MDA-MB-231 세포에서 5-nitro-5’-hydroxy- indirubinoxime의 항암 효과

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Purpose: The novel indirubin derivatives 5-nitro-5’-hydroxy- indirubinoxime (5-OH-NIO) was designed and tested for antitumor activity both in vitro and in vivo using breast cancer cell lines, MDA-MB-231 cells.
METHODS: Cell viability was examined by MTT assay. To examine apoptotic cell death, Annexin V/PI staining for flow cytometry and Western blot analysis and DAPI staining were performed. Necrotic cell death was determined by leakage of lactate dehydrogenase (LDH), HMGB1, and PI staining in 5-OH-NIO-treated MDA-MB-231 cells.
Six-week-old male nude mice were inoculated s.c. on the right flank with MDA-MB-231 cells. Indirubin derivatives were directly injected into the tumor every other day. Animals were monitored daily and tumor volume was measured by caliper.
Results: Indirubin derivatives showed potent antiproliferative activity on various human cancer cells. 5-OH-NIO inhibited cells proliferation in a dose dependent manner. The ratio of annexin V-positive cells and cleaved caspase-3/7 was increased by 5-OH-NIO treatment in MDA-MB-231 cells, suggesting that 5-OH-NIO induced cell death in MDA-MB-231 cells may be owing to apoptosis. In addition, LDH secretion and PI staining were detected in MDA-MB-231 cells treated with 5-OH-NIO, showing that 5-OH-NIO also induced necrotic cell death in the MDA-MB-231 cells. The inhibitory effect of 5-OH-NIO on tumor growth was confirmed by in vivo tumor xenograft model. 5-OH-NIO induced significant inhibition of tumor growth in nude mice bearing MDA-MB-231-induced tumors.
Conclusions: These data showed that novel indirubin derivatives 5-OH-NIO arrested the tumor growth by inhibiting cell proliferation and inducing apoptosis and necrosis. These findings provide the potential value of indirubin derivatives as novel candidates for antitumor agents.
Alternative Title
The anti-cancer effect of 5-nitro-5’-hydroxy- indirubinoxime in breast cancer MDA-MB-231 cells
Alternative Author(s)
Seong-Min Kwon
일반대학원 치의생명공학과
Awarded Date
Table Of Contents
목 차 ·············································································· ⅰ
도목차 ·········································································· ⅲ
ABSTRACT ····································································· ⅳ

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

Ⅱ. 재료 및 방법 ·································································· 5
1. 재료 및 항체 ································································ 5
2. 세포 배양 ···································································· 5
3. MTT 실험법 ································································ 5
4. Flow cytometry를 이용한 세포주기 분석 ································ 6
5. Flow cytometry를 이용한 세포사멸 분석 ······························ 6
6. DAPI 염색 ···································································· 6
7. Western blot 분석 ··························································· 7
8. Xenograft model ···························································· 7
9. 자료 분석 ····································································· 7

Ⅲ. 결과 ············································································ 8
1. 5-OH-NIO가 MDA-MB-231 세포 증식에 미치는 영향 ················· 8
2. 5-OH-NIO가 MDA-MB-231 세포주기에 미치는 영향 ··············· 11
3. 5-OH-NIO가 MDA-MB-231 세포사멸에 미치는 영향 ················· 15
4. 5-OH-NIO가 MDA-MB-231 세포괴사에 미치는 영향 ················· 19
5. 5-OH-NIO가 MDA-MB-231 xenograft model에서 종양 억제에 미치는 영향 ············································································ 24

Ⅳ. 고찰 ··········································································· 27

V. 참고문헌 ········································································· 31
조선대학교 대학원
권성민. (2012). MDA-MB-231 세포에서 5-nitro-5’-hydroxy- indirubinoxime의 항암 효과.
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General Graduate School > 4. Theses(Ph.D)
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