HT-1080 세포주에서 좀보리사초 추출물의 MMP-2와MMP-9 활성 억제효과

김준세,공창숙,서영완 한국해양과학기술원 2018 Ocean and Polar Research Vol.40 No.4

Matrix metalloproteinases (MMPs) are associated with the invasion and metastasis of malignant tumors composed of cancer cells in an increased state of expression. This study evaluates the inhibitory effect of Carex pumila on MMP-2 and MMP-9 activity in phorbol-12-myristate-13-acetate (PMA)- stimulated HT-1080 human fibrosarcoma cells using gelatin zymography, MMPs enzyme-linked immunosorbent assay (ELISA), reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assay. C. pumila was extracted twice with dichloromethane (CH2Cl2) and methanol (MeOH). Treatment with CH2Cl2 extract and MeOH extract in PMA-stimulated HT-1080 cells effectively reduced the production of MMP-2 and 9. Also, the combined crude extracts (CH2Cl2 and MeOH) significantly inhibited the enzymatic activities and the expression of MMP-2 and MMP-9 in mRNA and protein levels. The combined crude extracts were partitioned between CH2Cl2 and water. The organic layer was further fractionated with n-hexane, 85% aqueous methanol (85% aq.MeOH) and the aqueous layer was separated into n-butanol and water, successively. Of the fractions, 85% aq.MeOH fraction showed the highest inhibitory activity of MMP-2 and MMP-9 in gelatin zymography and MMP ELISA kit. Furthermore, 85% aq.MeOH fraction most significantly suppressed cell migration. In RT-PCR and Western blot assay, nbutanol and 85% aq.MeOH fractions exerted the greatest inhibition on mRNA and protein expression of MMP-2 and MMP-9, respectively. As a result, C. pumila can be used as a good anti-invasive agent source.

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.

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.

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.

가는갯능쟁이(Atriplex gmelinii) 추출물과 용매분획물의 MMP-2와 MMP-9 활성 저해효과

박민정,김준세,공창숙,서영완 한국해양과학기술원 2019 Ocean and Polar Research Vol.41 No.2

In this study, the inhibitory effect of Atriplex gmelinii C. A. Mey. against the activity of MMP-2 and MMP-9 secreted from phorbol-12-myristate-13-acetate (PMA)-stimulated HT-1080 cells was evaluated by gelatin zymography and enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase-chain reaction (RT-PCR), and Western blot assay. Specimens of the halophyte A. gmelinii were extracted twice for 24 hr with methylene chloride (CH2Cl2), and then twice with methanol (MeOH), in turn. Each extract significantly inhibited the enzymatic activities in gelatin zymography and MMP ELISA kit, and expression of MMP-2 and 9 in mRNA and protein levels. Two crude extracts were combined and then the combined crude extracts were fractionated into n-hexane, 85% aqueous methanol (85% aq.MeOH), nbutanol (n-BuOH), and water (H2O) fractions, according to solvent polarity. Among solvent-partitioned fractions, the 85% aq.MeOH fraction showed the strongest inhibitory effect against MMP-2 and -9 in gelatin zymography and MMP ELISA kit. In RT-PCR, all solvent-partitioned fractions significantly suppressed mRNA expression of MMP-2 and -9. On the other hand, in Western blot assay, all solventpartitioned fractions except H2O significantly reduced expression levels of protein. HT 1080 cell migration was most significantly inhibited by the n-BuOH fraction followed by the 85% aq.MeOH and H2O fractions. These results suggest that A. gmelinii could be used as a potential source to inhibit tumor cell metastasis.

Novel Discovery of LINE-1 in a Korean Individual by a Target Enrichment Method

신원석,문세영,김준세,이우석,박동국,최승규,이태윤,차승희,한규동 한국분자세포생물학회 2019 Molecules and cells Vol.42 No.1

Long interspersed element-1 (LINE-1 or L1) is an autonomous retrotransposon, which is capable of inserting into a newregion of genome. Previous studies have reported that these elements lead to genomic variations and altered functions byaffecting gene expression and genetic networks. Mounting evidence strongly indicates that genetic diseases or variouscancers can occur as a result of retrotransposition events that involve L1s. Therefore, the development of methodologies tostudy the structural variations and interpersonal insertion polymorphisms by L1 element-associated changes in an individual genome is invaluable. In this study, we applied a systematic approach to identify human-specific L1s (i.e., L1Hs)through the bioinformatics analysis of high-throughput nextgeneration sequencing data. We identified 525 candidatesthat could be inferred to carry non-reference L1Hs in a Korean individual genome (KPGP9). Among them, we randomlyselected 40 candidates and validated that approximately 92.5% of non-reference L1Hs were inserted into a KPGP9 genome. In addition, unlike conventional methods, our relatively simple and expedited approach was highly reproduciblein confirming the L1 insertions. Taken together, our findings strongly support that the identification of non-reference L1Hsby our novel target enrichment method demonstrates its future application to genomic variation studies on the risk of cancer and genetic disorders.

갯메꽃 추출물과 유기용매 분획물의 암전이 억제 효과

손재민(Jaemin Son),김준세(Junse Kim),김호준(Hojun Kim),서영완(Youngwan Seo) 한국생명과학회 2019 생명과학회지 Vol.29 No.3

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.

캠퍼스 내 감성적 정보전달을 위한 디지털 미디어 개발

우병현(Woo Byung-Hyun),김준세(Kim Jun-Se),김유진(Kim Yu-Jin),홍석기(Hong suk-ki) 한국디자인학회 2007 한국디자인학회 학술발표대회 논문집 Vol.- No.-