• Title/Summary/Keyword: Epithelial growth

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Regulation of the plasminogen activator activity and inflammatory environment via transforming growth factor-beta regulation of sperm in porcine uterine epithelial cells

  • Kim, Su-jin;Cheong, Hee-Tae;Park, Choon-keun
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.4
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    • pp.297-306
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    • 2020
  • The aims of the present study were to confirm that regulation of the PA and environment via TGF-β regulation of sperm by Percoll-separated in porcine uterine epithelial cells. And, it was performed to identify the cytokines (TGF-β1, 2 and 3, TGF-β receptor1 and 2; interleukin, IL-6, IL-8) and PA-related genes (urokinase-PA, uPA; tissue-PA, tPA; PA inhibitor, PAI; uPA-receptor, uPAR) by spermatozoa. The experiment used porcine uterus epithelial cells (pUECs) and uterine tissue epithelial cells, Boar sperm were separated by discontinuous Percoll density gradient (45/90%), and tissues were co-incubated with spermatozoa, followed by real-time PCR. PA activity was measured of sperm by discontinuous Percoll density gradient (45/90%) for 24 hours. To measure viability and acrosome damage of sperm double stained propidium iodide (PI) and SYBR-14 or FITC-PNA were used. In results, binding ratio of Percoll-separated sperm was found no differences, but sperms isolated from 90% Percoll layer reduced PA activity (p < 0.05). when co-cultured sperm selected Percoll in porcine uterus tissues epithelial cells, 90% layer sperm increased TGF-β R1, contrastively tPA and PAI-1 in comparison with control (p < 0.05). 45% sperm was decreased the expression of uPA (p < 0.05). TGF-β decreased PA activity in the supernatant collected from pUECs (p < 0.05). Especially, The group including uPA, PAI-1 were induce sperm intact, while it was reduced in sperm damage when compared to control (p < 0.05). Also, there was no significant difference group of tPA and tPA+I in the dead sperm and acrosome damage compared to control. The expression of tPA and PAI showed a common response. Percoll-separated spermatozoa in 90% layer reduced tPA and IL-related gene mRNA expression. Thus, Percoll-sparated sperm in 90% layer show that it can suppress inflammation through increased expression of TGF-β and downregulation of PA and IL in epithelial cells compared to 45% layer Percoll.

Epithelial to Mesenchymal Transition in CAPD Patients (복막의 섬유화 기전으로서의 상피중간엽전이)

  • Do, Jun-Young
    • Journal of Yeungnam Medical Science
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    • v.23 no.1
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    • pp.10-18
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    • 2006
  • Epithelial to mesenchymal transition (EMT) is an important etiologic factor for the development of peritoneal fibrosis in CAPD patients. Mesothelial cells are main source of trans-differentiated fibroblasts under stress from the bioincompatible peritoneal dialysate. In our study there was no difference in dialysate TGF-${\beta}$ and VEGF between the low and high GDP groups during an initial 12 months. However, after adjusting with D-CA125, the low GDPs group showed a significantly lower D-TGF-${\beta}$/D-CA125 and D-VEGF/CA125 during the initial 12 months. Among the adjusted peritoneal growth factors for CA125, VEGF/CA125 and TGF-b/CA125 were factors significantly associated with greater EMT in this study. Adjustment of the peritoneal growth factor for effluent CA125 (surrogate for mass of HPMCs) revealed significant association with EMT suggesting that the fibroblastoid transition from HPMCs could be affected by the amount of intraperitoneal growth factors (TGF-b, VEGF) per unit mass of HPMCs. There was significant improvement in both cell score and D-CA125 at the sixth and 12th months after switching from a high GDPs solution to a low GDPs solution. Use of icodextrin solution in patients who had average peritoneal transport showed not only better systemic effects such as decreased glucose absorption via dialysate but also preservation of the peritoneum, including less EMT and high mesothelial bulk mass. In conclusion, Therapy with low GDP solution including icodextrin may positively impact preservation of the peritoneal membrane integrity and prevention of peritoneal fibrosis with time on PD.

