• 한국미생물·생명공학회지
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• 제25권2호
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• pp.137-143
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• 1997

Porcine transforming growth factor-$\beta$1 (TGF-$\beta$1) was expressed in Escherichia coli using cDNA of TGF-$\beta$1 and glutathione S-transferase (GST) fusion vector pGEX-1$\lambda$T. An ApoI-Tth111I fragment of cDNA which correspond to the amino acid residues from 123 to 390 of the precursor TGF-$\beta$1 was inserted into EcoRI-Tth111I digested pGEM#-l$\lambda$T and the recombined plasmid was named pGET-12. Gene products from the cloned regions of the recombinant plasmids pGET-12 was not detected in soluble fraction of cell free extract but detected in insoluble fraction. The solubilization of insoluble gene product was achieved by the treatment of N-laurylsarcosine. Molecular weight of partially purified proteins determined by electrophoresis was same as expected from cloned fragment. The ELISA test results of the purified proteins showed that immunologically detectable epitope was preserved in recombinant protein.

• 한국수산과학회지
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• 제49권6호
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• pp.807-814
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• 2016

Fucoidan, one of the dominant sulfated polysaccharides extracted from brown seaweed, possesses a wide range of biological activities. Transforming growth $factor-{\beta}$ ($TGF-{\beta}$) plays a pivotal role in the pathogenesis of pulmonary fibrosis, by stimulating the synthesis of profibrotic factors. In this study, we investigated the in vitro effects of fucoidan on collagen synthesis, ${\alpha}-smooth$ muscle actin (${\alpha}-SMA$) expression, and interleukin (IL)-6 production in $TGF-{\beta}$-stimulated human pulmonary fibroblasts. The expression of type I collagen and ${\alpha}-SMA$ was detected by Western blot, and the production of IL-6 by enzyme-linked immunosorbent assay. $TGF-{\beta}1$ treatment of pulmonary fibroblasts enhanced the expression of ${\alpha}-SMA$, type I collagen, and IL-6 whereas these effects were inhibited in cells pretreated with fucoidan. The activation of Smad2/3, p38 mitogen-activated protein kinases (MAPKs), and Akt was also inhibited in fucoidan-pretreated, $TGF-{\beta}1-stimulated$ human pulmonary fibroblasts. These data demonstrate the anti-fibrotic potential of fucoidan in $TGF-{\beta}-induced$ human pulmonary fibroblasts, via the inhibition of Smad2/3, p38 MAPKs, and Akt phosphorylation. Our results suggest the therapeutic potential of fucoidan in the prevention or treatment of pulmonary fibrosis.

• 대한치과교정학회지
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• 제28권2호
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• pp.311-327
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• 1998

본 연구의 목적은 시험관내에서 TGF-${\beta}$가 치주인대세포의 증식에 미치는 영향을 관찰하고, 치주 인대내에 TGF-${\beta}$ 를 투여하여 치주 인대조직에 대한 TGF-${\beta}$의 생물학적 역할을 알아보고자 시험관내에서 배양된 사람의 치주인대세포에 0.1, 1, 5, 10ng/ml농도의 TGF-${\beta}$를 투여하여 1,2,3일간 배양한후 MTT방법으로 세포의 활성을 관찰하였고, 백서의 실험적 이동시 압박측 및 견인측의 치주인대내에 TGF-${\beta}$를 투여하여 치주인대조직의 조직병리학적 관찰을 하여 다음과 같은 결과를 얻었다. 1. 0.1ng/ml 농도의 TGF-${\beta}$를 치주인대세포에 투여한 경우, 배양 1, 2, 3일째에 대조군과 유의한 차이가 없었다. 2. 1ng, 5ng/ml 농도의 TGF-${\beta}$를 치주인대세포에 투여한 경우, 배 양 1, 2일째에는 대조군과 유의한 차이가 없었으나 3일째에는 유의하게 증가되었다. 3. 10ng/m1 농도의 TGF-${\beta}$를 치주인대세포에 투여한 경우, 배양 1일째에는 대조군과 유의한 차이가 없었으나 2, 3일째에는 유의하게 증가하였다. 4. 실험적 치아이동시, TGF-${\beta}$ 투여군에서 3일째까지는 압박측에서의 초자양변성이 대조군에 비해 적었으나 7일째 이후에는 군 간의 차이가 없었고, 파골세포 출현 및 모세혈관 증식은 7일째까지 대조군보다 많이 관찰되었다. 5. 실험적 치아이동시, TGF-${\beta}$ 투여군에서 3일째부터 14일째까지 견인측의 골모세포 활성 및 신생골 형성이 증가되었던 점이 대조군과 구별되었다.

