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The Cytotoxicity of Artemisia fukudo Extracts Against HL-60 Cells

큰비쑥(Artemisia fukudo) 추출물의 암세포 증식 억제효과

  • Kim, Kil-Nam (Jeju Biodiversity Research Institute, Jeju Hi-Tech Industry Development Institute) ;
  • Lee, Jung-A (Jeju Biodiversity Research Institute, Jeju Hi-Tech Industry Development Institute) ;
  • Yoon, Weon-Jong (Jeju Biodiversity Research Institute, Jeju Hi-Tech Industry Development Institute) ;
  • Kim, Ji-Young (Jeju Biodiversity Research Institute, Jeju Hi-Tech Industry Development Institute) ;
  • Song, Gwan-Pil (Jeju Biodiversity Research Institute, Jeju Hi-Tech Industry Development Institute) ;
  • Park, Soo-Yeong (Jeju Biodiversity Research Institute, Jeju Hi-Tech Industry Development Institute)
  • 김길남 ((재)제주하이테크산업진흥원 제주생물종다양성연구소) ;
  • 이정아 ((재)제주하이테크산업진흥원 제주생물종다양성연구소) ;
  • 윤원종 ((재)제주하이테크산업진흥원 제주생물종다양성연구소) ;
  • 김지영 ((재)제주하이테크산업진흥원 제주생물종다양성연구소) ;
  • 송관필 ((재)제주하이테크산업진흥원 제주생물종다양성연구소) ;
  • 박수영 ((재)제주하이테크산업진흥원 제주생물종다양성연구소)
  • Published : 2007.07.30

Abstract

The present study describes the cytotoxic effect of Artemisia fukudo extracts. The extract from A. fukudo by 80% ethanol was fractionated with n-hexane, dichloromethane, ethylacetate, and butanol in serial. The cytotoxicity of A. fukudo extracts was examined for its effect on the growth of HL-60 cells by the colorimetric 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide (MTT) assay. In addition, we used the HL-60 cells to see what effects the A. fukudo extracts will have on apoptosis of cancer cells. We checked the cell activity, cell morphologic changes, DNA fragmentation, and DNA content after 24 hr incubation with administering 25 ${\mu}g/mL$ of the A. fukudo extracts. In the treatment of the low concentration of n-hexane and dichloromethane fractions, the survival rate of HL-60 cells is lower than that of the control. The laddering-pattern DNA fragmentation was observed in the treatment with n-hexane and dichloromethane fractions. The DNA content of the cells apoptosis measured as the density of sub-$G_{1}$ hypodiploid cells by flow cytometric analysis. The number of sub-$G_{1}$ hypodiploid cells increased in the treatment with n-hexane and dichloromethane fractions. These fractions obstructed the cell cohesion and caused the blebbing of the cell membrane and fragmentation of the nucleus, both of which are symptoms of apoptosis. These results suggest that A. fukudo has a great potential value as food additives, medicinal supplements for patients with chronic diseases, and preventive measures against cancer.

본 연구는 큰비쑥를 80% EtOH로 추출하고 극성에 따른 용매분획을 실시하여 혈핵암 세포주를 대상으로 분획별에 따른 암세포 증식억제(cytotoxicity) 효과와 이러한 혈액암 세포주에 대한 세포독성 효과가 세포자연사(apoptosis)에 의해 일어나는 것인지, 몇 가지 apoptosis 유도 실험을 통해 확인해 보았다. HL-60 세포에 대한 증식억제 효과는 전반적으로 모든 처리구에서 농도의존적인 세포증식 억제효과가 나타났으며, 특히 헥산과 디클로로메탄 분획물에서 가장 강한 세포성장 억제효과가 나타났다. 이러한 세포성장 억제효과가 apoptosis 유도에 의한 것인지 알아보기 위해 apoptosis 유도에 의한 세포사멸의 특징인 DNA 단편화 현상을 비롯하여 세포형태학적 변화 및 유동세포분석기를 통한 DNA content를 측정한 결과, 큰비쑥 추출물에 의한 암세포 증식억제효과는 apoptosis를 유도하여 세포자연사를 통해 세포의 성장을 억제하는 것으로 확인되었다.

Keywords

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