• Journal of Microbiology and Biotechnology
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• 제16권10호
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• pp.1544-1553
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• 2006

The halophilic enterobacteria, Enterobacteria cancerogenus, was isolated from the intestines of the fusiform fish (Trachurus japonicus) to yield a protein-like material termed PLM-f74. PLM-f74 was characterized by strong inhibition ratios to angiogenesis (82.8% at the concentration of $18.5{\mu}g/ml$) and elevated antioxidative capacities with low toxicity. The PLM-f74 is a glycoprotein comprised of saccharides and amino acids. PLM-f74 inhibited cell adhesion that non-activated U937 monocytic cell adhesion to HUVECs activated with $IL-1{\beta}$ by 78.0%, and the adherence of U937 cells treated with the PLM-f74 and stimulated with $IL-1{\beta}$ to unstimulated HUVECs decreased by 102%. When both cell types were pretreated with PLM-f74, the adhesion of U937 cells to $IL-1{\beta}$-stimulated HUVECs was completely suppressed by 121% at a concentration of $18.5{\mu}g/ml$. PLM-f74 blocked signal pathways from VEGFR2, PI3K, ${\beta}$-catenin, and VE-cadherin to NF-kB, based on western bolt analysis. It also inhibited IL-l-stimulated HUVEC expression of the adhesion molecules, ICAM-l by 40%, VCAM-l by 60%, and E-selectin by 70% at the same concentration noted above. New anti-angiogenic and anti-cell adhesion materials showing elevated antioxidative capacities, and non-toxicity may be expected from these results.

• Asian Pacific Journal of Cancer Prevention
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• 제15권4호
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• pp.1811-1815
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• 2014

Background: In clinical trials with no upper age limit, the proportion of older patients is usually small, probably reflecting the more conservative approach adopted by clinicians when treating the elderly. An exploratory analysis of elderly patients in the RECORD-1 Trial showed that patients ${\geq}$ 65 y.o. had superior median PFS than overall RECORD-1 population (5.4 months and 4.9 months, respectively). We investigated the efficacy, relative benefit and safety of Everolimus (EVE) as sequential therapy after failure of VEGFr-TKI therapy for older patients with metastatic renal cell cancer (mRCC), in daily practice. Materials and Methods: 172 consecutive IRB approved patients with mRCC (median age 65, M:F 135/37, 78% clear cell) who received salvage EVE at 39 tertiary institutions between October 2009 and August 2011 were included in this analysis. Some 31% had progressed on sunitinib, 22% on sorafenib, 1% on axitinib, 41% on sequential therapy, and 5% had received other therapy. Patients with brain metastases were not included and 95% of the patients had a ECOG (Eastern Cooperative Oncology Group) performance status (PS) of 0 or 1. Previous radiotherapy was an exclusion criterion, but prior chemotherapy was permitted. Adequate organ function and hematologic parameters were mandatory. EVE administration was approved by the institutional review board at each participating institution and signed informed consent was obtained from all patients. Results: Median time of the whole cohort to last follow-up was 3.5 months (range 0.4-15.2 months). Forty four percent were continuing to take EVE at last followup. There were 86 (50%) patients ${\geq}$ 65 y.o. and 86 (50%) <65 y.o. The percentage of patients who showed PR/SD was higher in the older group than in the younger one (5.9%/61.2% vs 1.2%/46.5%, respectively). Median survival of older patients was also significantly longer (3.5 +/- 0.31 vs 3.1 +/- 0.34, hazard ratio=0.45, CI; 0.255-0.802). Analysis using Cox regression model adjusted for gender, PS, number of metastases, site of metastases, histology, smoking history and age detected an association between age and PFS (p=0.011). The frequency of adverse events in elderly patients treated with EVE was no greater than that in younger patients, although such toxicity may have had a greater impact on their quality of life. Conclusions: Older patients should not generally be excluded from accepted therapies (mTOR inhibitors after failure of VEGFr-TKI therapy) for mRCC.

• The Korean Journal of Physiology and Pharmacology
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• 제22권6호
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• pp.627-636
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• 2018

