Journal of Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Antiproliferative Effect of Extracts, Fractions and Compound from Vitex rotundifolia on Human Cancer Cells

순비기 나무(Vitex rotundifolia) 추출.분획물 및 화합물의 인체 암세포 증식억제 효과

  • Kim, You-Ah (Division of Marine Environment & Bioscience, Korea Maritime University) ;
  • Lee, Jung-Im (Division of Marine Environment & Bioscience, Korea Maritime University) ;
  • Kim, Hae-Jin (Division of Marine Environment & Bioscience, Korea Maritime University) ;
  • Kong, Chang-Suk (Marine Bioprocess Research Center, Pukyong National University) ;
  • Nam, Taek-Jeong (Faculty of Food Science & Biotechnology, Pukyong National University) ;
  • Seo, Young-Wan (Division of Marine Environment & Bioscience, Korea Maritime University)
  • 김유아 (한국해양대학교 해양환경.생명과학부) ;
  • 이정임 (한국해양대학교 해양환경.생명과학부) ;
  • 김해진 (한국해양대학교 해양환경.생명과학부) ;
  • 공창숙 (부경대학교 해양바이오프로세스연구단) ;
  • 남택정 (부경대학교 식품생명공학부) ;
  • 서영완 (한국해양대학교 해양환경.생명과학부)
  • Published : 2009.12.31
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Abstract

Whole plants of Vitex rotundifolia were extracted for 2 days with methylene chloride ($CH_2Cl_2$) followed by extraction of the residue for an additional 2 days. The same procedure was also applied using methanol (MeOH). The two crude extracts were combined and partitioned between $CH_2Cl_2$ and $H_2O$. The organic layer was further partitioned between n-hexane and 85% aq. MeOH, and the aqueous layer was also further fractionated with n-BuOH and $H_2O$, successively. From the 85% aq. MeOH fraction, one compound was isolated through the repeated HPLC. According to the results of physicochemical data including NMR and MS, the chemical structure of the compound was determined as artemetin (1). The antiproliferative effects of the crude extracts, fractions, and compound against HT1080, AGS, MCF-7 and HT-29 human cancer cells were compared with the control by using MTT assay. In the comparative analysis, the 85% aq. MeOH fraction exhibited the strongest antiproliferative effects on human cancer cell lines in a dose-dependent manner (p<0.05). In addition, exposure of compound 1 isolated from 85% aq. MeOH fraction led to strong antiproliferative effect in HT1080 cancer cell lines. These results suggest that the extracts and compound isolated from V. rotundifolia may be used as potential chemopreventive and chemotherapeutic agents.

본 연구에서는 염생식물의 한 종인 순비기나무의 추출 분획물 및 화합을 이용하여 항발암성 및 생체방어 물질로서의 유용성을 검토하고자 4종의 인체 암세포(HT1080, AGS, MCF-7 및 HT-29)에 대한 증식억제 효과를 검토하였다. 추출물에 대한 암세포 증식억제능 측정 결과, 10, 50 및 $100\;{\mu}g/mL$의 시료 처리 농도에서 농도 의존적으로 암세포의 생존율이 감소하는 경향을 확인하였으며, 모든 세포주에 대하여 methanol 추출물 보다 methylene chloride 추출물이 뛰어난 효과를 나타냄을 알 수 있었다. 따라서, 물질의 극성도에 따른 암세포 증식억제능을 확인하고자 추출물을 순차적으로 용매 분획하여 $H_2O$, n-BuOH, 85% aq. MeOH 및 n-hexane의 네 가지 분획층을 얻었으며, 추출물과 동일한 조건하에서 암세포 증식억제 효과를 재검토하였다. 그 결과, 비교적 극성이 덜한 85% aq. MeOH 분획층에서 우수한 암세포 증식억제능이 확인되었다. $100\;{\mu}g/mL$의 농도에서 85% aq. MeOH 분획층의 시료를 처리한 결과 HT1080, AGS, MCF-7 및 HT-29 세포는 각각 93.8, 91.6, 90.9% 그리고 82.7%의 강력한 억제 효과를 나타내었으며, 낮은 농도의 시료($10\;{\mu}g/mL$)를 처리한 결과에서도 HT1080과 AGS 세포에서 주목할 만한 억제효과를 보여 주었다. 생리활성 결과를 바탕으로 85% aq. MeOH 분획층의 분리를 통해 1종의 화합물을 분리할 수 있었으며, artemetin으로 알려진 화합물로 확인되었다. 분리된 화합물의 암세포 증식억제 효과를 확인해 본 결과 HT1080 세포에 선택적으로 효과를 보임을 확인하였으며, 그 효과는 항암제인 doxorubicin에 상응하는 결과였다. 따라서 본 연구를 통해 염생식물 순비기나무의 각종 인체 암세포에 대한 높은 증식억제 효과와 함께 암예방을 위한 기능성 소재로서의 개발 가능성이 확인되었다. 추후 순비기나무의 85% aq. MeOH 분획층 및 분리된 화합물에 대한 집중적인 연구를 통해 새로운 생리활성물질의 개발이 기대되어진다.

