• Molecules and Cells
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• 제23권1호
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• pp.23-29
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• 2007

Cyclic AMP-responsive element binding protein (CREB) is known to be associated with angiogenesis. In the present study we investigated the possible role of CREB in the expression of vascular endothelial growth factor (VEGF) by mouse macrophages. Over-expression of CREB increased VEGF secretion by cells of the RAW264.7 mouse macrophage cell line. It also increased the promoter activity of a mouse reporter driven by the VEGF promoter, while a dominant negative CREB (DN-CREB) abrogated the activity, suggesting that CREB mediates VEGF transcription. Forskolin, an adenylyl cyclase activator, stimulated VEGF transcription, and the PKA inhibitor H89 abolished this effect. IFN-${\gamma}$, a potent cytokine, stimulated VEGF expression only in part through the PKA-CREB pathway. These results indicate that PKA phosphorylates CREB and so induces VEGF gene expression. An analysis of mutant promoters revealed that one of the putative CREB responsive elements (CREs), at -399 ~ -388 in the promoter, is critical for CREB-mediated VEGF promoter activity, and the significance of this CRE was confirmed by chromatin immunoprecipitation assays.

• BMB Reports
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• 제51권6호
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• pp.296-301
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• 2018

Mitochondrial DNA (mtDNA) mutations are often observed in various cancer types. Although the correlation between mitochondrial dysfunction and cancer malignancy has been demonstrated by several studies, further research is required to elucidate the molecular mechanisms underlying accelerated tumor development and progression due to mitochondrial mutations. We generated an mtDNA-depleted cell line, ${\rho}^0$, via long-term ethidium bromide treatment to define the molecular mechanisms of tumor malignancy induced by mitochondrial dysfunction. Mitochondrial dysfunction in ${\rho}^0$ cells reduced drug-induced cell death and decreased the expression of pro-apoptotic proteins including p53. The p53 expression was reduced by activation of nuclear $factor-{\kappa}B$ that depended on elevated levels of free calcium in $HCT116/{\rho}^0$ cells. Overall, these data provide a novel mechanism for tumor development and drug resistance due to mitochondrial dysfunction.

• Molecular & Cellular Toxicology
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• 제2권2호
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• pp.97-105
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• 2006

Several features of the implant surface, such as roughness, topography, and composition play a relevant role in implant integration with bone. This study was conducted in order to determine the effects of ceramic-coatings on Ti surfaces on the biological responses of a human osteoblast-like cell line (MG63). MG63 cells were cultured on Zr (Zrconium-coated surface), Nb (Niobium-coated surface), and control (Uncoated Titanium) Ti. The morphology of these cells was assessed by SEM. The cDNAs prepared from the total RNAs of the MG63 were hybridized into a human cDNA microarray (1,152 elements). The appearances of the surfaces observed by SEM were different on each of the three dental substrate types. MG63 cells cultured on Zr, Nb and control exhibited cell-matrix interactions. In the expression of several genes were up-, and down-regulated on the different surfaces. The attachment and expression of key osteogenic regulatory genes were enhanced by the surface morphology of the dental materials used.

• Clinical and Experimental Pediatrics
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• 제64권7호
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• pp.355-363
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• 2021

Background: Nephrotic syndrome (NS) is a common renal disorder in children attributed to podocyte injury. However, children with the same diagnosis have markedly variable treatment responses, clinical courses, and outcomes, suggesting molecular heterogeneity. Purpose: This study aimed to explore the molecular responses of podocytes to nephrotic plasma to identify specific genes and signaling pathways differentiating various clinical NS groups as well as biological processes that drive injury in normal podocytes. Methods: Transcriptome profiles from immortalized human podocyte cell line exposed to the plasma of 8 subjects (steroid-sensitive nephrotic syndrome [SSNS], n=4; steroid-resistant nephrotic syndrome [SRNS], n=2; and healthy adult individuals [control], n=2) were generated using microarray analysis. Results: Unsupervised hierarchical clustering of global gene expression data was broadly correlated with the clinical classification of NS. Differential gene expression (DGE) analysis of diseased groups (SSNS or SRNS) versus healthy controls identified 105 genes (58 up-regulated, 47 down-regulated) in SSNS and 139 genes (78 up-regulated, 61 down-regulated) in SRNS with 55 common to SSNS and SRNS, while the rest were unique (50 in SSNS, 84 genes in SRNS). Pathway analysis of the significant (P≤0.05, -1≤ log2 FC ≥1) differentially expressed genes identified the transforming growth factor-β and Janus kinase-signal transducer and activator of transcription pathways to be involved in both SSNS and SRNS. DGE analysis of SSNS versus SRNS identified 2,350 genes with values of P≤0.05, and a heatmap of corresponding expression values of these genes in each subject showed clear differences in SSNS and SRNS. Conclusion: Our study observations indicate that, although podocyte injury follows similar pathways in different clinical subgroups, the pathways are modulated differently as evidenced by the heatmap. Such transcriptome profiling with a larger cohort can stratify patients into intrinsic subtypes and provide insight into the molecular mechanisms of podocyte injury.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5659-5663
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• 2012

To investigate the expression of hPOT1 in the HeLa cell line and screen point mutations of hpot1 in different tumor tissues a two step osmotic method was used to extract nuclear proteins. EMSA was performed to determine the expression of hPOT1 in the HeLa cell line. PCR was also employed to amplify the exon14 sequence of the hpot1 gene in various of cancer tissues. A SV gel and PCR clean-up system was performed to enrich PCR products. DNAStar was used to analyse the exon14 sequence of the hpot1 gene. hPOT1 was expressed in the HeLa cell line and the signal was gradually enhanced as the amount of extracted nuclear proteins increased. The DNA fragment of exon14 of hpot1 was successfully amplified in the HeLa cell line and all cancer tissues, point mutations being observed in 2 out of 3 cases of endometrial cancer (66.7%) despite the hpot1 sequence being highly conserved. However, the sequence of hpot1 exon14 do not demonstrate point mutations in most cancer tissues. Since hPOT1 was expressed in HeLa cell and the probability of gene point variants was obviously higher in endometrial cancer than other cancers, it may be involved in the pathogenesis of gynecological cancers, especially in cervix and endometrium.

