Chemical Composition and Antiproliferative Activity of Supercritical Extract of Immature Citrus Peel in human cervical carcinoma HeLa cells

미성숙 감귤 과피 초임계 추출물의 성분 분석과 자궁암세포 성장억제효능

  • Moon, Jeong Yong (Subtropical Horticulture Research Institute, Jeju National University) ;
  • Song, YeonWoo (Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University) ;
  • Hyun, Ho Bong (Faculty of Biotechnology, College of Applied Life Sciences, Jeju National University) ;
  • KimCho, Somi (Subtropical Horticulture Research Institute, Jeju National University)
  • 문정용 (제주대학교 아열대원예산업연구소) ;
  • 송연우 (제주대학교 생명공학부) ;
  • 현호봉 (제주대학교 생명공학부) ;
  • 김소미 (제주대학교 아열대원예산업연구소)
  • Received : 2015.11.10
  • Accepted : 2015.12.04
  • Published : 2015.12.31


This study was performed to investigate the antiproliferative activities of supercritical extracts from phalsak(Citrus hassaku Hort ex Tanaka) and yeagam(Citrus iyo Hort. ex Tanaka) against human cervical carcinoma HeLa cells and the chemical compositions of the extracts. The anticancer properties of supercritical extracts were demonstrated using the MTT assay and Hoechst 33342 staining and the compositional analyses were conducted by using gas chromatography-mass spectrometry(GC-MS). The peel extracts of both species exhibited similar antiproliferative effect. The antiproliferative activity of the flesh extracts was not detected up to $400{\mu}g/mL$, whereas peel extracts of phalsak and yeagam reduced cell viability with 87.16% and 92.95% at $400{\mu}g/mL$, respectively. There was a dramatic increase of the apoptotic body formation in the cell treated with peel extracts while no apoptotic body formation detected in the cell treated with flesh extracts at 100, $200{\mu}g/mL$. By GC-MS analysis, 27 and 31 kinds of compounds identified in flesh and peel of phalsak, while 27 and 29 kinds of compounds were identified in flesh and peel of yeagam, respectively. 1,1,4,4-Tetramethyl-2-tetralone(20.86%), alloimperatorin(8.15%), limonene(11.23%), and auraptene(7.29%) were major in peel of phalsak, whereas limonene(22.19%), linalool(11.23%), and ${\gamma}$-sitosterol(9.12%) were major in peel of yeagam.

본 연구의 목적은 제주도 재래감귤인 팔삭과 이예감 초임계 추출물에 대한 인간 자궁암 세포인 HeLa 세포에서의 성장억제 효능을 탐색하고, 각 추출물에서의 유효 활성 성분을 분석하는데 있다. HeLa 세포에서의 성장억제 및 세포사멸 효능을 탐색하기 위해 MTT assay와 Hoechst 33342 염색을 수행하였으며, 성분분석은 가스크로마토그래피 질량분석기(GC/MS)를 이용하였다. 두 종류의 미성숙 감귤 과육, 과피 초임계 추출물에 대한 HeLa 세포의 성장 억제 효능을 비교해 본 결과, 과피 추출물은 팔삭과 이예감 모두 비슷한 세포사멸 효능을 나타냈다. 과피 추출물을 처리한 세포에서는 농도 의존적으로 성장률이 억제되었으며, 처리 농도 $400{\mu}g/mL$에서 팔삭과 이예감 추출물이 각각 87.16%와 92.95%의 세포사멸 효능을 나타냈으나, 과육 추출물의 세포 성장 억제 효능은 처리 농도 $400{\mu}g/mL$까지 관찰되지 않았다. Hoechst 33342 염색을 통해 apoptotic body 형성을 현미경으로 관찰한 결과, 100, $200{\mu}g/mL$ 과육 초임계 추출물을 처리한 세포에서는 apoptpotic body를 관찰할 수 없었으나, 동일한 농도의 과피 추출물을 처리한 세포의 경우에서는 apoptotic body가 현저하게 증가하는 것을 확인 할 수 있었다. GC/MS 분석을 통해 미성숙 팔삭 과육과 과피 초임계 추출물에서 각각 27개와 31개의 화합물을 검출하였으며, 미성숙 이예감 과육과 과피 초임계 추출물에서는 각각 27개와 29개의 화합물을 검출하였다. 팔삭 과피 초임계 추출물에는 1,1,4,4-Tetramethyl-2-tetralone(20.86%), alloimperatorin(8.15%), limonene (11.23%), auraptene(7.29%) 등이 주로 함유되어 있었으며, 이예감 과피 초임계 추출물에는 limonene(22.19%), linalool(11.23%), ${\gamma}$-sitosterol(9.12%) 등이 주로 함유되어 있었다.



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