Keywords

References

  1. Ahmad VU, Khan MA, Baqai FT, and Tareen RB (1995) Santoflavone, a 5-deoxyflavonoid from Achillea santolina. Phytochemistry 38, 1305-1307 https://doi.org/10.1016/0031-9422(94)00785-R
  2. Doll R (1992) The lessons of life. Cancer Res 52, 2024S-2029S
  3. Hansen MB, Nielsen SE, and Berg K (1989) Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Methods 119, 203-210 https://doi.org/10.1016/0022-1759(89)90397-9
  4. Hern$\acute{a}$ndez MM, Heraso C, Villarreal ML, Vargas-Arispuro I, and Aranda E (1999) Biological activities of crude plant extracts from Vitex trifolia L. (Verbenaceae). J Ethnopharmacol 67, 37-44 https://doi.org/10.1016/S0378-8741(99)00041-0
  5. Hirobe C, Qiao ZS, Takeya K, and Itokawa H (1997) Cytotoxic flavonoids from Vitex agnus-castus. Phytochemistry 46, 521-524 https://doi.org/10.1016/S0031-9422(97)00127-1
  6. Iinuma M, Matsuura S, and Kusuda K (1980) $^{13}C$-Nuclear magnetic resonance (NMR) spectral studies on polysubstituted flavonoids. I. $^{13}C$-NMR spectra of flavones. Chem Pharm Bull 28, 708-716 https://doi.org/10.1248/cpb.28.708
  7. Jo KJ, Yoon MY, Lee MR, Cha MR, and Park HR (2007) The anticancer effect of extracts from Vitex rotundifolia on human colon carcinoma cell lines. J Korean Soc Appl Biol Chem 50, 228-232
  8. Kawaii S, Tomono Y, Katase E, Ogawa K, and Yano M (1999) Antiproliferative activity of flavonoids on several cancer cell lines. Biosci Biotechnol Biochem 63, 896-899 https://doi.org/10.1271/bbb.63.896
  9. Ko WG, Kang TH, Lee SJ, Kim NY, Kim YC, Sohn DH, and Lee BH (2000) Polymethoxyflavonoids from Vitex rotundifolia inhibit proliferation by inducing apoptosis in human myeloid leukemia cells. Food Chem Toxicol 38, 861-865 https://doi.org/10.1016/S0278-6915(00)00079-X
  10. Kong CS, Yeom YR, Lee JI, Kim YA, Lee JS, and Seo Y (2008) Inhibition effects of extracts and its solvent fractions isolated from Limonium tetragonum on growth of human cancer cells. Korean J Biotechnol Bioeng 23, 177-182
  11. Lee YN (2002) In Flora of Korea, p 241, Kyo-Hak Publishing Co. Ltd., Seoul, Korea
  12. Ng TB, Liu F, and Wang HX (2004) The antioxidant effects of aqueous and organic extracts of Panax quinquefolium, Panax notoginseng, Codonopsis pilosula, Pseudostellaria heterophylla and Glehnia littoralis. J Ethnopharmacol 93, 285-288 https://doi.org/10.1016/j.jep.2004.03.040
  13. Ono M, Yanaka T, Yamamoto M, Ito Y, and Nohara T (2002) New diterpenes and norditerpenes from the fruits of Vitex rotundifolia. J Nat Prod 65, 537-541 https://doi.org/10.1021/np0105331
  14. Park EJ and Pezzuto JM (2002) Botanicals in cancer chemoprevention. Cancer Metast Rev 21, 231-255 https://doi.org/10.1023/A:1021254725842
  15. Park JG, Frucht H, LaRocca RV, Bliss DP, Kurita Y, Chen TR, Henslee JG, Trepel JB, Jensen RT, Johnson BE, Bang YJ, Kim JP, and Gazdar AF (1990) Characterization of cell lines established from human gastric carcinoma. Cancer Res 50, 2773-2780
  16. Rahman AU, Ahmed D, Choudhary MI, Turkoz S, and Sener B (1988) Chemical constituents of Buxus sempervirens. Planta Med 54, 173-174 https://doi.org/10.1055/s-2006-962384
  17. Sadhu SK, Okuyama E, Fujimoto H, Ishibashi M, and Yesilada E (2006) Prostaglandin inhibitory and antioxidant components of Cistus laurifolius, a Turkish medicinal plant. J Ethnopharmacol 108, 371-378 https://doi.org/10.1016/j.jep.2006.05.024
  18. Seo Y, Lee HJ, Kim YA, and Park KE (2004) Antioxidative effect of glasswort (Saliconia herbacea) extract from Daebudo. Proceeding of Current Biotechnology and Bioengineering 10, 1-7
  19. Shin TY, Kim SH, Lim JP, Suh ES, Jeong HJ, Kim BD, Park EJ, Hwang WJ, Rye DG, Baek SH, An NH, and Kim HM (2000) Effect of Vitex rotundifolia on immediate-type allergic reaction. J Ethnopharmacol 72, 443-450 https://doi.org/10.1016/S0378-8741(00)00258-0
  20. Shur YJ (2004) Cancer chemopreventive effects of dietrary phytochemicals. J Korean Assoc Cancer Preven 9, 68-83 https://doi.org/10.1038/nrc1189
  21. Stan SD, Kar S, Stoner GD, and Singh SV (2008) Bioactive food components and cancer risk reduction. J Cell Biochem 104, 339-356 https://doi.org/10.1002/jcb.21623
  22. Wang HY, Cai B, Cui CB, Zhang DY, and Yang BF (2005) Vitexicarpin, a flavonoid from Vitex trifolia L., induces apoptosis in K562 cells via mitochondria-controlled apoptotic pathway. Yao Xue Xue Bao 40, 27-31
  23. Yoshioka T, Inokuchi T, Fujioka S, and Kimura Y (2004) Phenolic compounds and flavonoids as plant growth regulators from fruit and leaf of Vitex rotundifolia. Z Naturforsch 59, 509-514
  24. You KM, Son KH, Chang HW, Kang SS, and Kim HP (1998) Vitexicarpin, a flavonoid from the fruits of Vitex rotundifolia, inhibits mouse lymphocyte proliferation and growth of cell lines in vitro. Planta Med 64, 546-550 https://doi.org/10.1055/s-2006-957511

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