• 한국식품과학회지
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• 제50권1호
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• pp.105-110
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• 2018

멜리틴은 봉독의 주요 성분 중 하나로 항염증과 항암활성 효과를 가지고 있다. 우리는 폐암세포에서 멜리틴이 EMT 억제를 통해 암세포 이동과 침투를 억제하는 사실을 확인하였다. 멜리틴은 EGF로 유도된 폐암 세포 이동과 침투를 억제하였을 뿐만 아니라 EMT와 관련된 단백질인 이카드헤린의 발현을 증가시켰으며, 바이멘틴과 피브로넥틴 발현은 감소시켰다. 또한 멜리틴에 의한 EMT조절 전사인자인 ZEB2, Slug, Snail의 발현을 확인한 결과 멜리틴 처리에 의해 농도의존적으로 발현이 감소하였다. 또한 작용 메커니즘을 확인하기 위해 mTOR와 FAK 메커니즘을 확인한 실험에서 EGF 처리에 의해 증가한 AKT, mTOR, p70S6K, 4EBP1의 인산화가 멜리틴 농도의존적으로 감소하였다. 그러나 FAK는 EGF에 의해 변화가 없었으며, EKR, JNK 메커니즘은 EGF 처리에 의해 인산화가 증가하였으나 멜리틴 처리에 의해 아무런 영향을 받지 않았다. 그러므로, 폐암세포의 세포 이동과 침투에 대한 멜리틴의 억제효과는 AKT/mTOR/P70S6K/4EBP1 기전 억제를 통해 EMT를 억제하여 세포 이동과 침투를 억제하는 것으로 보인다.

• Journal of Ginseng Research
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• 제45권1호
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• pp.134-148
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• 2021

Background: Lung cancer has a high incidence worldwide, and most lung cancer-associated deaths are attributable to cancer metastasis. Although several medicinal properties of Panax ginseng Meyer have been reported, the effect of ginsenosides Rk1 and Rg5 on epithelial-mesenchymal transition (EMT) stimulated by transforming growth factor beta 1 (TGF-β1) and self-renewal in A549 cells is relatively unknown. Methods: We treated TGF-β1 or alternatively Rk1 and Rg5 in A549 cells. We used western blot analysis, real-time polymerase chain reaction (qPCR), wound healing assay, Matrigel invasion assay, and anoikis assays to determine the effect of Rk1 and Rg5 on TGF-mediated EMT in lung cancer cell. In addition, we performed tumorsphere formation assays and real-time PCR to evaluate the stem-like properties. Results: EMT is induced by TGF-β1 in A549 cells causing the development of cancer stem-like features. Expression of E-cadherin, an epithelial marker, decreased and an increase in vimentin expression was noted. Cell mobility, invasiveness, and anoikis resistance were enhanced with TGF-β1 treatment. In addition, the expression of stem cell markers, CD44, and CD133, was also increased. Treatment with Rk1 and Rg5 suppressed EMT by TGF-β1 and the development of stemness in a dose-dependent manner. Additionally, Rk1 and Rg5 markedly suppressed TGF-β1-induced metalloproteinase-2/9 (MMP2/9) activity, and activation of Smad2/3 and nuclear factor kappa B/extra-cellular signal regulated kinases (NF-kB/ERK) pathways in lung cancer cells. Conclusions: Rk1 and Rg5 regulate the EMT inducing TGF-β1 by suppressing the Smad and NF-κB/ERK pathways (non-Smad pathway).

• The Korean Journal of Physiology and Pharmacology
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• 제26권4호
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• pp.239-253
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• 2022

Epithelial-mesenchymal transition (EMT) is known to be involved in airway remodeling and fibrosis of bronchial asthma. However, the molecular mechanisms leading to EMT have yet to be fully clarified. The current study was designed to reveal the potential mechanism of microRNA-21 (miR-21) and poly (ADP-ribose) polymerase-1 (PARP-1) affecting EMT through the PI3K/AKT signaling pathway. Human bronchial epithelial cells (16HBE cells) were transfected with miR-21 mimics/inhibitors and PARP-1 plasmid/small interfering RNA (siRNA). A dual luciferase reporter assay and biotin-labeled RNA pull-down experiments were conducted to verify the targeting relationship between miR-21 mimics and PARP-1. The migration ability of 16HBE cells was evaluated by Transwell assay. Quantitative real-time polymerase chain reaction and Western blotting experiments were applied to determine the expression of Snail, ZEB1, E-cadherin, N-cadherin, Vimentin, and PARP-1. The effects of the PI3K inhibitor LY294002 on the migration of 16HBE cells and EMT were investigated. Overexpression of miR-21 mimics induced migration and EMT of 16HBE cells, which was significantly inhibited by overexpression of PARP-1. Our findings showed that PARP-1 was a direct target of miR-21, and that miR-21 targeted PARP-1 to promote migration and EMT of 16HBE cells through the PI3K/AKT signaling pathway. Using LY294002 to block PI3K/AKT signaling pathway resulted in a significant reduction in the migration and EMT of 16HBE cells. These results suggest that miR-21 promotes EMT and migration of HBE cells by targeting PARP-1. Additionally, the PI3K/AKT signaling pathway might be involved in this mechanism, which could indicate its usefulness as a therapeutic target for asthma.