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ATP and Purinergic Receptor Agonists Stimulate the Mitogen-Activated Protein Kinase Pathway and DNA Synthesis in Mouse Mammary Epithelial Cells

  • Yuh In-Sub
    • Reproductive and Developmental Biology
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    • v.28 no.4
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    • pp.211-219
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    • 2004
  • The effects of adenosine 5'-triphosphate (ATP) and ATP analogs, P/sub 2y/ purinoceptor agonists, on growth of normal mouse mammary epithelial cells (NMuMG) were examined. Cells were plated onto 24 well plates in DMEM supplemented with 10 % fetal calf serum. After serum starvation for 24 hours, ATP, P/sub 2y/ purinoceptor agonists (AdoPP[NH]P, ATP-α-S, ATP-γ-S, β, γ-me-ATP and 2me-S-ATP), P/sub 2u/ purinoceptor agonist (UTP) and P/sub 2y/ purinoceptor antagonists (Reactive Blue 2, more selective to P/sub 2y/ receptor than PPADS; PPADS) were added. DNA synthesis was estimated as incorporation of 3H-thymidine into DNA (1 hour pulse with 1 μ Ci/ml, 18~19 hours after treatment). ATP, Adopp[NH]P, ATP-α-S or ATP-γ-S, significantly increased DNA synthesis at 1, 10 and 100 μM concentrations with dose-dependency (P<0.05), and the maximum responses of ATP and ATP analogs were shown at 100 μM concentration (P<0.05). The potency order of DNA synthesis was ATP≥ATP- γ -S>Adopp [NH]P>ATP-α-S. β, γ -me-ATP, 2me-S-ATP and UTP did not increase DNA synthesis. In autoradiographic analysis of percentage of S-phase cells, similar results were observed to those of DNA synthesis. Addition of 1, 10 or 100 μM Reactive Blue 2 or PPADS significantly decreased ATP (100 μM)-induced DNA synthesis, however, PPADS was less effective than Reactive Blue 2. In Elvax 40P implant experiment, ATP directly stimulated mammary endbud growth in situ suggesting the physiological regulator of ATP in mammary growth. ATP 100 μM rapidly increased MAPK activity, reaching a maximum at 5 min and then gradually decreasing to the base level in 30 min. ATP analogs, Adopp[NH]P and ATP-γ-S also increased MAPK activity, however, β, γ-me-ATP and 2me-S-ATP did not. The inhibitor of the upstream MAPK kinase (MEK), PD 98059 (25 μM), effectively reduced ATP (100 μM) or EGF(10 ng/ml, as positive control)-induced MAPK activity and DNA synthesis (P<0.05). These results indicate that ATP-induced DNA synthesis was prevented from the direct inhibition of MAPK kinase pathway. Overall results support the hypothesis that the stimulatory effects of normal mouse mammary epithelial growth by addition of ATP or ATP analogs are mediated through mammary tissue specific P/sub 2y/ purinoceptor subtype, and MAPK activation is necessary for the ATP-induced cell growth.

Characterization and Immortalization of Normal Human Endometrial Cells

  • Hong, In-Sun;Kim, Seok-Hyun;Kung, Mi-Kyung;Jun, Jin-Hyun;Lee, Yong-Soon;Kang, Kyung-Sun
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2003.10b
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    • pp.122-122
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    • 2003
  • Endometrial tissue is an interesting model for intrinsic and extrinsic factors, ie hormones and growth factors, involved in its normal pathologic development and its cyclic growth. The endometrial cells were isolated from endometrial tissue of the proliferative phase obtained by hysterectomy and separated stromal and epithelial cells.(omitted)

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Effects of Kimchi Extract on the Development of Multicellular Structures from Rat Mammary Organoids Cultured in Matrigel

  • Kim, Nam-Deuk;Hur, Young-Mi;Rhee, Sook-Hee;Park, Kun-Young
    • Preventive Nutrition and Food Science
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    • v.1 no.2
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    • pp.168-173
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    • 1996
  • The effect of methanol soluble fraction(MSF) of kimchi on the proliferating and differentiating activity of normal rat mammary epithelial cells or organoids in culture were studied. Reconstituted basement membrane, Matrigel, supported the growth and development several different multicellular structures from mammary organoids. The five type colonies of multicellular structures, stellate, ductal, webbed, squamous, and lobulo-ductal colonies, were observed in Matrigel culture. In methanol extract groups, webbed colonies were more and squamous colonies were less than control group. and the lobulo-ductal colonies which is known that it formed in well differentiated mammary epoithelial cells were developed more in MSF treated group than control group. These results showed that methanol extract of kimchi affected on the proliferation and differentiation of normal rat mammary epithelial cells cultured in serum free medium condition.