• Journal of Oral Medicine and Pain
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• 제25권1호
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• pp.53-62
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• 2000

Bone marrow culture systems are widely used to differentiate osteoclast-like cells in vitro using several osteotropic hormones. In this study, we isolated and cultured the mouse bone marrow cells with or without some osteotropic hormones such as parathyroid hormone(PTH), prostaglandin $E_2(PGE_2)$ and $l,25(OH)_2-vitamin$ $D_3$(Vit. $D_3$). We confirmed the formation of osteoclast-like cells morphologically and functionally by the expression of tartrate-resistant acid phosphatase(TRAP) and by their capability to resorb dentin slices. We also studied the effects of transforming growth $factor-{\beta}(TGF-{\beta})$ and epidermal growth factor(EGF) on the Vit. $D_3-induced$ osteoclast-like cell formation. In control, a few multinucleated cells were formed whereas PTH and $PGE_2$ increased the number of multinucleated cells. PTH, $PGE_2$ and Vit. $D_3$ induced the formation of TRAP-positive multinucleated cells. After culture of mouse bone marrow cells on the dentin slices with or without osteotropic hormones, giant cells with diverse morphology were found on the dentin slices under the scanning electronmicroscopy. After removing the attached cells, resorption pits were identified on the dentin slices, and the shape of resorption pits was variable. EGF increased the osteoclast-like cell formation induced by Vit. $D_3$, however, $TGF-{\beta}$ showed biphasic effect, which at low concentration, increased and at high concentration, decreased the osteoclast-like cell formation induced by Vit. $D_3$.

• Journal of Ginseng Research
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• 제43권1호
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• pp.68-76
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• 2019

Background: In colorectal cancer (CRC), 40-60% of patients develop metastasis. The epithelial-mesenchymal transition (EMT) is a pivotal and intricate process that increases the metastatic potential of CRC. The aim of this study was to investigate the effect of Korean Red Ginseng extract (RGE) on colorectal metastasis through inhibition of EMT and the metastatic abilities of CRC cells. Methods: To investigate the effect of RGE on the metastatic phenotypes of CRC cells, CT26 and HT29 cells were evaluated by using an adhesion assay, a wound-healing assay, an invasion assay, zymography, and real-time reverse transcription-polymerase chain reaction. Western-blot analysis was conducted to elucidate the molecular mechanisms of RGE, which showed an inhibitory effect on the transforming growth factor-${\beta}1$ ($TGF-{\beta}1$)-induced EMT in HT29 cells. Additionally, the antimetastatic effect of RGE was evaluated in a mouse model of lung metastasis injected with CT26 cells. Results: RGE decreased the adhesion and migration ability of the CT26 cells and TGF-${\beta}1$-treated HT29 cells. The invasion ability was also reduced by RGE treatment through the inhibition of matrix metalloproteinase-9 expression and activity. Moreover, RGE suppressed the TGF-${\beta}1$-induced EMT via TGF-${\beta}1$/Smad-signaling-mediated Snail/E-cadherin expression in HT29 cells and lung tissue in CT26 tumor-bearing mice. Conclusion: Our results demonstrated that RGE inhibited colorectal lung metastasis through a reduction in metastatic phenotypes, such as migration, invasion, and the EMT of CRC cells.

