A549 세포에서 5-nitro-5’-hydroxy-indirubinoxime의 apoptosis과 autophagy 유도 효과

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BACKGROUND: Our group recently demonstrated that 5-nitro-5'-hydroxy-indirubin-3'-oxime (5-OH-NIO), a novel Indirubin-3'-oxime derivative as potent CDK inhibitors. In this study, the effect of 5-OH-NIO was examined in A549 human non-small lung cancer cells.
METHODS: Cell viability was examined by MTT assay. Cell cycle were examined by Flow cytometry and apoptosis was measured using annexin V/PI staining and immunoblot. Autophagy was characterized by the increase in LC3B-II and the formation of autophagosome and acidic vesicular organelles (AVOs).
RESULTS: 5-OH-NIO inhibited the proliferation of A549 cells in a dose-dependent manner. Cell cycle analysis showed the G2/M arrest in 5-OH-NIO-treated cells. 5-OH-NIO increased the number of apoptotic cells by intrinsic apoptotic pathway. 5-OH-NIO also induced autophagy in A549 cells evidenced by the increased levels of LC3 type II expression and the accumulation of AVOs. The inhibition of autophagy enhanced 5-OH-NIO-mediated apoptosis.
CONCLUSIONS: These results suggest that 5-OH-NIO exerts anti-tumor effects by inducing apoptosis and autophagy, and autophagy inhibition enhances 5-OH-NIO-mediated apoptosis in A549 cells.
Alternative Title
Induction of apoptosis and autophagy by 5-nitro-5’-hydroxy-indirubinoxime in human lung cancer A549 cells
Alternative Author(s)
Jong Pil Baek
일반대학원 치의생명공학과
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Table Of Contents
목 차 ·············································································· ⅰ
도목차 ·········································································· ⅲ
ABSTRACT ·····································································

Ⅰ. 서 론 ········································································· 1

Ⅱ. 재료 및 방법 ·································································· 4
1. 재료 및 항체 ································································ 4
2. 세포 배양 ···································································· 4
3. MTT 분석 ·································································· 4
4. Flow cytometry를 이용한 세포주기 분석 ································ 5
5. Flow cytometry를 이용한 세포 사멸 분석 ······························ 5
6. DAPI 염색 ···································································· 5
7. Western blot 분석 ··························································· 5
8. Monodansylcadaverine (MDC) 염색 ······································ 6
9. Acidic vesicular organelles (AVOs) 측정 ······························· 6
10. 자료 분석 ···································································· 6

Ⅲ. 결과 ············································································ 8
1. 5-OH-NIO가 A549 폐암 세포 증식에 미치는 효과 ····················· 8
2. 5-OH-NIO가 A549 폐암 세포 주기에 미치는 효과 ·················· 11
3. A549세포에서 5-OH-NIO에 의한 세포사멸 조사 및 그 기전 연구 ·· 15
4. A549세포에서 5-OH-NIO에 의한 세포자식작용 유도효과 ············· 22
5. A549세포에서 세포 자식작용의 역할 ······································ 27

Ⅳ. 고찰 ··········································································· 31

V. 참고문헌 ········································································· 34
조선대학교 대학원
백종필. (2012). A549 세포에서 5-nitro-5’-hydroxy-indirubinoxime의 apoptosis과 autophagy 유도 효과.
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General Graduate School > 4. Theses(Ph.D)
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