Endometriosis (EM) is one of the most common gynaecological disorder affecting women in their reproductive age. Mechanisms involved in the pathogenesis of EM remains poorly understood, however inflammatory responses have been reported to be significantly involved. The efficacy of 6-shogaol on proliferation of endometriotic lesions and inflammatory pathways in experimentally-induced EM model was explored in this study. EM was stimulated in Sprague-Dawley rats by implantation of autologous endometrium onto the peritoneum abdominal wall. Separate groups were treated with 6-shogaol (50, 100 or 150 mg/kg b.wt/day) via oral gavage for one month period. Gestrinone (GTN) group received GTN (0.5 mg/kg/day) as positive control. Five weeks after implantation, the spherical volume of ecto-uterine tissues was determined. Treatment with 6-shogaol significantly reduced the implant size. Histological analysis reported atrophy and regression of the lesions. 6-shogaol administration effectively down-regulated $NF-{\kappa}B$ signaling, VEGF and VEGFR-2 (Flk-1) expression in the endometriotic lesions. Excess production of $IL-1{\beta}$ and IL-6 (pro-inflammatory cytokines), PGE2 and nitric oxide (NO) were reduced. Overall, the results of the study reveal the efficacy of 6-shogaol against endometriosis via effectively suppressing proliferation of the lesions and modulating angiogenesis and $COX-2/NF-{\kappa}B$-mediated inflammatory cascades.

• 대한임상검사과학회지
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• 제51권3호
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• pp.360-369
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• 2019

Cadherin은 원형질막에 존재하며 세포-세포 결합에 관여하며, 황체 구조 유지에 필수적인 단백질이다. 본 연구에서는 prostaglandin F2 alpha ($PGF2{\alpha}$)가 황체의 협막세포(luteal theca cells, LTCs)의 E-cadherin, N-cadherin 및 세포-세포부착에 미치는 영향에 대해서 수행하였다. 황체세포는 소의 황체중기 조직으로부터 분리하였으며, 황체세포 중에서 mesenchymal 세포 형태학적 특성을 가지는 세포만을 분리하여 LTCs로 판단하였다. 이 후 steroidogenic 기능 및 혈관세포 유무를 판단하기 위해 $3{\beta}$-HSD 및 VEGF2R mRNA 발현을 확인하였으며, E-cadherin 및 N-cadherin mRNA를 사용하여 LTCs 내 cadherin의 존재여부를 판단하였다. 또한 0, $10^{-5}$, $10^{-4}$ 및 $10^{-3}M$ $PGF2{\alpha}$를 24시간 동안 처리하여 LTCs의 E- 및 N-cadherin 단백질을 관찰한 후 세포-세포 접착 실험을 실시하였다. 그 결과, LTCs에서 $3{\beta}$-HSD mRNA가 발현되었지만, VEGFR2 mRNA는 발현되지 않았으며, E-cadherin 및 N-cadherin mRNA 모두 발현되는 것을 확인하였다. 또한 E-및 N-cadherin 단백질은 $10^{-5}$, $10^{-4}$ 및 $10^{-3}M$ $PGF2{\alpha}$를 처리한 LTCs에서 응집되어 발현되는 것을 확인하였으며, $PGF2{\alpha}$에 의해 LTCs의 세포부착 효율이 유의적으로 감소된 것을 확인하였다. 결론적으로 $PGF2{\alpha}$는 LTCs의 E- 및 N-cadherin을 붕괴시켜 세포부착을 감소시켰고, 이러한 결과는 황체퇴행의 새로운 원인을 밝혀 내기 위한 cadherin과 세포부착의 역할을 이해하는데 중요한 자료로 활용될 것으로 판단된다.

• 동의생리병리학회지
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• 제28권1호
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• pp.16-21
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• 2014

Micro needle roller therapy has been used for cosmetic purposes, such as reducing skin winkles and improving elasticity of skin. It is claimed that micro needle roller therapy has potentials for connective tissue regeneration by facilitating collagen synthesis. Therefore, there seems to be a possibility that connective tissue regenerating potential of micro needle roller therapy could influence the hair growth cycle. This study, we investigated the hair growth-promoting effects of micro needle roller therapy. C57BL/6 mice were devided into three groups as follows: normal saline-treated, minoxidil-treated, and micro needle roller therapy-received group. Hair growth activity was evaluated by handscopic and microscopic observations. Sections of dorsal skin were stained with hematoxylin and eosin. Expression of BrdU, FGF, and VEGF was detected by immunohistochemical staining. Micro needle roller therapy enhanced the development of hair follicle during anagen. Immunohistochemical analysis revealed that micro neeld roller therapy incresed the expression of BrdU and FGF in the hair follicles of C57BL/6 mice. Furthermore, micro needle roller therapy upregulated mRNA expression of VEGFR-2, FGF-2, EGF - growth factors that play a central role in hair follicle development during anagen. These results suggest that Micro needle roller therapy can potentially be used for the treatment of alopecia.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권1호
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• pp.59-70
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• 2008