• International Journal of Molecular Medicine
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• 제44권2호
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• pp.491-502
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• 2019

Although the migration of hepatic stellate cells (HSCs) is important for hepatic fibrosis, the regulation of this migration is poorly understood. Notably, transforming growth factor (TGF)-β1 induces monocyte migration to sites of injury or inflammation during the early phase, but inhibits cell migration during the late phase. In the present study, the role of transforming protein RhoA signaling in TGF-β1-induced HSC migration was investigated. TGF-β1 was found to increase the protein and mRNA levels of smooth muscle actin and collagen type I in HSC-T6 cells. The level of RhoA-GTP in TGF-β1-stimulated cells was significantly higher than that in control cells. Furthermore, the phosphorylation of cofilin and formation of filamentous actin (F-actin) were more marked in TGF-β1-stimulated cells than in control cells. Additionally, TGF-β1 induced the activation of nuclear factor-κB, and the expression of extracellular matrix proteins and several cytokines in HSC-T6 cells. The active form of Rap1 (Rap1 V12) suppressed RhoA-GTP levels, whereas the dominant-negative form of Rap1 (Rap1 N17) augmented RhoA-GTP levels. Therefore, the data confirmed that Rap1 regulated the activation of RhoA in TGF-β1-stimulated HSC-T6 cells. These findings suggest that TGF-β1 regulates Rap1, resulting in the suppression of RhoA, activation of and formation of F-actin during the migration of HSCs.

• Asian-Australasian Journal of Animal Sciences
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• 제16권3호
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• pp.335-341
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• 2003

Carcass weight and backfat thickness are two of important elements in determining the carcass trait in pigs and are studied on animal genetics, nutrition, and endocrinology. Growth factors stimulate or inhibit the proliferation and differentiation of various cells. In particular, insulin-like growth factors (IGFs), transforming growth factor (TGF)-$\beta$, and epidermal growth factor (EGF) are involved in the growth and maintenance of muscle. Also, dehydroepiandrosterone-sulfate (DHEA-S) and cortisol are known to be related to the obesity and subcutaneous fat depth in pigs. Therefore, this study was performed to relate growth factors (IGFs, TGF-${\beta}1$, and EGF) and hormones (cortisol and DHEA-S) concentrations at antemortem and postmortem periods to carcass traits including carcass weight and backfat thickness. Blood and m. Longissimus were collected in pigs at antemortem (30 days before slaughter) and postmortem periods. After slaughtered, carcass weight and backfat thickness were measured. Growth factors and hormones in serum and m. Longissimus were measured by radioimmunoassay or enzyme-linked imuunosorbent assay. Before antemortem period, serum IGF-I and -II concentrations were positively correlated with the carcass weight and backfat thickness in gilts, and the concentrations of TGF- ${\beta}1$ and cortisol in barrows show the correlation with only carcass weight. Also, the positive correlations of muscular IGFs and TGF-${\beta}1$ at postmortem 45 min with the carcass weight and backfat thickness were detected. Consequently, these results suggest that the serum and muscular endocrine factors are involved in the carcass weight and backfat thickness in pigs.

• Toxicological Research
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• 제27권4호
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• pp.253-259
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• 2011

Transforming growth factor ${\beta}$ (TGF-${\beta}$) is involved in cellular processes including growth, differentiation, apoptosis, migration, and homeostasis. Generally, TGF-${\beta}$ is the inhibitor of cell cycle progression and plays a role in enhancing the antagonistic effects of many growth factors. Unlike the antiproliferative effect of TGF-${\beta}$, E2, an endogeneous estrogen, is stimulating cell proliferation in the estrogen-dependent organs, which are mediated via the estrogen receptors, $ER{\alpha}$ and $ER{\beta}$, and may be considered as a critical risk factor in tumorigenesis of hormone-responsive cancers. Previous researches reported the cross-talk between estrogen/$ER{\alpha}$ and TGF-${\beta}$ pathway. Especially, based on the E2-mediated inhibition of TGF-${\beta}$ signaling, we examined the inhibition effect of 4-tert-octylphenol (OP) and 4-nonylphenol (NP), which are well known xenoestrogens in endocrine disrupting chemicals (EDCs), on TGF-${\beta}$ signaling via semi-quantitative reverse-transcription PCR. The treatment of E2, OP, or NP resulted in the downregulation of TGF-${\beta}$ receptor2 (TGF-${\beta}$ R2) in TGF-${\beta}$ signaling pathway. However, the expression level of TGF-${\beta}1$ and TGF-${\beta}$ receptor1 (TGF-${\beta}$ R1) genes was not altered. On the other hand, E2, OP, or NP upregulated the expression of a cell-cycle regulating gene, c-myc, which is a oncogene and a downstream target gene of TGF-${\beta}$ signaling pathway. As a result of downregulation of TGF-${\beta}$ R2 and the upregulation of c-myc, E2, OP, or NP increased cell proliferation of BG-1 ovarian cancer cells. Taken together, these results suggest that E2 and these two EDCs may mediate cancer cell proliferation by inhibiting TGF-${\beta}$ signaling via the downregulation of TGF-${\beta}$ R2 and the upregulation of c-myc oncogene. In addition, it can be inferred that these EDCs have the possibility of tumorigenesis in estrogen-responsive organs by certainly representing estrogenic effect in inhibiting TGF-${\beta}$ signaling.

