Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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The Effect of Red Cabbage (Brassica oleracea L. var. capitata f. rubra) Extract on the Apoptosis in Human Breast Cancer MDA-MB-231 Cells

적양배추 추출물이 인체 유방암 세포 MDA-MB-231의 세포사멸에 미치는 영향

  • Nam, Mi Kyung (Dept. of Food & Nutrition, Duksung Women's University) ;
  • Kang, Keum Jee (Dept. of Food & Nutrition, Duksung Women's University)
  • 남미경 (덕성여자대학교 자연과학대학 식품영양학과) ;
  • 강금지 (덕성여자대학교 자연과학대학 식품영양학과)
  • Received : 2012.08.31
  • Accepted : 2012.11.12
  • Published : 2013.01.31
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Abstract

We investigated the effect of red cabbage extract (RCE) on cell death in MDA-MB-231 human breast cancer cells. Cells were cultured in the presence 1.0, 1.5, and 2.0 mg/mL concentrations of RCE for 24 hours. MTT assays demonstrated that mitochondrial dehydrogenase activities decreased in a dose-dependent manner in cells (p<0.05). In contrast, the proportion of dual staining with Hoechst 33342/ethidium bromide (EtBr) for cell death increased in a dose-dependent manner in cells (p<0.05). Flow cytometry assays revealed that cell death caused by an apoptotic program increased in a dose-dependent (p<0.05). Also, increased ROS accumulation in cells, as revealed by DCF-DA staining, was observed in a dose-dependent fashion (p<0.05). The apoptosis suppressor gene Bcl-2 decreased significantly at the mRNA level. Pro-apoptotic genes Bax and caspase-3, genes that are related to the last stage of apoptosis significantly increased. The Bcl-2/Bax ratio which is an important indicator of apoptosis, was found to have significantly decreased dose dependence. These results taken together indicate that the effect of red cabbage extract induces cell death in MDA-MB-231 human breast cancer cells.

본 연구는 체내 세포배양을 통한 적양배추 추출물의 항유방암 효과를 검정하기 위해 세포활성, 세포사멸, ROS 축적 및 세포사멸 관련 유전자 발현을 분석하였다. 적양배추 추출물을 0.5, 0.75, 1.0, 1.25, 1.5, 1.75 및 2.0 mg/mL의 농도로 24시간 처리하여 세포 생존율(MTT assay)을 분석한 결과 농도 의존적으로 감소되었다(p<0.05). 또한 세포사멸/괴사분석(Hoechst33342/ethidium bromide 염색법), flow cytometry assay, ROS 측정(DCF-DA 염색법) 등의 세포 화학적 방법을 통해 분석한 결과 처리 농도가 증가할수록 세포사멸이 유의적으로 증가하였고, ROS 생성 또한 증가하였다. 특히 2.0 mg/mL의 농도에서는 다른 농도에 비해 유의적으로 높은 사멸율을 나타내었으며, ROS 또한 다량으로 생성되는 것을 관찰할 수 있었다(p<0.05). RT-PCR을 통해 세포사멸관련 유전자인 Bcl-2, Bax, caspase-3의 mRNA 발현 정도를 관찰한 결과 농도 의존적으로 Bcl-2는 유의적으로 감소하였고, Bax와 caspase-3는 유의적으로 증가하였으며, Bcl-2/Bax의 비율은 유의적으로 감소하였다(p<0.05). 이상의 결과로 보아 적양배추 추출물이 인체 유방암세포 MDAMB-231의 세포사멸을 증가시키는 효과가 있음을 알 수 있었다.

Keywords

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