Purpose: We evaluated the therapeutic effect of AEE788, a dual inhibitor of epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) receptor tyrosine kinases on human salivary adenoid cystic carcinoma (ACC) cells growing in nude mice. Experimental Design: We examined the effects of AEE788 on salivary ACC cell growth and apoptosis. To determine the in vivo effects of AEE788, nude mice with orthotopic parotid tumors were randomized to receive oral AEE788 (50 mg/kg) three times per week, injected paclitaxel ($200{\mu}g$) once per week, AEE788 plus paclitaxel, or placebo. Mechanisms of in vivo AEE788 activity were determined by immunohistochemical analysis. Results: Treatment of salivary ACC cells with AEE788 led to growth inhibition and induction of apoptosis. AEE788 inhibited tumor growth and prevented lung metastasis in nude mice. Furthermore, AEE788 potentiated growth inhibition and apoptosis of ACC tumor cells mediated by paclitaxel. Tumors of mice treated with AEE788 and AEE788 plus paclitaxel exhibited down-regulation of activated EGFR and its downstream mediators (Akt and MAPK), increased tumor and endothelial cell apoptosis, and decreased microvessel den-sity, which correlated with a decrease in the level of MMP-9, MMP-2 and bFGF expression and a decrease in the incidence of vascular metastasis. Conclusions: These data show that tumor-associated endothelial cells are important in the process of tumor-metastasis. And VEGFR can be a molecular target for therapy of metastatic lung lesion of salivary ACC.

• 생명과학회지
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• 제17권8호통권88호
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• pp.1152-1156
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• 2007

본 실험은 동백꽃의 효능을 알아보기 위하여, 동백꽃을 에틸알콜로써 추출한 후 극성이 다른 다양한 유기용매로 분획하여 나온 fraction으로 신생혈관생성 억제, cell cytotoxity, 세포부착분자에 대한 기능을 ELISA, westen blot 등으로 조사하였다. 신생혈관생성 억제 효과는 특히butanol 층에서 70.2%로 가장 높았으며, acetone층이 54.2%, 그리고 ethylacetate층이 37.0%, chloroform층이 21.2%로 유의성 있는 억제효과가 있었다. 특히 동백꽃의 butanol층은 신생혈관생성억제제로서 좋은 효과가 있는 것으로 판단된다. 그리고, 추출물의 농도별로 cell cytotoxity를 조사한 결과 농도가 200 ug/ml 이하에서 세포독성은 나타나지 않았다. Cell adhesion 저해율은 추출물의 농도 200 μg/mL 에서 ICAM-1은 52.9%, VCAM-1은 62.5%, 그리고 E-selectin은 35.7%로 나타났으며 동백꽃의 추출물의 성분 증가에 따라 발현이 감소됨으로써 cell adhesion을 저해하는 것으로 확인되었다. Westen blot 에서 첨가한 동백꽃 추출물의 농도가 높아질수록 신호전달분자의 발현이 약해지는 것을 관찰할 수 있었다. 따라서 신호전달 분자는 동백꽃 추출물에 의해서 신호전달이 차단되며, NF-KB를 억제함으로서 신생혈관생성을 저해하는 것으로 확인되었다. 따라서 동백꽃 추출물은 항암제 및 항비만제제로서 유용할 것으로 판단된다.

• 대한두경부종양학회지
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• 제27권1호
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• pp.32-37
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• 2011

서 론 : 미세갑상선유두상암종 환자에서 불필요한 예방적 중앙 림프절절제술을 피하기 위해서 림프절 전이를 수술 전에 예측하는 수단이 필요하다. 림프관 생성 및 성장의 조절에 VEGF-C/D, VEGFR-3 pathway, podoplain이 관여된다는 사실이 밝혀져 있다. 림프관 생성 및 성장과 관련된 인자인 VEGF-C/D, podoplanin에 대한 면역조직화학염색과 반정량적 분석을 통해 미세갑상선유두상암종에서 림프절전이와의 관련성을 확인하고자 하였다. 대상 및 방법 : 2006년 9월부터 2008년 6월까지 본원에서 미세갑상선유두상암종으로 진단받고 1인 술자에 의해 갑상선 전 절제술 및 예방적 중앙 림프절절제술을 받은 104명의 환자 중 중앙 림프절 전이가 있었던 환자와 없었던 환자를 각각 25명씩 무작위로 선별하여 종양부위에 면역화학염색을 실시하여 림프관생성인자의 발현 정도를 비교하였다. 결 과 : 대상군 50예 중 VEGF-C/D는 50예(100%) 모두 발현이 되었고 podoplanin은 33예(66%)에서 발현이 되었다. 그 중 VEGF-C는 10예(20%)에서 약한 양성, 37예(74%)에서 중등도 양성, 3예(6%)에서 강한 양성소견을 보였고 VEGF-D는 9예(18%)에서 약한 양성, 37예(74%)에서 중등도 양성, 4예(8%)에서 강한 양성소견을 보였다. 중앙 림프절 전이 음성 환자 군과 양성 환자 군으로 분류하였을 때 VEGF-C/D의 발현율의 차이는 p-value가 각각 0.48, 1.00으로 통계적으로 유의한 차이를 보이지 않았다. 50예 전체를 대상으로 하여 종양의 개수, 최대크기, 검출된 전체 림프절의 수, 양성 림프절의 수, 주변조직 침범여부에 따른 VEGF-C/D의 발현도 통계적으로 유의한 차이를 보이지 않았다. Podoplanin의 경우 염색 여부에 따라 양성군과 음성군으로 나누어 분석하였을 때 종양의 개수, 최대크기, 검출된 림프절의 수, 양성 림프절의 수, 주변조직 침범여부도 통계적으로 유의한 차이를 보이지 않았다. 결 론 : VEGF-C/D는 대상군 전체(100%)에서 발현이 되었고 podoplanin은 66%에서 발현이 되었다. 림프관 생성인자로 알려진 VEGF-C/D및 podoplanin이 미세갑상선유두상암종에서 많이 발현이 되는 것으로 보아 위 인자들이 림프절 전이를 일으키는 인자 중 하나로 생각된다. 하지만 미세갑상선유두상암종에서 중앙 림프절 전이를 예측할 수 있는 인자로 부적합 한 것으로 생각되며 향후 더 많은 증례를 통해 관련성 여부에 대한 연구가 필요하고 또 다른 인자의 관련성에 대해서 연구가 필요하겠다.

