• Clinical and Experimental Reproductive Medicine
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• 제40권1호
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• pp.12-22
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• 2013

Objective: We investigated the norepinephrine transporter (NET) expression in normal and pre-eclamptic placentas and analyzed the invasion activity of trophoblastic cells based on norepinephrine (NE)-NET regulation. Methods: NET and NE expression levels were examined by western blot and enzyme-linked immunosorbent assay, respectively. Trophoblast invasion activity, depending on NE-NET regulation, was determined by NET-small interfering RNA (siRNA) and NET transfection into the human extravillous trophoblast cells with or without NE treatment and invasion rates were analyzed by zymography and an invasion assay. Results: NET mRNA was expressed at a low level in pre-eclamptic placentas compared with normal placentas and NE concentration in maternal plasma increased significantly in pre-eclamptic women compared to normal pregnant women (p<0.05). NET gene upregulation and NE treatment stimulated trophoblast cell invasion up to 2.5-fold (p<0.05) by stimulating matrix metalloproteinase-9 activity via the phosphoinositol-3-kinase/AKT signaling pathway, whereas NET-siRNA with NE treatment reduced invasion rates. Conclusion: NET expression is reduced by inadequate regulation of NE levels during placental development. This suggests that a complementary balance between NET and NE regulates trophoblast cell invasion activities during placental development.

• Journal of Korean Neurosurgical Society
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• 제46권5호
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• pp.472-478
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• 2009

Objective : Hyaluronidase (HAse), a degrading enzyme of hyaluronic acid (HA), is highly expressed in patients with malignant glioma. The purpose of this study was to verify whether HAse is related to the invasion of glioma cells. We also investigated if glioma cells with higher mobility in 2-dimensioal (2-D) method have also higher mobility at 3-dimensional (3-D) environment. Methods : Malignant glioma cell lines (U87MG, U251MG, U343MG-A, and U373MG) were used, and their HAse expressions were evaluated by HA zymography. The migration ability was evaluated by simple scratch technique. The invasiveness of each cell lines was evaluated by Matrigel invasion assay and HA hydrogel invasion assay. In HA hydrogel invasion assay, colonies larger than $150\;{\mu}m$ were regarded as positive ones and counted. Statistical analysis of migration ability and invasion properties of each cell lines was performed using t-test. Results : In scratch test to examine migration ability of each cell lines, U87MG cells were most motile than others, and U343MG-A least motile. The HAse was expressed in U251MG and U343MG-A cell lines. However, U87MG and U373MG cell lines did not express HAse activity. In Matrigel invasion assay, the cell lines expressing HAse (U251MG and U343MG-A) were more invasive in the presence of HA than HAse deficient cell lines (U87MG and U373MG). In HA hydrogel invasion assay, the HAse-expressing cell lines formed colonies more invasively than HAse-deficient ones. Conclusion : Malignant Glioma cells expressing HAse were more invasive than HAse-deficient ones in 3-dimensional environment. Therefore, it might be suggested that invasion of malignant gliomas is suppressed by inhibition of HAse expression or HA secretion. Additionally, the ability of 2-D migration and 3-D invasion might not be always coincident to each other in malignant glioma cells.

• 대한약침학회지
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• 제21권3호
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• pp.177-184
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• 2018

Objectives: Auraptene, a natural citrus coumarin, found in plants of Rutaceae and Apiaceae families. In this study, we investigated the effects of auraptene on tumor migration, invasion and matrix metalloproteinase (MMP)-2 and -9 enzymes activity. Methods: The effects of auraptene on the viability of A2780 and Hela cell lines was evaluated by MTT assay. Wound healing migration assay and Boyden chamber assay were determined the effect of auraptene on migration and cell invasion, respectively. MMP-2 and MMP-9 activities were analyzed by gelatin zymography assay. Results: Auraptene reduced A2780 cell viability. The results showed that auraptene inhibited in vitro migration and invasion of both cells. Furthermore, cell invasion ability suppressed at $100{\mu}M$ auraptene in Hela cells and at 25, $50{\mu}M$ in A2780 cell line. Gelatin zymography showed that for Hela cell line, auraptene suppressed MMP-2 enzymatic activity in all concentrations and for MMP-9 at a concentration between 12.5 to $100{\mu}M$ in A2780 cell line. Conclusion: Auraptene inhibited migration and invasion of human cervical and ovarian cancer cells in vitro by possibly inhibitory effects on MMP-2 and MMP-9 activity.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.327.3-328
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• 2002

