstract)

The root of Angelica decursiva has been used the home remedy in Korean as an antitussive, an analgesic, an antipyretic, and a expectorant. However, the effects of Angelica decursiva on anti-cancer activity have not been clearly established. The main purpose of this study was to investigate the cytotoxic activity and the mechanism of cell death exhibited by the ethanol extract of Angelica decursiva (EEAD) against KB oral cancer cells.
To determine the effects of EEAD on cell growth in KB oral cancer cells and human normal oral keratinocytes (HNOK), it was examined by MTT assay. In addition, to verify the mechanism of apoptosis, DNA fragmentation analysis, immunoblotting and measuring of caspase activity were performed in KB cells.
EEAD induced the apoptotic cell death in a dose- and a time-dependent manner in KB oral cancer cells, but not in HNOK. Treatment of EEAD significantly promoted proteolytic cleavage of procaspase-7 and -9 with a decrease in the amount of procaspase-7 and -9 in the KB cells. In addition, activation of caspase-3/-7 was detected in living KB oral cancer cells by fluorescence microscopy.
Taken together, these results suggest that EEAD could induce the suppression of cell growth and the promotion of cell apoptosis in KB oral cancer cells, and therefore, that it may have potential properties for anti-cancer drug discovery.

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Key Words: Angelica decursiva, apoptosis, oral cancer cells, anti-cancer activity

목차 (Table of Contents)

• TABLE OF CONTENT
• TABLE OF CONTENT...............................................................................ⅰ
• LIST OF FIGURES..................................................................................... ⅲ
• ABSTRACT.................................................................................................... ⅳ
• I. INTRODUCTION....................................................................................... 1
• II. MATERIALS AND METHODS........................................................ 3
• 1. Materials............................................................................................................................. 3
• 2. Plant material and extract preparation..................................................................... 3
• 3. Cell cultures....................................................................................................................... 3
• 4. Inhibition of cell growth (MTT assay).................................................................... 4
• 5. DNA fragmentation analysis........................................................................................ 5
• 6. Immunoblotting................................................................................................................. 5
• 7. Determination of caspase activation.......................................................................... 6
• 8. Data analysis..................................................................................................................... 6
• Ⅲ. RESULTS................................................................................................... 7
• 1. Cytotoxic activity of EEAD ....................................................................................... 7
• 2. DNA fragmentation assay............................................................................................. 7
• 3. Activation of caspases by EEAD ............................................................................. 7
• Ⅳ. DISCUSSION............................................................................................ 9
• Ⅴ. REFERENCES........................................................................................ 11
• Ⅵ. FIGURE LEGENDS.............................................................................. 17
• Ⅶ. FIGURES.................................................................................................. 19
• ABSTRACT in KOREAN....................................................................... 24