생명과학회지 (Journal of Life Science)

  • 제29권3호
  • /
  • Pages.287-294
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  • 2019
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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갯메꽃 추출물과 유기용매 분획물의 암전이 억제 효과

Anti-invasion Effects of Calystegia soldanella Solvent Extracts and Partitioned Fractions on PMA-stimulated Fibrosarcoma Cells

  • 손재민 (한국해양대학교 해양과학기술전문대학원 해양과학기술융합학과) ;
  • 김준세 (한국해양대학교 해양과학기술전문대학원 해양과학기술융합학과) ;
  • 김호준 (한국해양대학교 해양과학기술대학 해양환경.생명과학부) ;
  • 서영완 (한국해양대학교 해양과학기술전문대학원 해양과학기술융합학과)
  • Son, Jaemin (Ocean Science and Technology School, Korea Maritime and Ocean University) ;
  • Kim, Junse (Ocean Science and Technology School, Korea Maritime and Ocean University) ;
  • Kim, Hojun (Division of Marine Bioscience, Korea Maritime and Ocean University) ;
  • Seo, Youngwan (Ocean Science and Technology School, Korea Maritime and Ocean University)
  • 투고 : 2018.10.25
  • 심사 : 2018.11.13
  • 발행 : 2019.03.30
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초록

갯메꽃은 해안 사구에 분포하며 식물로 높은 환경 적응력을 가지고 있으며 갯메꽃은 항산화, 해열, 살균, 이뇨작용 등에 효과가 있는 것으로 알려져 있다. 본 연구에서는 암전이 과정에서 중요한 역할을 한다고 알려진 matrix metalloproteinases (MMPs)의 일종인 MMP-2와 MMP-9의 활성에 대한 갯메꽃의 억제효과를 methylene chloride (MC) 및 methanol (MeOH) 추출물과 조추출물, 유기용매 분획물을 시료로 하여 인체 섬유육종 HT-1080 세포를 이용하여 ELISA (Enzyme-linked immunosorbent), gelatin zymography, Wound healing assay, RT-PCR, western blot 실험을 통해 조사하였다. 갯메꽃의 MC 및 MeOH 추출물, 조추출물과 4가지 유기용매 분획물이 HT-1080 세포에서 MMP-2와 MMP-9 생성 억제 활성을 나타냄을 확인하였다. 그 중에서도 n-hexane, 85% aq.MeOH 분획물이 전반적으로 MMP-2와 MMP-9에 대한 높은 억제활성을 보였다. 이들 결과로부터 85% aq.MeOH과 n-hexane 분획물에 MMP 억제 활성이 높은 물질이 존재할 것으로 추측되어지며, 갯메꽃 추출물 및 분획물이 암침윤 및 전이를 억제하는 효과적인 항암소재로서의 가능성이 있음을 제시한다. 이에 본 연구팀은 85% aq.MeOH과 n-hexane 분획물에 있는 활성물질을 분리하기 위한 연구를 추가적으로 수행할 계획이다.

Calystegia soldanella is distributed in coastal sand dunes and has high environmental adaptability; it is also known to be effective for anti-oxidant, anti-pyretic, anti-septic, and diuretic action. This study investigated the effect of crude extracts and organic solvent fractions of C. soldanella on MMP-2 and MMP-9 expression, MMP activity, and cell mobility in phorbol-12-myristate-13-acetate (PMA)-induced fibrosarcoma HT-1080 cells. C. soldanella was twice extracted, once with methylene chloride (MC) and once with methanol (MeOH). After the MC and MeOH extracts were combined, their suppressive effects on MMP-2 and MMP-9 expression, MMP enzymatic activity, and gene and protein expression were measured by gelatin zymography, enzyme-linked immunosorbent assay, reverse-transcription polymerase chain reaction, and western blot method. Cell mobility for the HT-1080 cells was observed by wound healing assay. The combined crude extracts showed a significant suppressive effects on MMP-2 and MMP-9 expression. To explore active inhibitory elements, the combined extracts were fractionated according to polarity into with n-hexane, 85% aqueous methanol, n-butanol, and water. Across these four solvent fractions, MMP-2 and MMP-9 activity and cell mobility in the HT-1080 cells were all strongly inhibited by the n-hexane fraction. These results suggest that C. soldanella extract and organic solvent fractions could be used as potent MMP inhibitors for effective anti-cancer treatments to suppress cancer invasion and metastasis.

키워드

SMGHBM_2019_v29n3_287_f0001.png 이미지

Fig. 1. Effect of methylene chloride (MC) and methanol (MeOH) extracts from C. soldanella on cell cytotoxicity (A) and the production of MMP-2 (B) and MMP-9 (C) in PMA-stimulated HT-1080 human fibrosarcoma cells.

SMGHBM_2019_v29n3_287_f0002.png 이미지

Fig. 2. Effect of crude extracts from C. soldanella on cell cytotoxicity (A), MMPs expression tested by gelatin zymography (B) and mRNA (C) and protein (D) expression in PMA- stimulated HT-1080 human fibrosarcoma cells.

SMGHBM_2019_v29n3_287_f0003.png 이미지

Fig. 3. Effect of solvent-partitioned fractions from C. soldanella on cell cytotoxicity (A) and the production of MMP-2 (B) and MMP-9 (C) in PMA-stimulated HT-1080 human fibrosarcoma cells.

SMGHBM_2019_v29n3_287_f0004.png 이미지

Fig. 4. Effect of solvent-partitioned fractions from C. soldanella on cell motility and invasion by wound migration assay (A) and gelatin zymography (B) in HT-1080 human fibrosarcoma cells.

SMGHBM_2019_v29n3_287_f0005.png 이미지

Fig. 5. Effect of solvent-partitioned fractions from C. soldanella on MMPs expression in mRNA (A) and protein (B) levels in HT-1080 human fibrosarcoma cells.

Table 1. RT-PCR primer sequence

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