Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Induction of Apoptosis in Human Cancer Cells with Extracts of Taraxacum coreanum, Youngia sonchifolia and Ixeris dentate

흰민들레, 고들빼기, 씀바귀 추출물의 암세포 증식 억제 효과

  • Shin, Seong-Ah (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Lee, Hae-Nim (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Choo, Gang-Sik (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Kim, Hyeong-Jin (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Park, Byung-Kwon (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Kim, Byeong-Soo (Department of Companion and Laboratory Animal Science, Kongju National University) ;
  • Jung, Ji-Youn (Department of Companion and Laboratory Animal Science, Kongju National University)
  • 신성아 (공주대학교 특수동물학과) ;
  • 이해님 (공주대학교 특수동물학과) ;
  • 추강식 (공주대학교 특수동물학과) ;
  • 김형진 (공주대학교 특수동물학과) ;
  • 박병권 (공주대학교 특수동물학과) ;
  • 김병수 (공주대학교 특수동물학과) ;
  • 정지윤 (공주대학교 특수동물학과)
  • Received : 2015.12.24
  • Accepted : 2016.01.13
  • Published : 2016.02.28
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Abstract

This research examined the effects of Compositae extract on the inhibition of proliferation and apoptosis in human breast and human gastric cancer cells. Compositae extracts which is used in the experiment are Taraxacum coreanum (TC), Youngia sonchifolia (YS) and Ixeris dentata (ID). The proliferation of SK-BR-3, MDA-MB-231 and AGS cells were investigated by MTT assay. ID and YS extracts inhibited proliferation of SK-BR-3, MDA-MB-231 and AGS cells in a dose-dependent manner, but TC have barely affected. In addition, the most effective extract was ID. To assess the apoptosis of ID extract, the nuclei of human cancer cells were stained with DAPI solution respectively. Chromatin condensation, indicated apoptosis, was increased in a dose-dependent manner. We investigated change of ID extract-induced apoptosis proteins on human cancer cells by western blot analysis. The level of Bcl-2 decreased, whereas the level of Bax, cleaved-PARP increased in dose-dependent manner compared with non-treatment. Also Bax/Bcl-2 ratio, which is used in clinical indicator of apoptosis, was increased at ID extract treatment group compared with non-treatment. Moreover the Bax/Bcl-2 ratio of MDA-MB-231 cell was significantly increased as against SK-BR-3, AGS cells. These results indicated that ID extract have anti-proliferation effect better than YS or TC, and induced apoptosis in human breast cancer MDA-MB-231 cell better than human breast cancer SK-BR-3 cell, human gastric cancer. Even if further research is needed, ID can be developed as a chemopreventive or therapeutic agent of breast cancer.

본 연구는 인체 유방암세포 SK-BR-3, MDA-MB-231과 위암세포 AGS를 대상으로 국화과 식물 중 흰민들레(Taraxacum coreanum, TC), 고들빼기(Youngia sonchifolia, YS), 씀바귀(Ixeris dentate, ID) 추출물에 의한 증식 억제효과를 비교 한 후, 가장 억제효율이 높은 추출물을 선택하여 apoptosis 유도 효과를 조사하였다. 암세포의 증식 억제효과는 TC 추출물에 의한 영향은 약한 반면, YS와 ID 추출물에 의한 영향은 농도 의존적으로 증가하는 것을 확인하였다. 세 가지 추출물 중 가장 효과가 뛰어난 ID 추출물을 이용하여 추가적으로 농도 설정을 진행한 다음 이후 실험을 진행하였다. ID 추출물에 의한 apoptosis 양성 세포를 확인하기 위해 DAPI assay를 진행한 결과, ID 추출물을 처치한 군에서 apoptotic body와 세포질 응축을 확인하였다. MTT assay와 DAPI staining의 결과를 바탕으로 ID 추출물이 SK-BR-3, MDA-MB-231, AGS 세포에서 apoptosis와 관련한 단백질 발현 양상에 미치는 영향을 확인하기 위해 western blot을 실시하였다. ID 추출물에 의해 SK-BR-3, MDA-MB-231, AGS 세포에서 pro-apoptosis인 Bax 단백질 발현은 농도 의존적으로 증가하였고, anti-apoptosis인 Bcl-2 단백질 발현은 SK-BR-3 세포에서는 발현의 변화가 거의 없었지만, MDA-MB-231과 AGS 세포에서는 감소하였다. 세포사멸의 주요임상지표인 Bax/Bcl-2 ratio는 MDA-MB-231세포에서 가장 유의적으로 증가하였다. 또한, 손상된 DNA를 복구하는 단백질인 PARP의 발현은 감소하면서, cleaved-PARP이 증가하였다. 이러한 결과들을 종합하였을 때, YS와 ID 추출물은 TC 추출물에 비해 암세포의 생존을 효과적으로 억제하였고, 특히 ID 추출물은 낮은 농도에서도 암세포의 생존 억제효과가 우수한 것을 확인하였다. 또한, ID 추출물에 의한 유방암세포 SK-BR-3, MDA-MB-231와 위암세포 AGS의 생존율 억제는 apoptosis 유도를 통해 이루어 지는 것으로 사료되며, 그 중 유방암세포 MDA-MB-231에서 apoptosis 유도 효과가 뛰어난 것을 미루어 보아, 유방암의 종류와 그 특징에 따른 차이를 보이지만 ID는 향후 유방암 예방 및 치료제로 개발 가능성을 제시하며, 추후 지속적인 연구를 통하여 in vivo에서의 ID 추출물의 항암효과에 대해 심도 있는 연구가 이루어 져야 할 것으로 사료된다.

Keywords

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