• Molecules and Cells
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• 제24권3호
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• pp.431-436
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• 2007

Stem cell-based therapy is being considered as an alternative treatment for cardiomyopathy. Hence understanding the basic molecular mechanisms of cardiomyocyte differentiation is important. Besides BMP or Wnt family proteins, $TGF-{\beta}$ family members are thought to play a role in cardiac development and differentiation. Although $TGF-{\beta}$ has been reported to induce cardiac differentiation in embryonic stem cells, the differential role of $TGF-{\beta}$ isoforms has not been elucidated. In this study, employing the DMSO-induced cardiomyocyte differentiation system using P19CL6 mouse embryonic teratocarcinoma stem cells, we investigated the $TGF-{\beta}$-induced signaling pathway in cardiomyocyte differentiation. $TGF-{\beta}1$, but not the other two isoforms of $TGF-{\beta}$, was induced at the mRNA and protein level at an early stage of differentiation, and Smad2 phosphorylation increased in parallel with $TGF-{\beta}1$ induction. Inhibition of $TGF-{\beta}1$ activity with $TGF-{\beta}1$-specific neutralizing antibody reduced cell cycle arrest as well as expression of the CDK inhibitor $p21^{WAF1}$. The antibody also inhibited induction of the cardiac transcription factor Nkx2.5. Taken together, these results suggest that $TGF-{\beta}1$ is involved in cardiomyocyte differentiation by regulating cell cycle progression and cardiac gene expression in an autocrine or paracrine manner.

• Journal of Animal Science and Technology
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• 제55권2호
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• pp.95-101
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• 2013

The present study was conducted to test a hypothesis that pork quality traits would be influenced by the systemic and/or local bioavailability of insulin-like growth factor-I (IGF-I), transforming growth factor-${\beta}1$ (TGF-${\beta}1$), or epidermal growth factor (EGF) before or at slaughter. To this end, 60 cross-bred female market pigs weighing approximately 110 kg were slaughtered, after which Longissimus dorsi muscle (LM) samples taken at slaughter (D 0) and blood samples taken at D -7 and D 0 were analyzed. The 60 carcasses rendered 36 RFN (reddish-pink, firm, and non-exudative), 16 RSE (reddish-pink, soft, and exudative), and 6 PSE (pale, soft, and exudative); 2 DFD (dark, firm, and dry) also were found but were excluded in subsequent experiments. The $L^*$ and drip loss were greater in PSE vs. RFN and RSE and in PSE and RSE vs. RFN, respectively, as they should (P<0.05). The $pH_{45min}$ was less in PSE vs. RFN (P<0.05); $pH_{24h}$ tended to be less in the former (P=0.09). The LM IGF-I and TGF-${\beta}1$ as well as serum EGF concentrations were less in PSE than in RFN. None of the other LM and serum concentrations of the three growth factors differed across the three pork quality categories. The LM IGF-I and TGF-${\beta}1$ concentrations and serum EGF concentration at D 0 were negatively correlated with drip loss [r = -0.36(P<0.01), -0.44 (P<0.01), and -0.32 (P<0.05), respectively]. However, none of the serum and LM growth factor variables was correlated with $L^*$ or $a^*$ (redness) of LM. Taken together, results suggest that locally expressed IGF-I and TGF-${\beta}1$ and blood-borne EGF may have a beneficial effect on postmortem water holding capacity of the muscle and that pork quality traits could be predicted to some extent from concentrations of IGF-I and TGF-${\beta}1$ in muscle and EGF in serum at slaughter.