Transforming growth factor-${\beta}$ (TGF-${\beta}$), a hormonally active polypeptide found in normal and transformed tissues. regulates cellular growth and phenotyphic plasticity. We have previously shown that H-ras. but not N-ras. induces invasive phenotype in MCF10A human breast epithelial cells. In this study. we wished to examine the effect of TGF-${\beta}$ on H-ras-induced invasion and motility in MCFI 10A cells by performing in vitro invasion assay and wound migration assay. (omitted)

• Asian Pacific Journal of Cancer Prevention
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• 제13권9호
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• pp.4815-4822
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• 2012

Invasion and metastasis are the major causes of cancer-related death. Pharmacological or therapeutic interventions such as chemoprevention of the progression stages of neoplastic development could result in substantial reduction in the incidence of cancer mortality. (-)-Epigallocatechin-3-gallate (EGCG), a promising chemopreventive agent, has attracted extensive interest for cancer therapy utilizing its antioxidant, anti-proliferative and inhibitory effects on angiogenesis and tumor cell invasion. In this study, we assessed the influence of EGCG on the proliferative potential of HeLa cells by cell viability assay and authenticated the results by nuclear morphological examination, DNA laddering assay and cell cycle analysis. Further we analyzed the anti-invasive properties of EGCG by wound migration assay and gene expression of MMP-9 and TIMP-1 in HeLa cells. Our results indicated that EGCG induced growth inhibition of HeLa cells in a dose- and time-dependent manner. It was observed that cell death mediated by EGCG was through apoptosis. Interestingly, EGCG effectively inhibited invasion and migration of HeLa cells and modulated the expression of related genes (MMP-9 and TIMP-1). These results indicate that EGCG may effectively suppress promotion and progression stages of cervical cancer development.

• 대한한방종양학회지
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• 제7권1호
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• pp.109-116
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• 2001

By applying in vitro invasion assay model, we examined the anti-metatstastic effect of ChunghjagamTang(CLJGT). In 3H-thymidine incorporation assay, CLJGT treated groups showed the decreased DNA synthesis rate compared with control group. Gelatin zymogram assay showed that CLJGT decreases the gelatinolytic activity of MMP-9 from A-549, at the concentration of $800{\mu}g/ml$. We examined whether CLJGT inhibits the invasion of A-549 cells through the matrigel precoated transwell chamber. The results showed that CLJGT effectively inhibited the invasion of A-549 as compared with the control (+PMA) groups. From our research, part of mechanism underlying anti-metastastic effect of CLJGT was proven in vitro.

• 환경생물
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• 제28권1호
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• pp.8-13
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• 2010

본 연구는 BRECs에서 AGEs로 유도된 세포의 이동 및 침윤에 있어서 AuNP의 역할에 관한 연구이다. 소 망막으로부터 내피세포를 분리하고, 세포 생존율은 MTT assay로 확인하였다. Wound migration assay는 BRECs의 이동력을 확인하기 위해 수행하였다. Tube formation은 on-gel system을 통해 확인하였다. AuNP의 apoptosis 유도는 caspase-3 assay를 통해 확인하였다. AGE-BSA은 세포증식 및 이동에 있어서 증가함을 보여주었다. 또한 AuNP는 AGE-BSA 존재 유무에 상관없이 세포의 증식, 이동, 신생혈관 형성을 억제하였고, caspase-3을 통해 apoptosis를 유도하였다. 이러한 결과, AuNP는 AGE로 유도된 신생혈관 형성 및 세포의 이동성을 억제하는 약재제로서, 당뇨성 합병증에 있어서 잠재적으로 중요한 분자가 될 것이다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.5533-5537
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• 2013

MicroRNA-10b (miR-10b) has been reported to play an important role in some types of cancer, but the effects and possible mechanisms of action of miR-10b in the metastasis of nasopharyngeal carcinoma cells (NPC) have not been explored. The aim of the present study was to investigate the function of miR-10b in nasopharyngeal carcinoma and to determine the molecular mechanisms underlying its action. The MTT assay was used to assess proliferation of CNE-2Z cells. Wound healing and transwell migration assays were applied to assess cell migration and invasion, while and expression of E-cadherin and MMP-9 were detected using Western blot analysis. Real-time PCR was employed to detect the expression of genes related to migration and invasion and the $2^{-{\Delta}{\Delta}Ct}$ method was used to calculate the degree of expression. MTT assay showed the expression of miR-10b to have no effect on the proliferation of NPC cell lines. The wound healing assay showed that miR-10b mimics promoted the mobility and invasion of NPC cell lines. Inhibitors of miR-10b reduced the ability of NPC cell lines to migrate and invade. In addition, the expression of genes related to migration and invasion, such as E-cadherin, vimentin, and MMP-9, were confirmed to be different in the CNE-2Z NPC cell line transfected with miR-10b mimics and with miR-10b inhibitors. In the present study, miR-10b was found to upregulate the expression of MMP-9 and knockdown of miR-10b was found to significantly downregulate the expression of E-cadherin. On the whole, these results showed that miR-10b plays an important role in the invasion and metastasis of NPC cells.