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Generation and analysis of whole-genome sequencing data in human mammary epithelial cells

  • Jong-Lyul Park;Jae-Yoon Kim;Seon-Young Kim;Yong Sun Lee
    • Genomics & Informatics
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    • v.21 no.1
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    • pp.11.1-11.5
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    • 2023
  • Breast cancer is the most common cancer worldwide, and advanced breast cancer with metastases is incurable mainly with currently available therapies. Therefore, it is essential to understand molecular characteristics during the progression of breast carcinogenesis. Here, we report a dataset of whole genomes from the human mammary epithelial cell system derived from a reduction mammoplasty specimen. This system comprises pre-stasis 184D cells, considered normal, and seven cell lines along cancer progression series that are immortalized or additionally acquired anchorage-independent growth. Our analysis of the whole-genome sequencing (WGS) data indicates that those seven cancer progression series cells have somatic mutations whose number ranges from 8,393 to 39,564 (with an average of 30,591) compared to 184D cells. These WGS data and our mutation analysis will provide helpful information to identify driver mutations and elucidate molecular mechanisms for breast carcinogenesis.

Inhibitory Effect of Radish on Gastric Cell Toxicity and Interleukin-8 Production Induced by Helicobacter pylori (Helicobacter pylori에 의한 위세포독성 및 interleukin-8 생성에 미치는 무의 억제효과)

  • Shon Yun Hee;Suh Jeong Ill;Park In Kyung;Hwang Cher Won;Kim Cheorl Ho;Nam Kyung Soo
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.595-599
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    • 2005
  • The efforts of Korean and Japanese radishes on the viability and interleukin (JL)-8 production by Helicobacter pylori were investigated in human gastric epithelial cell. Cell viability was significantly decreased when they were incubated with H. pylori toxin (p <0.05, p<0.01 and p<0.005). Co-incubation with Korean or Japanese radish increased H. pylori toxin-inhibited cell growth in a concentration-dependent manner. The production of IL-8 was greatly increased in H. pylori-infected gastric epithelial cell in concentration- and time-dependent manners. The increased production of IL-8 was significantly inhibited by Korean (p<0.05 and p<0.01) or Japanese (p<0.05) radishes $(5\~10mg/ml)$. These results indicate that Korean and Japanese radishes have protective effects on H. pylori-inhibited cell growth and H. pylori-induced gastric mucosal cell inflammation by suppressing the production of inflammatory cytokine (IL-8) from gastric epithelial cell.

Study of Rat Mammary Epithelial Stem Cells In Vivo and In Vitro (생체 및 시험관에서 유선 상피 모세포의 분리와 동정)

  • Nam Deuk Kim;Kee-Joo Paik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.3
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    • pp.470-486
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    • 1995
  • Mammary epithelial cells contain a subpopulation of cells with a large proliferativ potential which are responsible for the maintenance of glandular cellularity and are the progenitor cells of mammary cancer. These clonogens give rise to multicellular clonal alveolar or ductal units(AU or DU) on transplantation and hormonal stimulation. To isolate putative mammary clonogens, enzymatically monodispersed rat mammary epithelial cells from organoid cultures and from intact glands are sorted by flow cytometry according to their affinity for FITC labeled peanut lectin(PNA) and PE labeled anti-Thy-1.1 antibody(Thy-1.1) into four subpopulations : cells negative to both PNA and Thy-1.1(B-), PNA+cells, Thy-1.1+cells, and cells positive to both reagents(B+). The in vivo transplantation assays indicate that the clonogenic fractions of PNA+cells from out-growths of organoids in primary cultures for three days in complete hormone medium(CHM) are significantly higher than those of cells from other subpopulations derived from cultrues or from intact glands. Extracellular matrix(ECM) is a complex of several proteins that regulated cell function ; its role in cell growth and differentiation and tissue-specific gene expression. It can act as a positive as well as a negative regulator of cellular differentiation depending on the cell type and the genes studied. Regulation by ECM is closely interrelated with the action of other regulators of cellular function, such as growth factors and hormones. Matrigel supports the growth and development of several different multicellular colonies from mammary organoids and from monodispersed epithelial cells in culture. Several types of colonies are observed including stellate colonies, duct-like structures, two- and three-dimensional web structures, squamous organoids, and lobulo-duct colonies. Organoids have the greatest proliferative potential and formation of multi-cellular structures. Phase contrast micrographs demonstrate extensive intracellular lipid accumulation within the web structures and some of duct-like colonies. At the immunocytochemical and electron micrograph level, casein proteins are predominantly localized near the apical surface of the cells or in the lumen of duct-like or lobulo-duct colonies. Squamous colonies are comprised of several layers of squamous epithelium surrounding keratin pearls as is typical fo squamous metaplasia(SM). All-trans retinoic acid(RA) inhibits the growth of SM. The frequency of lobulo-ductal colony formation increased with the augmentation of RA concentration in these culture conditions. The current study models could provide powerful tools not only for understanding cell growth and differentiation of epithelial cells, but also for the isolation and characterization of mammary clonogenic stem cells.

