• Biomolecules & Therapeutics
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• 제20권2호
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• pp.165-170
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• 2012

Cell migration plays a role in many physiological and pathological processes. Reactive oxygen species (ROS) produced in mammalian cells influence intracellular signaling processes which in turn regulate various biological activities. Here, we investigated whether melanoma cell migration could be controlled by ROS production under normoxia condition. Cell migration was measured by wound healing assay after scratching confluent monolayer of B16F10 mouse melanoma cells. Cell migration was enhanced over 12 h after scratching cells. In addition, we found that ROS production was increased by scratching cells. ERK phosphorylation was also increased by scratching cells but it was decreased by the treatment with ROS scavengers, N-acetylcysteine (NAC). Tumor cell migration was inhibited by the treatment with PD98059, ERK inhibitor, NAC or DPI, well-known ROS scavengers. Tumor cell growth as judged by succinate dehydrogenase activity was inhibited by NAC treatment. When mice were intraperitoneally administered with NAC, the intracellular ROS production was reduced in peripheral blood mononuclear cells. In addition, B16F10 tumor growth was significantly inhibited by in vivo treatment with NAC. Collectively, these findings suggest that tumor cell migration and growth could be controlled by ROS production and its downstream signaling pathways, in vitro and in vivo.

• Journal of Microbiology and Biotechnology
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• 제29권7호
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• pp.1009-1013
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• 2019

Polymeric nanoparticles are widely used for drug delivery due to their biodegradability property. Among the wide array of polymers, chitosan has received growing interest among researchers. It was widely used as a vehicle in polymeric nanoparticles for drug targeting. This review explored the current research on the antimicrobial activity of chitosan nanoparticles (ChNP) and the impact on the clinical applications. The antimicrobial activities of ChNP were widely reported against bacteria, fungi, yeasts and algae, in both in vivo and in vitro studies. For pharmaceutical applications, ChNP were used as antimicrobial coating for promoting wound healing, preventing infections and combating the rise of infectious disease. Besides, ChNP also exhibited significant inhibitory activities on foodborne microorganisms, particularly on fruits and vegetables. It is noteworthy that ChNP can be also applied to deliver antimicrobial drugs, which further enhance the efficiency and stability of the antimicrobial agent. The present review addresses the potential antimicrobial applications of ChNP from these few aspects.

• Molecules and Cells
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• 제42권8호
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• pp.589-596
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• 2019

${\beta}Pix$ is a guanine nucleotide exchange factor for the Rho family small GTPases, Rac1 and Cdc42. It is known to regulate focal adhesion dynamics and cell migration. However, the in vivo role of ${\beta}Pix$ is currently not well understood. Here, we report the production and characterization of ${\beta}Pix$-KO mice. Loss of ${\beta}Pix$ results in embryonic lethality accompanied by abnormal developmental features, such as incomplete neural tube closure, impaired axial rotation, and failure of allantois-chorion fusion. We also generated ${\beta}Pix$-KO mouse embryonic fibroblasts (MEFs) to examine ${\beta}Pix$ function in mouse fibroblasts. ${\beta}Pix$-KO MEFs exhibit decreased Rac1 activity, and defects in cell spreading and platelet-derived growth factor (PDGF)-induced ruffle formation and chemotaxis. The average size of focal adhesions is increased in ${\beta}Pix$-KO MEFs. Interestingly, ${\beta}Pix$-KO MEFs showed increased motility in random migration and rapid wound healing with elevated levels of MLC2 phosphorylation. Taken together, our data demonstrate that ${\beta}Pix$ plays essential roles in early embryonic development, cell spreading, and cell migration in fibroblasts.

• BMB Reports
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• 제57권5호
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• pp.250-255
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• 2024

Due to their stem-like characteristics and immunosuppressive properties, Mesenchymal stem cells (MSCs) offer remarkable potential in regenerative medicine. Much effort has been devoted to enhancing the efficacy of MSC therapy by enhancing MSC migration. In this study, we identified deubiquitinase BRCA1-associated protein 1 (BAP1) as an inhibitor of MSC migration. Using deubiquitinase siRNA library screening based on an in vitro wound healing assay, we found that silencing BAP1 significantly augmented MSC migration. Conversely, BAP1 overexpression reduced the migration and invasion capabilities of MSCs. BAP1 depletion in MSCs upregulates ERK phosphorylation, thereby increasing the expression of the migration factor, osteopontin. Further examination revealed that BAP1 interacts with phosphorylated ERK1/2, deubiquitinating their ubiquitins, and thus attenuating the ERK signaling pathway. Overall, our study highlights the critical role of BAP1 in regulating MSC migration through its deubiquitinase activity, and suggests a novel approach to improve the therapeutic potential of MSCs in regenerative medicine.