• 한국발생생물학회지:발생과생식
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• 제12권1호
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• pp.87-95
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• 2008

만능성 인간 배아줄기세포로부터 확립된 신경줄기세포 또는 신경전구세포는 퇴행성 신경질환 세포치료제로 이용될 수 있는 다양한 종류의 신경세포로 분화 유도될 수 있다. 하지만, 인간 배아줄기세포로부터 신경세포를 생산하기 위한 기술은 아직 많은 장애를 가지고 있다. 인간 배아줄기세포 유래 신경전구세포에서 특징적으로 나타나는 신경관 유사로제트에 대한 이해는 인간 배아줄기세포 신경 분화의 효율을 높이는데 유용한 정보를 제공할 것으로 사료된다. 일반적으로 신경로제트(neural rosette)는 분화 중인 배아체를 부착 배양함으로써 유도하지만, 이 방법은 시간이 걸리고 복잡하다는 단점이 있다. 본 연구에서는 신경로제트가 부착배양을 하지 않고 부유배양으로 형성될 수 있는지 조사하였다. 우선적으로, 배아체 형성 및 신경분화에 인간 배아줄기세포 클럼프(clump) 크기가 영향을 주는지를 조사하였고, 사방 $500\;{\mu}m$ 크기의 인간 배아줄기세포 클럼프가 신경 분화 유도에 가장 효과적임을 확인하였다. 로제트 형성을 유도하기 위해, 사방 $500\;{\mu}m$ 크기의 인간 배아줄기세포 클럼프를 1주일 동안 EB 배양배지에 부유 배양함으로써 균일한 크기의 배아체를 얻은 후, NES 배양 배지에서 부가적으로 $1{\sim}2$주 동안 계속 부유 배양한 결과, $7{\sim}10$일 사이에 신경관 유사 로제트가 형성됨을 확인하였다. 로제트 형성 세포의 신경전구세포로서 특성은 RT-PCR과 면역형광염색법을 이용한 신경전구세포 특이적 마커(vimentivi, nestin, MSI1, MSI2, Sox1, Tuj1) 발현을 통해 확인하였다. 또한, 성장인자를 제외한 NES 배양 배지에서 신경로제트를 $2{\sim}6$주 동안 지속적으로 배양하면 성숙 신경세포로의 말단 분화가 유도됨을 확인하였다. 신경세포 특이적 마커(Tuj1, MAP2, GABA)와 신경아교 특이적 마커($S100{\beta}$, GFAP)는 $2{\sim}3$주 또는 4주 후에 각각 발현이 유도됨을 확인하였고, 희소 돌기아교 특이적 마커(O1과 CNPase)는 $5{\sim}6$주 후에 발현이 증가함을 확인하였다. 본 연구결과는 신경로제트가 부유 배양시스템에서 성공적으로 형성됨을 보여주고 있으며, 이는 인간 배아줄기세포의 신경 분화를 이해하고, 신경전구세포 유도 과정을 단순화하는데 효과적으로 이용될 수 있을 것으로 사료된다.

• 대한물리치료과학회지
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• 제9권3호
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• pp.107-119
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• 2002

