• Asian Pacific Journal of Cancer Prevention
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• 제15권5호
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• pp.2291-2295
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• 2014

Although nucleolar protein nucleostemin (NS) is essential for cell proliferation and early embryogenesis and expression has been observed in some types of human cancer and stem cells, the molecular mechanisms involved in mediation of cell proliferation and cell cycling remains largely elusive. The aim of the present study was to evaluate NS as a potential target for gene therapy of human breast carcinoma by investigating NS gene expression and its effects on SKBR-3 cell proliferation and apoptosis. NS mRNA and protein were both found to be highly expressed in all detected cancer cell lines. The apoptotic rate of the pcDNA3.1-NS-Silencer group ($12.1-15.4{\pm}3.8%$) was significantly higher than those of pcDNA3.1-NS ($7.2-12.0{\pm}1.7%$) and non-transfection groups ($4.1-6.5{\pm}1.8%$, P<0.01). MTT assays showed the knockdown of NS expression reduced the proliferation rate of SKBR-3 cells significantly. Matrigel invasion and wound healing assays indicated that the number of invading cells was significantly decreased in the pcDNA3.1-NS-siRNA group (P<0.01), but there were no significant difference between non-transfected and over-expression groups (P>0.05). Moreover, RNAi-mediated NS down-regulation induced SKBR-3 cell G1 phase arrest, inhibited cell proliferation, and promoted p53 pathway-mediated cell apoptosis in SKBR-3 cells. NS might thus be an important regulator in the G2/M check point of cell cycle, blocking SKBR-3 cell progression through the G1/S phase. On the whole, these results suggest NS might be a tumor suppressor and important therapeutic target in human cancers.

• Journal of Nutrition and Health
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• 제41권1호
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• pp.13-21
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• 2008

본 실험은 인돌의 인체 대장암세포의 경과 및 전이억제와 TJ 활성 조절에 미치는 영향을 알아보기 위해 실험하였다. 인돌은 십자화 야채류인 양배추, 컬리플라워, 브로클리 등에 존재하는 glucosinolate, glucobrassicin의 대사산물 이다. 본 연구의 결과는 인돌이 대장암 세포 HT-29에서 농도 의존적으로 세포증식 저해를 나타내었다. 상처회복 실험을 통한 세포이동성과 세포 침윤성 실험결과 인돌이 암세포의 이동과 침윤성을 억제하였고 투과성상피세포의 전기적 저항성 측정치는 인돌 처리 세포에서 증가하였다. HT-29 세포에서 과발현을 나타내는 밀착결합 단백질인 claudin-1, claudin-5 발현은 인돌 처리로 유전자의 전사수준과 단백질 수준에서 유의적인 감소를 나타내었다. 이상의 결과에서 인돌이 HT-29 세포의 밀착결합과 그 구성 단백질 중 claudin 발현 현상을 회복시키므로 암 경과와 전이 억제를 나타내었다. 결론적으로, 천연 항암화합물인 인돌은 대장암 세포 HT-29에서 밀착결합 단백질인 claudin-1, -5을 억제하여 밀착결합을 활성화하므로 암 진행과 전이를 억제할 수 있는 인돌에 의한 새로운 기전을 보고합니다.

• 동의생리병리학회지
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• 제16권3호
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• pp.452-457
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• 2002

Galla Rhois is a gallnut of Rhus javanica Linne used for treatment of diarrhea, hemorrhage, cough, leukorrhea and toxic tumor etc in oriental medicine. For the evaluation of antitumor effect of Galla Rhois, activity based fractionation was done. We isolated an effective compound and identified 1,2,3,4,6-penta-O-galloyl-β-D-glucose(PGG) by photometric analysis such as NMR and MASS. Then, we studied the angiogenic activity of PGG. It showed a cytotoxicity against SK-OV-3, SK-OV-3, HT1080 with IC/sub 50/ of 50 ug/ml approximately. It also effectively inhibited proliferation of HUVEC cells treated by bFGF to 30% of control at 20 ug/ml and cell migration to 80% at 10 ug in a dose dependent fashion. Tube formation of HUVEC cells on matrigel was effectively suppressed from 2.5 ug/ml of concentration by PGG. Moreover, it effectively recovered the dysfunction of gap junctional intercellular communication in WB-F344 rat liver epithelial cells caused by hydrogen peroxide at 4 ug/ml suggesting it potently can inhibit tumor promotion. Taken together, it indicates 1,2,3,4,6-penta-O-galloyl- β -D-glucose has antiangiogenic activity.

• 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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• 제23권1호
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• pp.131-136
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• 2013

Sirtuin family 단백질은 암, 당뇨, 심혈관 질환, 뇌신경계 질환과 같은 노화와 연관된 질병들의 발병에 중요한 역할을 하는 것으로 알려져 있다. 우선적으로 혈관내피세포를 이용하여 Hypoxia에서의 sirtuin family의 발현을 확인한 결과, SIRT2 mRNA가 가장 강하게 발현되는 결과를 얻었다. 혈관신생과정에서의 SIRT2 단백질의 생리학적 역할을 규명하고자, 본 실험실에서는 저산소상태에서의 SIRT2의 생리학적인 의미에 주안점을 두었다. Normoxia에서 SIRT2는 세포질에 존재하고 있으나, hypoxia에 의해서 SIRT2 단백질이 핵 내로 이동이 일어남을 확인하였다. 또한 hypoxia에 의해서 SIRT2와 혈관신생인자인 VEGF의 발현이 증가하며, Normoxia와 hypoxia 두 환경에서 혈관내피세포의 혈관형성능력이 SIRT2 inhibitor인 AK-1에 의해서 억제된 것을 확인하였다. 본 연구결과를 통해서 SIRT2가 혈관신생과정을 조절하는 중요한 역할을 함을 시사하였다.

