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PMA로 자극된 HT-1080 세포에서 염생식물 광나무의 Matrix metalloproteinases 활성 억제효과
김준세(Junse Kim),공창숙(Chang-Suk Kong),문기환(Ki Hwan Moon),서영완(Youngwan Seo) 한국생물공학회 2020 KSBB Journal Vol.35 No.1
Matrix metalloproteinases (MMPs) are the zincdependent endopeptidases that cause the degradation and remodeling of extracellular matrix. They are associated with various physiological and pathological processes. Among the MMPs, gelatinases, MMP-2 and MMP-9, play a major role in the metastasis and invasion of cancer or tumor cells. In this study, the inhibitory effect of the halophyte Ligustrum Japonicum on MMP-2 and MMP-9 activity in phorbol-12-myristate- 13-acetate (PMA)-stimulated HT-1080 cells was estimated by using enzyme-linked immunosorbent assay (ELISA), gelatin zymography, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assay. Also, the effect of L. japonicum on HT-1080 cell migration was investigated by using wound-healing assay. The dried L. japonicum was extracted with dichloromethane (CH₂Cl2₂) and methanol (MeOH), and the combined crude extracts were fractionated according to the solvent polarity to obtain the n-hexane, 85% aqueous methanol (85% aq.MeOH), n-butanol (n-BuOH) and water fractions. Of the fractions, 85% aq.MeOH fraction most effectively suppressed the enzymatic activity, cell mobility, and mRNA and protein expression of MMP-2 and MMP-9. Therefore, L. japonicum will be used as a valuable source of potential anti-invasive agents.
인간 섬유육종세포에서 비쑥 추출물과 유기용매 분획물의 암전이 억제 효과
김준세(Junse Kim),공창숙(Chang-Suk Kong),심현보(Hyun-Bo Sim),서영완(Youngwan Seo) 한국생명과학회 2021 생명과학회지 Vol.31 No.12
염생식물인 비쑥은 식용이 가능한 약용식물로서 살충, 항염, 항콜레스테롤, 해열, 항균 활성 등이 알려져 있다. 본 연구에서는 phorbol-12-myristate-13-acetate (PMA)로 자극된 인간 섬유육종 HT-1080 세포에서 5가지 활성검색방법 즉 : gelatin zymography, MMPs ELISA, wound healing assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot을 이용하여 비쑥의 조추출물과 그 용매 분획물의 MMP-2와 MMP-9 효소 활성에 대한 저해 효과를 평가하였다. 비쑥 시료들을 메틸렌 클로라이드(MC)와 메탄올(MeOH)로 각각 2번 추출하여 합한 것을 조추출물로 사용하였으며 이 조추출물은 MMP-2와 MMP-9 효소활성에 대해 유의한 억제 효과를 나타내었다. 이 조추출물은 용매극성에 따라 다시 n-hexane, 85% aq.MeOH, n-butanol 및 물 분획층으로 분획되었으며 이렇게 얻어진 4개의 용매 분획물들중에 n-hexane과 85% aq.MeOH 분획이 gelatin zymography와 MMP ELISA assay에서 MMP-2와 MMP-9의 활성을 효과적으로 억제하였다. 또한 wound healing assay, RT-PCR 및 Western blot assay에서 H₂O 분획을 제외한 모든 용매 분획물들이 세포 이동, 그리고 MMP-2 및 MMP-9의 mRNA와 단백질 발현을 유의하게 억제하였다. The halophyte Artemisia scoparia is an edible medicinal plant, with insecticidal, anti-inflammatory, anti-cholesterol, antipyretic, and antibacterial effects. The aim of this study was to assess the inhibitory effect of crude extract and solvent-partitioned fractions obtained from A. scoparia on MMP-2 and MMP-9 activity in phorbol-12-myristate-13-acetate (PMA)-stimulated human fibrosarcoma HT-1080 cells using four different activity tests: gelatin zymography, MMP enzyme-linked immunosorbent assay (ELISA), wound healing assay, reverse transcription-polymerase chain reaction (RT-PCR), and Western blot assay. A. scoparia samples were extracted twice with methylene chloride (MC) and twice with methanol (MeOH). After the MC and MeOH crude extracts were combined, the combined crude extracts showed a significant inhibitory effect against MMP-2 and MMP-9 enzymes. They were then fractionated into n-hexane, 85% (v/v) aqueous methanol (85% (v/v) aq.MeOH), n-butanol, and water according to solvent polarity. Among the four solvent-partitioned fractions, n-hexane and 85% (v/v) aq. MeOH fractions significantly inhibited MMP-2 and MMP-9 activity and cell mobility. In addition, the n-hexane and 85% (v/v) aq.MeOH fractions effectively inhibited MMP-2 and -9 activity in the gelatin zymography and MMP ELISA assay. In the wound healing assay, RT-PCR, and Western blot assay, all solvent-partitioned fractions, except the H₂O fraction, significantly suppressed cell migration, as well as the expression levels of MMP-2 and -9 mRNA and proteins.
타겟 샘플링 검사를 통한 출하품질 향상에 관한 사례 연구
김준세 ( Junse Kim ),이창기 ( Changki Lee ),김경남 ( Kyungnam Kim ),김창우 ( Changwoo Kim ),송혜미 ( Hyemi Song ),안성수 ( Seoungsu Ahn ),오재원 ( Jaewon Oh ),조현상 ( Hyunsang Jo ),한상섭 ( Sangseop Han ) 한국품질경영학회 2021 품질경영학회지 Vol.49 No.3
Purpose: For improving outgoing quality, this study presents a novel sampling framework based on predictive analytics. Methods: The proposed framework is composed of three steps. The first step is the variable selection. The knowledge-based and data-driven approaches are employed to select important variables. The second step is the model learning. In this step, we consider the supervised classification methods, the anomaly detection methods, and the rule-based methods. The applying model is the third step. This step includes the all processes to be enabled on real-time prediction. Each prediction model classifies a product as a target sample or random sample. Thereafter intensive quality inspections are executed on the specified target samples. Results: The inspection data of three Samsung products (mobile, TV, refrigerator) are used to check functional defects in the product by utilizing the proposed method. The results demonstrate that using target sampling is more effective and efficient than random sampling. Conclusion: The results of this paper show that the proposed method can efficiently detect products that have the possibilities of user's defect in the lot. Additionally our study can guide practitioners on how to easily detect defective products using stratified sampling.