Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Effect of the Solvent Fractions of Zingiber officinale Roscoe against Thrombintreated Tumor Invasion in Human Oral Squamous Carcinoma YD-10B Cells

YD-10B 인간구강암세포주에서 생강 유기용매 분획물의 항산화, 트롬빈억제 및 thrombin에 의해 처리된 암 침윤 및 전이 억제 효과

  • Kim, Eun-Jung (Department of Biomedical Laboratory Science, Sangji University) ;
  • Kim, Jun-Ho (Department of Fine Chemistry and New Materials, Sangji University)
  • 김은정 (상지대학교 임상병리학과) ;
  • 김준호 (상지대학교 정밀화학신소재학과)
  • Received : 2016.06.27
  • Accepted : 2016.08.04
  • Published : 2016.11.30
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Abstract

Oral squamous cell carcinoma (OSCC) is a common malignant tumor in the oral cavity, comprising up to 90% of oral cancer. Oral cancer is characterized by a marked tendency of local invasiveness and is good for early detection and treatment; therefore, it is recognized as a good model for cancer prevention. The present study investigated the antioxidant, thrombin inhibitory, and anti-invasive activities of the solvent fractions of Zingiber officinale Roscoe. Samples were fractionated into hexane, chloroform, ethyl acetate, butanol, and water fractions, and each of these was assayed individually. The water fraction showed the highest extraction yield at 9.79%(w/w). Anti-oxidative activity was analyzed by DPPH assay. Thrombin inhibitory activity was used to analyze thrombin inhibitor assay. Cell viability was detected by the MTS assay. The activity and mRNA expression of MMP-2 and MMP-9 in human oral squamous carcinoma YD-10B cells were examined by zymography and RT-PCR. The antioxidative activities of hexane and water fractions were 92.38% and 92.96%, respectively. In the thrombin inhibitory activity test, water fraction was the highest, with a value of 65.86%. MMP-2/-9 activation was increased in phorbol 12-myristate 13-acetate (PMA)-induced YD-10B cells. MMP-9 activation was increased in thrombin-treated YD-10B cells. In PMA- or thrombin-treated YD-10B cells, the increased mRNA expression and protein activation of MMP-2/-9 were significantly inhibited in the hexane fraction. Therefore, the hexane fraction obtained from a Zingiber officinale Roscoe water extract is a promising therapeutic anti-invasive agent in oral cancer.

구강편평세포암종은 구강 내에서 발생하는 가장 흔한 암의 형태로서, 구강암의 90%이상을 차지한다. 구강암은 국소적인 침윤의 양상을 나타내며 또한 조기진단과 치료가 용이하여 암예방을 위한 유용한 모델로 인정되고 있다. 본 연구에서는 생강 유기용매 분획물의 항산화 활성, 트롬빈억제 및 PMA 또는 thrombin에 의해 유도된 MMP- 2 및 MMP-9활성 억제 효과를 조사하였다. 시료들은 생강 열수 추출물을 헥산(hexane), 클로로포름(chloroform), 에틸 아세테이트(Ethyl acetate), 부탄올(butanol) 및 물($H_2O$)과 같은 용매로 분획화하여 사용하였고, $H_2O$ 분획물의 수득율이 9.79%로 가장 높았다. 항산화 활성은 DPPH assay, 세포 생존율 측정은 MTS assay, 항염증 활성은 마우스 대식세포 Raw 264.7세포에서 NO 생성 그리고 MMP-2 및 MMP-9의 mRNA 발현 및 단백질 활성 억제는 인간구강편평세포암종 YD-10B 세포에서 RT-PCR과 zymography방법을 통해 측정하였다. 본 연구의 결과에 의하면 MMP-2/-9 활성은 PMA에 의해 YD-10B세포에서 증가하였고, thrombin 처리에 의해서는 MMP-9 활성이 유의한 증가를 보였다. YD-10B 세포에서, PMA 또는 thrombin처리 모두에서 hexane 분획물이 MMP-2/-9의 mRNA 발현 및 단백질 활성을 유의하게 억제하였다. 그리고 항산화 활성은 hexane과 $H_2O$ 분획물에서 92.38%와 92.96%로 높게 나타났다. 또한 $H_2O$ 분획물에서 65.86%로 가장 유의하게 트롬빈 억제 활성을 보였다. 그러므로 본 연구에서는 생강 hexane분획물이 구강암의 우수한 암 침윤 및 전이 억제제로서의 개발 가능성을 제시하고 있다.

Keywords

References

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