Astrocyte, which shares the greatest part of the brain (about 25%), is a land of glial cell that composes the central nervous system along with microglia, ependymal cell and oligodendroglia. It has 7-9nm of fibers in its cytoplasma, which are composed of glial fibrillary acidic protein (GFAP) and vimentin. As for the functions of the astrocyte, it has, so far, been supposed that the astrocyte will play a cytoskeletal role in maintaining the structure of the cerebrum, play a role as a blood-brain barrier so that it can induce migration of the neuron in its development and substances in the blood cannot go into the nervous tissue, and a role of immunology and phagocytosis. However, it was revealed today that it will be a role in preventing expansion of injury by attaching itself to the connective tissue such as the vessel and the pia mater when the nervous tissue or the arachnoid is injured. Microcurrent stimulation can control current, on the basis of A unit. That is, with such devices using it, it is possible to sense, from the outside, the injured current(wound current) of the lesion and to change it into the normal current, thereby promoting the restoration of the cells. In order to examine the effects of microcurrent stimulation on the injured astrocytes in the rabbits, this study was conducted with 24 New Zealand White Rabbit as its subjects, which were divided into 8 animals of the experiment group and 16 animals of the control group. After the animals in the experiment group were fixed to the stereotaxic apparatus, their hair was removed and their premotor area(association area) perforated by the micro-drill for skull-perforation with the depth of 8mm from the scalp. In one week after the injury, 4 animals in the control group and 8 animals in the experiment group were sacrificed and examined with immunohistochemical method. And in three weeks, the remaining 4 animals in the control group and 8 animals in the experiment group were also sacrificed and examined with the same way. The conclusion has been drawn as follows : In the control group sacrificed in one week after the injury, the astrocytes somewhat increased, compared with the normal animals, and in the group sacrificed in three weeks after the injury, they increased more (p < 0.05). The experiment group A in one week showed a little increase, but there was no significant differences, but the experiment group in three weeks showed more increase, compared with the experiment group in one week (p < 0.05). The experiment group B in one week showed more increase than the control group or the experiment group A, and the experiment group in three weeks showed more increase than the experiment group in one week (p < 0.05). Among the astrocytes, fibrous astrocytes were mostly observed, increasing as they are close to the lesion, and decreasing as they are remote from it. The findings show that microcurrent can cause the astrocytes to proliferate and that it will be more effective to stimulate the cervical part somewhat remote from the lesion rather than to directly stimulate the part of the lesion. Thus, microcurrent stimulation can be one of the methods that can activate the reaction of astrocytes, which is one of the mechanism for treating cerebral injury with hemorrhage. Therefore, this study will be used as basic research data for promoting restoration of functions in the patient with injury in the central nervous system.

• Asian Pacific Journal of Cancer Prevention
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• 제16권7호
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• pp.2659-2664
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• 2015

Background: To examine the expression of cysteine-rich 61 (Cyr61/CCN1) protein in laryngeal squamouscell carcinoma (LSCC) tissues, and its relationship with the tumor epithelial-mesenchymal transition (EMT), invasion, metastasis, and prognosis. Materials and Methods: Immunohistochemistry was used to detect the expressions of Cyr61, Vimentin (Vim), and E-cadherin (E-cad) in 88 cases of LSCC tissues and 30 cases of tumor-adjacent normal tissues. Vim and E-cad were used as mesenchymal and epithelial markers, respectively, to determine the relationship between Cyr61 expression and the EMT of LSCC cells. In addition, clinical and histopathological data were combined to analyze the relationship between the positive-expression rates of Cyr61, Vim and E-cad and LSCC invasion, metastasis and prognosis. Results: In LSCC tissues, Vim expression rate was significantly higher than that of the tumor-adjacent tissues, whereas E-cad expression rate was significantly lower than that of the tumor-adjacent tissues. The Vim expression rate was significantly higher in stages T3 and T4 than in stages T1 and T2 LSCC tissues, whereas E-cad expression rate was significantly lower in stages T3 and T4 than in stages T1 and T2 LSCC tissues. Compared to the group without lymph node metastasis, the Vim expression rate was significantly higher and the E-cad expression rate was significantly lower in the group with lymph node metastasis. The expression rate of Cyr61 was significantly higher in LSCC tissues than in the tumor-adjacent normal tissues. In addition, the Cyr61 expression rate was higher in stages T3 and T4 than in stages T1 and T2 LSCC, and higher in the group with lymph node metastasis than in the group without lymph node metastasis. The Vim expression rate was significantly higher in the Cyr61 positive group than in the Cyr61 negative group, whereas the E-cad expression rate was significantly higher in the Cyr61 negative group than in the Cyr61 positive group. Survival analysis indicated that survival rates of Cyr61 positive, Vim positive and E-cad negative groups were significantly lower than that of Cyr61 negative, Vim negative and E-cad positive groups, respectively. Conclusions: Cyr61 expression is closely associated with LSCC invasion and lymph node metastasis. Overexpression of Cyr61 may induce EMT and therefore leads to LSCC invasion and metastasis and poor prognosis. Cyr61 may become a new maker for clinical prediction of LSCC invasion and metastasis and a new target for LSCC treatment.