• Clinical and Experimental Reproductive Medicine
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• 제47권2호
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• pp.114-121
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• 2020

Objective: Despite extensive research on implantation failure, little is known about the molecular mechanisms underlying the crosstalk between the embryo and the maternal endometrium, which is critical for successful pregnancy. Profilin 1 (PFN1), which is expressed both in the embryo and in the endometrial epithelium, acts as a potent regulator of actin polymerization and the cytoskeletal network. In this study, we identified the specific role of endometrial PFN1 during embryo implantation. Methods: Morphological alterations depending on the status of PFN1 expression were assessed in PFN1-depleted or control cells grown on Matrigel-coated cover glass. Day-5 mouse embryos were cocultured with Ishikawa cells. Comparisons of the rates of F-actin formation and embryo attachment were performed by measuring the stability of the attached embryo onto PFN1-depleted or control cells. Results: Depletion of PFN1 in endometrial epithelial cells induced a significant reduction in cell-cell adhesion displaying less formation of colonies and a more circular cell shape. Mouse embryos co-cultured with PFN1-depleted cells failed to form actin cytoskeletal networks, whereas more F-actin formation in the direction of surrounding PFN1-intact endometrial epithelial cells was detected. Furthermore, significantly lower embryo attachment stability was observed in PFN1-depleted cells than in control cells. This may have been due to reduced endometrial receptivity caused by impaired actin cytoskeletal networks associated with PFN1 deficiency. Conclusion: These observations definitively demonstrate an important role of PFN1 in mediating cell-cell adhesion during the initial stage of embryo implantation and suggest a potential therapeutic target or novel biomarker for patients suffering from implantation failure.

• Molecules and Cells
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• 제41권6호
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• pp.515-522
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• 2018

Patients with head and neck cancer are treated with therapeutic irradiation, which can result in irreversible salivary gland dysfunction. Because there is no complete cure for such patients, stem cell therapy is an emerging alternative for functional restoration of salivary glands. In this study, we investigated in vitro characteristics of primarily isolated epithelial cells from human salivary gland (Epi-SGs) and in vivo formation of acini-like structures by Epi-SGs. Primarily isolated Epi-SGs showed typical epithelial cell-like morphology and expressed E-cadherin but not N-cadherin. Epi-SGs expressed epithelial stem cell (EpiSC) and embryonic stem cell (ESC) markers. During long-term culture, the expression of EpiSC and ESC markers was highly detected and maintained within the core population with small size and low cytoplasmic complexity. The core population expressed cytokeratin 7 and cytokeratin 14, known as duct markers indicating that Epi-SGs might be originated from the duct. When Epi-SGs were transplanted in vivo with Matrigel, acini-like structures were readily formed at 4 days after transplantation and they were maintained at 7 days after transplantation. Taken together, our data suggested that Epi-SGs might contain stem cells which were positive for EpiSC and ESC markers, and Epi-SGs might contribute to the regeneration of acini-like structures in vivo. We expect that Epi-SGs will be useful source for the functional restoration of damaged salivary gland.

• 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).

• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권4호
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• pp.299-305
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• 2010

Purpose: Adipose tissue is located beneath the skin, around internal organs, and in the bone marrow in humans. Its main role is to store energy in the form of fat, although it also cushions and insulates the body. Adipose tissue also has the ability to dynamically expand and shrink throughout the life of an adult. Recently, it has been shown that adipose tissue contains a population of adult multipotent mesenchymal stem cells and endothelial progenitor cells that, in cell culture conditions, have extensive proliferative capacity and are able to differentiate into several lineages, including, osteogenic, chondrogenic, endothelial cells, and myogenic lineages. Materials and Methods: This study focused on endothelial cell culture from the adipose tissue. Adipose tissues were harvested from buccal fat pad during bilateral sagittal split ramus osteotomy for surgical correction of mandibular prognathism. The tissues were treated with 0.075% type I collagenase. The samples were neutralized with DMEM/and centrifuged for 10 min at 2,400 rpm. The pellet was treated with 3 volume of RBC lysis buffer and filtered through a 100 ${\mu}m$ nylon cell strainer. The filtered cells were centrifuged for 10 min at 2,400 rpm. The cells were further cultured in the endothelial cell culture medium (EGM-2, Cambrex, Walkersville, Md., USA) supplemented with 10% fetal bovine serum, human EGF, human VEGF, human insulin-like growth factor-1, human FGF-$\beta$, heparin, ascorbic acid and hydrocortisone at a density of $1{\times}10^5$ cells/well in a 24-well plate. Low positivity of endothelial cell markers, such as CD31 and CD146, was observed during early passage of cells. Results: Increase of CD146 positivity was observed in passage 5 to 7 adipose tissue-derived cells. However, CD44, representative mesenchymal stem cell marker, was also strongly expressed. CD146 sorted adipose tissue-derived cells was cultured using immuno-magnetic beads. Magnetic labeling with 100 ${\mu}l$ microbeads per 108 cells was performed for 30 minutes at $4^{\circ}C$ a using CD146 direct cell isolation kit. Magnetic separation was carried out and a separator under a biological hood. Aliquous of CD146+ sorted cells were evaluated for purity by flow cytometry. Sorted cells were 96.04% positivity for CD146. And then tube formation was examined. These CD146 sorted adipose tissue-derived cells formed tube-like structures on Matrigel. Conclusion: These results suggest that adipose tissue-derived cells are endothelial cells. With the fabrication of the vascularized scaffold construct, novel approaches could be developed to enhance the engineered scaffold by the addition of adipose tissue-derived endothelial cells and periosteal-derived osteoblastic cells to promote bone growth.