• 대한한방내과학회지
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• 제35권4호
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• pp.416-427
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• 2014

Objectives: The purpose of this study was to investigate the antineoplastic effect of several herbal medicine mixtures (compositions of Astragalus membranaceu, Angelica gigas, Trichosanthes kirilowii, Panax ginseng, Rhus verniciflua Stokes) on the SNU-80 anaplastic thyroid carcinoma cell line. Methods: MTT assay was used to examine whether our herbal medicine mixtures decreased cell growth rate of SNU-80. Wound healing assay and Transwell invasion assay was performed to investigate whether our herbal medicine mixtures affect the migration and invasion of anaplastic cancer cells, SNU-80. ELISA assay was performed to know if our herbal medicine mixtures suppressed the expression of pro-invasive molecules, such as vascular endothelial growth factor (VEGF) and matrix metalloproteinase-2 (MMP-2) secreted from SNU-80. Results: MTT assay demonstrated that A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas :T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 strongly suppressed the growth of SNU-80. Wound healing assay demonstrated that A. membranaceus:A. gigas=3:1, A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 inhibited the migration of SNU-80. Transwell invasion assay demonstrated that A. membranaceus:A. gigas=1:1, A. membranaceus:A. gigas:T. kirilowii =1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng=1:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng :R. verniciflua Stokes=1:1:1:1:1 or 3:1:1:1:1 inhibited the invasion of SNU-80. ELISA assay demonstrated that A. membranaceus :A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes=1:1:1:1:1 suppressed the expression of VEGF. Also, A. membranaceus:A. gigas=1:1, A. membranaceus:A. gigas:T. kirilowii=1:1:1 or 3:1:1, A. membranaceus :A. gigas:T. kirilowii:P. ginseng=1:1:1:1 or 3:1:1:1, A. membranaceus:A. gigas:T. kirilowii:P. ginseng:R. verniciflua Stokes =1:1:1:1:1 or 3:1:1:1:1 suppressed the expression of MMP-2. Conclusions: The results obtained in this study suggest that several herbal medicine mixtures suppresse the growth and inhibit the migration and invasion of SNU-80, which is anaplastic thyroid cancer cells. Especially, A. membranaceus:A. gigas: T. kirilowii=1:1:1 mixture had a stronger anti-cancer effect.

• 대한예방한의학회지
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• 제18권1호
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• pp.83-92
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• 2014

Objective : This study was performed to investigate the antineoplastic effect of Astragalus membranaceus, Angelica gigas, Rhus verniciflua Stokes and Trichosanthes kirilowii on SNU-80 anaplastic thyroid carcinoma cell line. Method : We examined whether our herbal medicines decreases cell growth rate of SNU-80 using MTT assay. We performed western blot analysis to verify that our herbal medicines induces apoptosis via caspase-dependent mechanism. We also performed wound healing assay and transwell invasion assay to investigate whether our herbal medicines affects the migration and invasion of anaplastic cancer cells, SNU-80. We also carried out ELISA assay to know our herbal medicines suppresses the expression of proinvasive molecules, such as VEGF and MMP-2 secreted from SNU-80. Results : MTT assay demonstrates that Angelica gigas, Rhus verniciflua Stokes, and Trichosanthes kirilowii suppressed strongly the growth of SNU-80. Western blot analysis demonstrates that Trichosanthes kirilowii induces apoptosis activating the cleavages of caspases (caspase-8, caspase-3) and PARP. Wound healing assay demonstrates that Rhus verniciflua Stokes, and Trichosanthes kirilowii inhibited the migration of SNU-80. Transwell invasion assay demonstrates that Rhus verniciflua Stokes, and Trichosanthes kirilowii inhibited the invasion of SNU-80. Elisa assay demonstrates that Astragalus membranaceus, Angelica gigas, Rhus verniciflua Stokes, and Trichosanthes kirilowii suppressed the expression of VEGF and MMP-2. Conclusion : We could conclude that several herbal medicines suppresses the growth and inhibits the migration and invasion of SNU-80 which is anaplastic thyroid cancer cells. Especially, Rhus verniciflua Stokes, Trichosanthes kirilowii had stronger anti-cancer effect suggesting that we can apply them to treat anaplastic thyroid cancer.