• Experimental and Molecular Medicine
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• v.50 no.12
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• pp.4.1-4.19
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• 2018

Transforming growth factor $(TGF)-{\beta}$ signaling is increasingly recognized as a key driver in cancer. In progressive cancer tissues, $TGF-{\beta}$ promotes tumor formation, and its increased expression often correlates with cancer malignancy. In this study, we utilized adenoviruses expressing short hairpin RNAs against $TGF-{\beta}1$ and $TGF-{\beta}2$ to investigate the role of $TGF-{\beta}$ downregulation in cancer cell death. We found that the downregulation of $TGF-{\beta}$ increased the phosphorylation of several SAPKs, such as p38 and JNK. Moreover, reactive oxygen species (ROS) production was also increased by $TGF-{\beta}$ downregulation, which triggered Akt inactivation and NOX4 increase-derived ROS in a cancer cell-type-specific manner. We also revealed the possibility of substantial gene fluctuation in response to $TGF-{\beta}$ downregulation related to SAPKs. The expression levels of Trx and GSTM1, which encode inhibitory proteins that bind to ASK1, were reduced, likely a result of the altered translocation of Smad complex proteins rather than from ROS production. Instead, both ROS and ROS-mediated ER stress were responsible for the decrease in interactions between ASK1 and Trx or GSTM1. Through these pathways, ASK1 was activated and induced cytotoxic tumor cell death via p38/JNK activation and (or) induction of ER stress.

• Tuberculosis and Respiratory Diseases
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• v.58 no.1
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• pp.18-24
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• 2005

Background : Endobronchial tuberculosis often complicates bronchostenosis, which can cause dyspnea due to an airway obstruction, and can be misdiagnosed as bronchial asthma or lung cancer. This study investigated the possible correlation between the $interferon-{\gamma}$($IFN-{\gamma}$) and transforming growth $factor-{\beta}$($TGF-{\beta}$) levels in the serum and bronchial washing fluid and the treatment results in endobronchial tuberculosis patients. Methods : Sixteen patients, who were diagnosed as endobronchial tuberculosis using bronchoscopy, and 10 healthy control subjects were enrolled in this study. The $IFN-{\gamma}$ and $TGF-{\beta}$ levels were measured in the serum and bronchial washing fluid of 16 endobronchial tuberculosis patients before and after treatment using the ELISA method. The endobronchial tuberculosis patients were divided into those who showed bronchial fibrostenosis after treatment and those who did not. Results : The $IFN-{\gamma}$ and $TGF-{\beta}$ levels in the bronchial washing fluid in endobronchial tuberculosis patients were elevated comparing to the control (p<0.05). After treatment, 7 of the 16 endobronchial tuberculosis patients showed bronchial fibrostenosis and the other 9 cases healed without this sequela. In the patients with fibrostenosis after treatment, the initial serum $TGF-{\beta}$ level was lower than the patients without fibrostenosis after treatment (p<0.05). Moreover, the serum $TGF-{\beta}$ level after treatment further decreased comparing to the patients without fibrostenosis after treatment(p<0.05). Conclusion : Elevated $IFN-{\gamma}$ and $TGF-{\beta}$ levels in the bronchial washing fluid in endobronchial tuberculosis patients are believed to be related to the pathogenesis of endobronchial tuberculosis. The decreased initial serum $TGF-{\beta}$ level and the change in the serum $TGF-{\beta}$ level after treatment are believed to be involved in bronchial fibrostenosis during the course of the disease.

• BMB Reports
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• v.50 no.8
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• pp.429-434
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• 2017

Endometriosis is the abnormal growth of endometrial cells outside the uterus, causing pelvic pain and infertility. Furthermore, adhesion of endometrial tissue fragments to pelvic mesothelium is required for the initial step of endometriosis formation outside uterus. $TGF-{\beta}1$ and adhesion molecules importantly function for adhesion of endometrial tissue fragments to mesothelium outside uterus. However, the function of $TGF-{\beta}1$ on the regulation of adhesion molecule expression for adhesion of endometrial tissue fragments to mesothelium has not been fully elucidated. Interestingly, transforming growth factor ${\beta}1$ ($TGF-{\beta}1$) expression was higher in endometriotic epithelial cells than in normal endometrial cells. The adhesion efficiency of endometriotic epithelial cells to mesothelial cells was also higher than that of normal endometrial cells. Moreover, $TGF-{\beta}1$ directly induced the adhesion of endometrial cells to mesothelial cells through the regulation of integrin of ${\alpha}V$, ${\alpha}6$, ${\beta}1$, and ${\beta}4$ via the activation of the $TGF-{\beta}1/TGF-{\beta}RI/Smad2$ signaling pathway. Conversely, the adhesion of $TGF-{\beta}1-stimulated$ endometrial cells to mesothelial cells was clearly reduced following treatment with neutralizing antibodies against specific $TGF-{\beta}1-mediated$ integrins ${\alpha}V$, ${\beta}1$, and ${\beta}4$ on the endometrial cell membrane. Taken together, these results suggest that $TGF-{\beta}1$ may act to promote the initiation of endometriosis by enhancing integrin-mediated cell-cell adhesion.