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Effects of the b-FGF to Early Revascularization and Epithelial Regeneration in the Rabbit's Tracheal Autograft (염기 섬유아세포 성장인자가 토끼기관의 자가이식편의 초기 혈관재형성 및 상피세포 재생에 미치는 영향)

  • 성숙환;원태희
    • Journal of Chest Surgery
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    • v.30 no.6
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    • pp.559-565
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    • 1997
  • Donor airway ischemia is a significant problem after tracheal replacement with homograft or lung transplantation, Several factors such as omentopexy, heparin, PGl2 and fibroblast growth factor, have been shown to induce angiogenesis in vitro and in vivo. This study was designed to investigate whether omentopexy and basic flbroblast growth factor can enhance rabbit tracheal revascularization and epithelial regeneration, Three different experiments were performed with New Zealand white rabbit. In group I(n= 15 control group), only coNical tracheal autotransplantation was done. In group II(n= 15), cervical tracheal autotransplantation with omentopexy was done through subcutaneous route. In group III(n= 15), cervical tracheal autotransplantation was done and lug basic flbroblast growth factor was applied. After 3, 7 and 14 days, the animals were sacrificed. The extent of revascularization was investigated by means of uptake of the human serum albumin labelled with 99m technetium, and epithelial regeneration were assessed by means of light microscope. In the group investigated at day 3, there was statistically significant high tracheal revascularization in group III(p<0.05), but no difference at 7 and 14 days. And epithelial regenerations at day 3 were better in group III(p<0.05), and at day 7 in group II and III. But there was no difference at day 14. We concluded that b-FGF can enhance the revascularization and epithelial regeneration of the tracheal autograft especially in early phase.

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Wnt5a attenuates the pathogenic effects of the Wnt/β-catenin pathway in human retinal pigment epithelial cells via down-regulating β-catenin and Snail

  • Kim, Joo-Hyun;Park, Seoyoung;Chung, Hyewon;Oh, Sangtaek
    • BMB Reports
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    • v.48 no.9
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    • pp.525-530
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    • 2015
  • Activation of the Wnt/β-catenin pathway plays a pathogenic role in age-related macular degeneration (AMD) and is thus a potential target for the development of therapeutics for this disease. Here, we demonstrated that Wnt5a antagonized β-catenin response transcription (CRT) induced with Wnt3a by promoting β-catenin phosphorylation at Ser33/Ser37/Thr41 and its subsequent degradation in human retinal pigment epithelial (RPE) cells. Wnt5a decreased the levels of vascular endothelial growth factor (VEGF), tumor necrosis factor-α(TNF-α), and nuclear factor-κB (NF-κB), which was up-regulated by Wnt3a. Furthermore, Wnt5a increased E-cadherin expression and decreased cell migration by down-regulating Snail expression, thereby abrogating the Wnt3a-induced epithelial-mesenchymal transition (EMT) in human RPE cells. Our findings suggest that Wnt5a suppresses the pathogenic effects of canonical Wnt signaling in human RPE cells by promoting β-catenin phosphorylation and degradation. Therefore, Wnt5a has significant therapeutic potential for the treatment of AMD. [BMB Reports 2015; 48(9): 525-530]