• Neonatal Medicine
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• 제15권1호
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• pp.22-31
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• 2008

목 적 : 기관지폐이형성증은 미숙한 폐에서의 혈관과 폐포발달의 저해로 특징 질 수 있다. 폐 발달의 결과를 고려하면 줄기세포의 투여로 자가 회복기전을 이용한 폐 발달의 촉진의 가능성은 유망하고 이로 폐 기관지이 형성의 유병률과 합병증을 줄일 수 있다. 강화된 Green fluorescent protein (EGFP)를 표기인자로 표시한 줄기세포를 비 EGFP 마우스에 주사하여 생착 여부를 보고자 하였다. 방 법 : VEGFR2 억제제인 SU1498을 생후 3일된 마우스에 주사하여 폐포발달이 저해된 모형을 만들었다. 생후 4일에 $1{\times}10^6$ EGFP 양성 줄기세포를 복강 내로 주입하였다. 줄기세포를 투여한 폐의 형태학적인 분석과 면역염색을 시행 하였고, 주입된 줄기 세포의 생착을 확인하기 위해서 동일초점 현미경으로 분석하였다. 결 과 : SU1498을 주사한 신생마우스에서 폐포 표면적과 평균 폐포 용적이 감소되었다. 폐 발달이 억제된 마우스 모형에서 주입한 EGFP 양성 줄기세포가 발견 되었고, 내피세포와 외피세포로 분화함을 공초점 현미경으로 확인하였다. 결 론 : 주입된 EGFP 양성 줄기세포가 혈관생성억제제를 이용하여 만든 마우스의 폐 발달 저해 모형에서 생착 하여 내피세포와 외피세포로 분화함을 확인하였다.

• Reproductive and Developmental Biology
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• 제35권1호
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• pp.9-15
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• 2011

The functional cardiovascular system is comprised of distinct mesoderm-derived lineages including endothelial cells, vascular smooth muscle cells and other mesenchymal cells. Recent studies in the human embryonic stem cell differentiation model have provided evidence indicating that these cell lineages are developed from the common progenitors such as hemangioblasts and cardiovascular progenitor cells. Also, the studies have suggested that these progenitors have a common primordial progenitor, which expresses KDR (human Flk-1, also known as VEGFR2, CD309). We demonstrate here that sustained activation of BMP4 (bone morphogenetic protein 4) in hESC line, CHA15 hESC results in $KDR^+$ mesoderm specific differentiation. To determine whether the $KDR^+$ population derived from hESCs enhances potential to differentiate along multipotential mesodermal lineages than undifferentiated hESCs, we analyzed the development of the mesodermal cell types in human embryonic stem cell differentiation cultures. In embryoid body (EB) differentiation culture conditions, we identified an increased expression of $KDR^+$ population from BMP4-stimulated hESC-derived EBs. After induction with additional growth factors, the $KDR^+$ population sorted from hESCs-derived EBs displays mesenchymal, endothelial and vascular smooth muscle potential in matrix-coated monolayer culture systems. The populations plated in monolayer cultures expressed increased levels of related markers and exhibit a stable/homologous phenotype in culture terms. In conclusion, we demonstrate that the $KDR^+$ population is stably isolated from CHA15 hESC-derived EBs using BMP4 and growth factors, and sorted $KDR^+$ population can be utilized to generate multipotential mesodermal progenitors in vitro, which can be further differentiated into cardiovascular specific cells.