• The korean journal of orthodontics
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• v.28 no.4 s.69
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• pp.627-640
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• 1998

[$TGF-{\beta}$] is a polypeptide with multiple physiological functions in regulation of cell-to-cell interaction and in growth and development. The active form of vitmain $D_3$, 1,25-dihydroxycholecalciferol $[1,25-(OH)_2D_3]$, is one of the most potent stimulators of osteoclastic acitivity. The purpose of this study was to evaluate the effect of Vitamin $D_3$ and/or $TGF-{\beta}$ on the periodontal ligament(PDL) cells. Human PDL cells were prepared from the first premolars extracted for the orthodontic treatment and were incubated in the environment of , $37^{\circ},\;5\%\;CO_2\;and\;95\%$ humidity. 10, 50 or 100ng/m1 of $1,25-(OH)_2D_3$ and 0.1, 1, 5 or 10ng/ml of $TGF-{\beta}$ were administered to the culture wells, separately or in combination. And the viability of PDL cells was evaluated by MTT assay The obtained results were as follows. 1. The viability of PDL cells in 10ng/ml of vitamin $D_3$ was not significantly differenent from that of the control group at 1, 2 and 3-day of cultivation, but it was significantly increased in 50ng/ml of Vitamin $D_3$ at 3-day and in 100ng/m1 of Vitamin $D_3$ at 2 and 3-day. 2. The viability of PDL cells in 0.1ng/ml of $TGF-{\beta}$ was not significantly differenent from that of the control group at 1, 2 and 3-day of cultivation, but it was significantly increased in 1 and 5ng/ml of $TGF-{\beta}$ at 3-day of cultivation, and in 10ng/ml of $TGF-{\beta}$ at 2 and 3-day of cultivation. 3. In case of admixture of 1ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$, the viability of PDL cells was significantly increased in the admixture of 100ng/ml of vitamin $D_3$ at 3-day of cultivation 4. In case of admixture of 5ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$, the viability of PDL cells began to be increased from 2-day of cultivation in the admixture of 10 50 and 100ng/ml of vitamin $D_3$, but it was not maintained at 3-day in the admixture of 10ng/m of vitamin $D_3$. 5. In case of admixture of 10ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$ the viability of PDL cells was significantly increased in the admixture of 50ng/ml of vitamin $D_3$ at 2 and 3-day of cultivation, and in the admixture of 100ng/ml at 1, 2 and 3-day.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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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.

• Proceedings of the Korean Society of Toxicology Conference
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• 2003.10b
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• pp.142-142
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• 2003

Transforming growth factor (TGF)-${\beta}$, a hormonally active polypeptide found in normal and transformed tissue, is a potent regulator of cell growth and differentiation. In this study, we examined the effect of TGF-${\beta}$ on invasion and motility of MCF10A human breast epithelial cells. TGF-${\beta}$ induced migration and invasive phenotype of the parental MCF10A cells in a dose-dependent manner.(omitted)

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.152.2-153
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• 2003

${\beta}Although$ it has been demonstrated that $p2l^{WAFl/Cip1}$, a well known cell cycle inhibitor, could be induced by $TGF-{\beta}$ in a p53-independent manner, the detailed signal transduction pathways still remain poorly understood. In this study, we show that ERK is required for $TGF-{\beta}$ induction of $p21^{WAF1/Cip1}$, but JNK or p38 MAPK is not. ERK activation by $TGF-{\beta}$ significantly attenuated by treatment with ROS scavenger such as NAC or catalase, indicating that ROS, mainly $H_2O_2$, generation by $TGF-{\beta}$ might stimulate ERK signaling pathway to require the induction of $p21^{WAF1/Cip1}$. (omitted)

• Archives of Pharmacal Research
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• v.25 no.6
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• pp.903-909
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• 2002