• 대한임상검사과학회지
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• 제50권3호
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• pp.337-344
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• 2018

발생하는 간암 중 가장 주요한 형태인 간세포암은 강한 전이특성으로 인해 높은 재발율과 사망률을 보인다. Silymarin은 엉겅퀴에서 추출한 플라보노이드 성분으로 여러 암세포주에서 상피간엽전환(epithelial mesenchymal transition, EMT) 조절을 통해 항암효과를 보이는 것으로 보고되었다. 본 연구에서는 silymarin이 EMT의 조절을 통해 간세포암 세포주인 Huh7 cell의 침윤과 전이를 억제하는지를 분석하고자 하였다. Huh7 cell의 침윤과 전이 활성을 분석하기 위하여 wound healing assay와 in vitro invasion assay를 시행하였고 EMT 관련 유전자와 상위 신호전달물질의 발현 분석을 위해 Western blot assay를 실시하였다. 그 결과 silymarin은 농도 의존적으로 Huh7 cell의 침윤과 전이를 억제하였다. EMT 관련 유전자 중 세포 부착 단백질인 E-cadherin은 증가하였으나, 중간엽세포의 지표인 vimentin, 종양미세환경 조절에 관여하는 MMP-9의 발현은 억제되었고 이들의 활성에 관여하는 전사인자인 Snail과 nuclear factor $(NF)-{\kappa}B$ 또한 농도 의존적으로 활성이 감소하는 것을 확인할 수 있었다. 특히, 상위신호전달물질 중 silymarin은 phosphoinositide-3-kinase (PI3K)/Akt의 인산화 억제를 통해 EMT 관련 유전자들을 조절하는 것으로 나타났고 이것은 selective inhibitor인 LY294002의 처리 결과로 확인할 수 있었다. 결과적으로, silymarin은 PI3K/Akt 경로를 통해 EMT 관련 유전자의 발현을 조절함으로써 Huh7 cell의 침윤과 전이를 억제하는 것으로 생각된다. 이를 통해 silymarin이 간세포암의 전이 억제에 효과적인 항암물질의 후보가 될 수 있는 잠재력을 가진 후보물질이 될 수 있음을 보여주었다.

• 생명과학회지
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• 제21권6호
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• pp.907-911
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• 2011

인간형 재조합단백질 각질세포 성장인자를 안정적으로 생산하는 누에 배양 세포(Bm5-hKGF cell)을 만들었다. 이 세포에서 분비되어 배지에 포함된 양은 15-20 ng/ml 정도였다. Bm5-hKGF cell에 누에의 PDI를 함께 발현시키면 세포외 분비량이 2배 증가하였다. Wound healing migration assay 결과 누에세포에서 생산된 인간형 재조합단백질 각질세포 성장인자는 세포생장을 촉진하는 활성을 가지고 있었다. 본 실험의 결과는 누에배양세포를 사용하여 저비용으로 양질의 인간형 재조합단백질을 대량생산 할 수 있는 것을 기대한다.

• 생명과학회지
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• 제21권7호
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• pp.923-931
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• 2011

본 연구에서는 식물체에 널리 분포하는 indole-3-carbinol (I3C)에 의한 OVCAR-3 인체 난소암세포의 이동성 및 침윤성 억제 가능성과 이와 연관된 기전을 조사하였다. 본 연구의 결과에 의하면 I3C에 의한 OVCAR-3 세포의 증식억제는 세포의 이동성 억제와 연관이 있었으며, 이를 wound healing 및 matrigel invasion assay로 확인 하였다. 아울러 I3C 처리에 의하여 transepithelial electrical resistance가 증가되었으며, cellular paracellular permeability는 감소되었는데, 이는 I3C 처리에 의해 세포 내 치밀결합(tight junctions, TJs)의 tightness가 증가되었음을 의미한다. RT-PCR 및 immunoblotting 결과에 의하면, I3C는 TJs의 구성 성분이면서 paracellular transport의 선택적 투과성을 조절하는 주요 인자인 claudin-3 및 -4의 발현을 유의적으로 억제하였다. 또한 matrix metalloproteinase (MMP)-2 및 -9의 활성이 I3C 처리에 의하여 매우 억제되었는데, 이는 그들의 mRNA 및 단백질 수준에서의 발현 감소와 연관성이 있었다. 따라서 I3C에 의한 OVCAR-3 난소암세포의 침윤성 억제는 TJs 기능의 강화와 MMP 활성의 저하가 주요 인자로 작용함을 알 수 있었다.

