Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Biological Activities of Pharbitis nil and Partial Purification of Anticancer Agent from Its Extract

견우자의 생리활성 분석과 추출물로부터 항암 활성물질의 분리

  • Choi, Hyeun Deok (Department of Microbiology & Immunology, Busan National University School of Medicine) ;
  • Yu, Sun Nyoung (Department of Microbiology & Immunology, Busan National University School of Medicine) ;
  • Park, Sul-Gi (Department of Microbiology & Immunology, Busan National University School of Medicine) ;
  • Kim, Young Wook (Department of Microbiology & Immunology, Busan National University School of Medicine) ;
  • Nam, Hyo Won (Department of Microbiology & Immunology, Busan National University School of Medicine) ;
  • An, Hyun Hee (Department of Microbiology & Immunology, Busan National University School of Medicine) ;
  • Kim, Sang Hun (Department of Microbiology & Immunology, Busan National University School of Medicine) ;
  • Kim, Kwang-Youn (Department of Herbal Formula, Medical Research Center (MRCGHF), College of Oriental Medicine, Daegu Haany University) ;
  • Ahn, Soon Cheol (Department of Microbiology & Immunology, Busan National University School of Medicine)
  • 최현덕 (부산대학교 의학전문대학원 미생물학 및 면역학 교실) ;
  • 유선녕 (부산대학교 의학전문대학원 미생물학 및 면역학 교실) ;
  • 박슬기 (부산대학교 의학전문대학원 미생물학 및 면역학 교실) ;
  • 김영욱 (부산대학교 의학전문대학원 미생물학 및 면역학 교실) ;
  • 남효원 (부산대학교 의학전문대학원 미생물학 및 면역학 교실) ;
  • 안현희 (부산대학교 의학전문대학원 미생물학 및 면역학 교실) ;
  • 김상헌 (부산대학교 의학전문대학원 미생물학 및 면역학 교실) ;
  • 김광연 (대구한의대학교 한의과대학 MRC센터 방제학교실) ;
  • 안순철 (부산대학교 의학전문대학원 미생물학 및 면역학 교실)
  • Received : 2016.12.24
  • Accepted : 2017.02.03
  • Published : 2017.02.28
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Abstract

This study aimed to evaluate several biological activities of Pharbitis nil and to isolate an anticancer agent from its methanol extract. Pharbitis nil seeds were extracted with methanol (PNM). Then, PNM was fractionated into solvent layers such as ethyl acetate fraction (PNE), butanol fraction (PNB), and water fraction (PNW). The biological activities of the fractions were analyzed for tyrosinase inhibition, lipase inhibition, DPPH-free radical scavenging, and cell growth inhibition. PNM showed strong growth inhibition of prostate cancer PC-3 cells. PNM was subjected to Diaion HP-20 and eluted stepwise with 50%, 80%, and 100% methanol. Then, for activity-guided fraction, each fraction was analyzed for growth inhibition of prostate cancer PC-3 cells by using an MTT assay. Because the 100% fraction showed significantly strong inhibitory activity, the fraction was further separated in the reverse phase C18, which was eluted with 80% and 90% methanol. The 90% fraction was further subjected to Sephadex LH-20 using a mobile solvent of 100% methanol. Finally, the compound PN was partially purified for HPLC analysis. PN showed cell growth inhibitory activity and induced the apoptosis and cell cycle arrest of prostate cancer PC-3 cells, as measured by flow cytometry. The results together suggest that Pharbitis nil possesses various biological activities, especially the inhibitory activity for the proliferation of prostate cancer PC-3 cells, suggesting the possibility of its use as an anticancer agent.

견우자는 한의학에서 사하 작용 및 이뇨 작용 등의 치료 목적으로 사용되어 왔다. 본 연구에서는 견우자를 methanol로 추출하고(PNM), 이를 기본으로 ethyl acetate 분획물(PNE), butanol 분획물(PNB), water 분획물(PNW)로 나누었으며 각 분획물을 이용하여 mushroom tyrosinase 저해활성, pancreatic lipase 저해활성, DPPH free radical 소거활성 및 암세포주의 증식 저해활성 등을 조사하였다. 특히, 전립선암 세포주 PC-3 세포에서 강력한 세포 증식 저해활성을 보였으며, 용매 분획물 중, butanol 분획에서 가장 높은 암세포의 증식 저해활성을 보였다. 따라서 견우자의 methanol 추출물을 Diaion HP-20 column chromatography, reverse phase column chromatography, Sephadex LH-20 column chromatography를 순차적으로 실시하여 최종적으로 저해활성이 가장 높은 compound PN을 얻었으며 HPLC를 통해 그 순도를 확인하였다. Compound PN을 이용하여 cell cycle arrest과 annexin V/PI를 측정한 결과, 세포의 sub-G1기를 축적되면서 apoptosis가 유발되어 전립선암 세포주 PC-3 세포의 세포사멸이 유도되는 것을 확인하였다. 따라서 본 연구를 통하여 견우자에 대한 항산화 활성, 미백 활성 등의 효과 뿐 만 아니라 항암 활성에도 활용할 수 있을 것으로 사료된다.

Keywords

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