Research in the cytokine field has grown exponentially in recent years, and the validity of such studies relies heavily on the appropriate measurement of levels of cytokines in various biological samples. Transforming growth factor (TGF)-$\beta$, a hormonally active polypeptide found in normal and transformed tissue, is a potent regulator of cell growth and differentiation. The most widely used bioassay for TGF-$\beta$ is the inhibition of the proliferation of mink lung epithelial cells. Though detection of [$^3$H]thymidine incorporation is more sensitive than the MTT assay, it presents some disadvantages due to the safety and disposal problems associated with radioisotopes. In this study, we attempted to ascertain the experimental conditions which could be used for measuring the in vitro biological activity of TGF-$\beta$ in a safer and more sensitive way compared with the currently available methods. We compared the commonly used method, the MTT assay, to the XTT assay using different parameters including cell number, incubation time and the wave length used for detecting the product. We examined the anti-proliferative activities of TGF-$\beta$ in three different cell lines: Mv-1-Lu mink lung epithelial cells, MCF10A human breast epithelial cells and H-ras-transformed MCF10A cells. Herein, we present an experimental protocol which provides the most sensitive method of quantifying the biological activity of TGF-$\beta$, with a detection limit of as low as 10 pg/ml: Mv-1-Lu or H-ras MCF10A cells ($1{\times}10^5/well$) were incubated with TGF-$\beta$ at $37^{\circ}C$ in a humidified $CO_2$ incubator for 24 hr followed by XTT treatment and determination of absorbance at 450 or 490 nm. Our results may contribute to the establishment of an in vitro bioassay system, which could be used for the satisfactory quantitation of TGF-$\beta$.

• The Korea Journal of Herbology
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• v.28 no.4
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• pp.49-55
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• 2013

Objectives : Mori Folium was popularly used as one of the traditional medicinal herbs. Although M. Folium has been cultivated for rearing silkworm historically, it's use has been expanded as natural therapeutic agent for the treatment of filariasis, diabetes and dropsy in East Asia. However, little has been known about the effect of M. Folium on liver fibrosis. Therefore, we would like to explore an anti-fibrogenic potential of M. Folium extract (MFE) using immortalized human hepatic stellate cell line, LX-2 cells. Methods : We examined the effects of MFE on the transforming growth factor ${\beta}1$ ($TGF{\beta}1$)-induced liver fibrosis in LX-2 cells. Cell viability, Smad binding element-driven luciferase activity, phosphorylations level of Smad 2/3, and expression level of $TGF{\beta}1$-dependent target genes were monitored in the MFE-treated LX-2 cells. Results : Up to 30 ${\mu}g/ml$ MFE treatment did not show any possible toxic effect in LX-2 cells. MFE inhibited $TGF{\beta}1$-inducible Smad binding element-driven luciferase activity and decreased the $TGF{\beta}1$-inducible phosphorylations of Smad 2 and Smad 3 in hepatic stellate cell in a dose dependent manner. Furthermore, increases of plasminogen activator inhibitor type 1, $TGF{\beta}1$ and matrix metalloproteinases 2 genes by $TGF{\beta}1$ were also attenuated by MFE treatment. Conclusions : These findings suggested that MFE would be used as a potential therapeutic agent for the treatment liver fibrosis, which might be mediated by the inhibition of $TGF{\beta}1$-inducible Smad 2/3 transactivation and target genes expression.

• Reproductive and Developmental Biology
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• v.30 no.4
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• pp.271-277
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• 2006

Transforming growth $factor-{\beta}\;(TGF-{\beta})$ has been shown to have a positive effect on in vitro fertilization (IVF) and has been reported to stimulate meiosis at follicular level in variety of species. The study was designed to determine the expression patterns of $TGF-{\beta}1,\;TGF-{\beta}$ receptors type I, II and Smads gene in bovine oocytes and embryos. $TGF-{\beta}1$ and their receptors were observed in the unfertilized oocytes. $TGF-{\beta}1$ and type II receptor were not expressed at the blastocyst stage, however, only type I receptor was exclusively observed at the same stage. The blastocyst stage, in particular, showed high levels of mRNA expression patterns containing a $TGF-{\beta}1$ type I receptor. The mRNA expression pattern of Smad 2 at all stages of embryonic development was similar in all respect with $TGF-{\beta}1$ type I receptor. On the contrary, Smad 3 and 4 were expressed with high and low level mRNA at the blastocyst stage. In conclusion. it is suggested that $TGF-{\beta}1$ signaling may be regarded as an important entity during the preimplantation embryo development.