• 생명과학회지
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• 제32권10호
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• pp.749-761
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• 2022

본 연구는 갯강활 추출물과 그 용매분획물의 항산화 및 암세포증식 억제효과를 평가하고자 하였다. 갯강활의 건조 시료를 차례대로 메틸렌클로라이드(CH₂Cl₂)로 2회, 그리고 메탄올(MeOH)로 2회 추출한 다음, 그 조추출물을 합한 후에 다시 용매극성에 따라 n-hexane, 85% 메탄올 수용액(85% aq.MeOH), n-buta- nol(n-BuOH) 및 물 분획층으로 분획하였다. 합해진 조추출물과 용매분획물의 항산화 활성은 DPPH 라디칼과 peroxynitrite 소거능, 세포내 활성산소종(ROS) 생성, DNA 산화, NO 생성, 철이온 환원력(FRAP)에 의해 평가되었다. 조추출물은 모든 항산화활성검색 시스템에서 유의적인 항산화 활성을 보였다. 용매분획들 중에서는 n-BuOH 및 85% aq.MeOH 분획에서 우수한 항산화 활성이 관찰되었으며 이 활성은 시료의 폴리페놀 및 플라보노이드 함량과 유의적인 상관관계가 있었다. 이 뿐만 아니라 조추출물을 포함한 모든 시료들이 인간 암세포(AGS, HT-29, MCF-7, HT-1080)에 대한 세포 독성 효과를 보였으며, HT 1080을 이용한 wound healing assay에서도 농도 의존적으로 세포이동을 억제하였다. 시료 중에는 85% aq.MeOH 용매분획이 HT-1080 세포의 침입을 가장 효과적으로 억제하였다. 그러므로 본 연구결과는 갯강활을 이용하여 항산화제 및 항암제 개발을 위한 기초자료로 활용될 수 있을 것이다.

• 대한한방내과학회지
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• 제37권1호
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• pp.16-25
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• 2016

Objectives: Gilgyung-tang (GGT) has been used as one of the main multi-herb formulas to treat “Peo-ong” (lung abscess). In this study, we investigated the inhibitory effects of water extracts of GGT on cell migration and invasion, two critical cellular processes that are often deregulated during metastasis, in human bladder cancer 5637 cells.Methods: Effects on cell viability were quantified using an MTT assay. To analyze the anti-metastatic effects, we conducted a wound healing migration assay, an in vitro invasiveness assay, and a measurement of the transepithelial electrical resistance (TER). The expression of protein and mRNA were measured by Western blotting and real-time polymerase chain reaction (RT-PCR), respectively.Results: GGT markedly inhibited the cell motility and invasiveness of 5637 cells within the concentration range that was not cytotoxic. The inhibitory effects of GGT on cell invasiveness were associated with tightening of the tight junctions (TJs), which was demonstrated by an increase in the TER. The RT-PCR and Western blotting results indicated that GGT decreased the levels of claudin proteins. GGT also inhibited the activity and expression of matrix metalloproteinase (MMP)-2 and -9 and simultaneously increased the levels of tissue inhibitor of metalloproteinase-1 and -2.Conclusions: Our findings suggest that GGT reduces both the migration and the invasion of 5637 cells by modulating the activity of TJs and MMPs.

• Asian Pacific Journal of Cancer Prevention
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• 제15권2호
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• pp.785-791
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• 2014

Crude extracts or phytochemicals obtained from some plants have potential anti-cancer properties. Teucrium persicum is an Iranian endemic plant belonging to the Lamiaceae family which has traditionally been used to relieve abdominal pains. However, the anti-cancer properties of this species of the Teucrium genus have not been investigated previously. In this study, we have used a highly invasive prostate cancer cell line, PC-3, which is an appropriate cell system to study anti-tumor properties of plants. A methanolic extract obtained from T persicum potently inhibited viability of PC-3 cells. The viability of SW480 colon and T47D breast cancer cells was also significantly decreased in the presence of the T persicum extract. Flow cytometry suggested that the reduction of cell viability was due to induction of apoptosis. In addition, the results of wound healing and gelatin zymography experiments supported anti-cell invasion activity of T persicum. Interestingly, sublethal concentrations of T persicum extract induced an epithelial-like morphology in a subpopulation of cells with an increase in E-Cadherin and ${\beta}$-Catenin protein levels at the cell membrane. These results strongly suggest that T persicum is a plant with very potent anti-tumor activity.