• 천문학회보
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• 제38권2호
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• pp.65.2-65.2
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• 2013

We present a study of outflows on 24 embedded young stellar objects (YSOs), which are selected from the sources of the Dust, Ice, and Gas in Time (DIGIT) Herschel key program. Molecular outflow activity, which is believed to have strong dependence on accretion process, is the most powerful in the early embedded phase of star formation and declines as the central protostars evolve to the main sequence stage. In order to study the relation between the CO outflow observed in low J transitions and the properties of protostars, we mapped the CO outows of the selected targets in J = 1-0 and J = 2-1 lines with the 14-m TRAO telescope and the 6-m SRAO telescope, respectively. We estimate CO outflow momentum fluxes (Fco) and compare with bolometric luminosity, Lbol, bolometric temperature, Tbol, and the FIR molecular line luminosities of CO, $H_2O$, OH and [O I], which were detected by the Herschel-PACS observations. We found that $Fco_{1-0}$ is greater than $Fco_{2-1}$, and the mean ratio is about 2. L1455-IRS3 and IRAM04191 have high Fco in spite of low $L_{bol}$. The well known correlation between Fco and $L_{bol}$. is not very evident from all our samples. However, Fco and $L_{bol}$. show a rather strong correlation if L1455-IRS3 and IRAM04191 are excluded. Fco shows little correlation with FIR line luminosities of individual species, while the total FIR line luminosity summed over CO, $H_2O$, OH, and [OI] lines seems to have some correlation. In addition, we report 22 GHz $H_2O$, and 44 GHz CH3OH maser line detections in four sources out of the 24 YSOs.

• 천문학회보
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• 제39권1호
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• pp.63.2-63.2
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• 2014

As a part of the "Dust, Ice, and Gas In Time" (DIGIT) key program on Herschel, we observed GSS30-IRS1, a Class I protostar located in Ophiuchus (d =125 pc), with Herschel/Photodetector Array Camera and Spectrometer (PACS). More than 70 lines were detected within a wavelength range from 50 ${\mu}m$ to 200 ${\mu}m$: CO lines from J = 14-13 to 41-40, several $H_2O$ lines of Eup = 100 K to 1500 K, 16 transitions of OH rotational lines, and two atomic [O I] lines at 63 and 145 ${\mu}m$. The [C II] line, known as a tracer of externally heated gas by the interstellar radiation field, is also detected at 158 ${\mu}m$. All lines, except [O I] and [C II], are detected only at the central spaxel of $9^{\prime\prime}.4{\times}9^{\prime\prime}.4$. The [O I] emission is extended along a NE-SW orientation, which is consistent with the known outflow direction, while the [C II] line is detected over all spaxels. One possible explanation of the detection of the [C II] line and no correlation of its spatial distribution with any other molecular emission is the existence of the enhanced ISRF nearby GSS30-IRS1. One interesting feature of GSS30-IRS1 is that the continuum emission is extended beyond the point-spread function (PSF), unlike the molecular line emission, indicative of significant external heating. The best-fit continuum model of GSS30-IRS1 with the physical structure including flared disk, envelope, and outflow shows that the internal luminosity is 11 $L_{\odot}$, and the region is also externally heated by a radiation field enhanced by a factor of 25 compared to the local standard interstellar field.

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권1호
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• pp.1-5
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• 1998

One cervical cancer cell line, C9, carrying human papillomavirus type 18 (HPV18) genes that is one of the major etiologic concoviruses for cervical cancer was characterized. This cell line was further characterized for its capacity related to the epithelial cell proliferation, stratification and differentiation in reconstituted artificial epithelial tissue. The in vitro construction of three dimensional artificial cervical opithelial tissue has been engineered using C9 epithelial cancer cells, human foreskin fibroblasts and a matrix made of type I collagen by organotypic culture of epithelial cells. The morphology of paraffin embedded artificial tissue was examined by histochemical staining. The artificial epithelial tissues were well developed having multilayer. However, the tissue morphology was similar to the cervical tissus having displasia induced by HPV infection. The characteristics of the artificial tissues were examined by determinining the expression of specific marker proteins. In the C9 derived artificial tissues, the expression of EGF receptor, as epithelial proliferation marker proteins for stratum basale was observed up to the stratum spinosum. Another epithelial proliferation marker for stratum spinosum, cytokerations 5/6/18, were observed well over the stratum spinosum. For the differentiation markers, the expression of involucrin and filaggrin were observed while the terminal differentiation marker, cytokeratins 10/13 was not detected at all. Therefore the reconstituted artificial epithelial tissues expressed the same types of differentiation marker proteins that are expressed in normal human cervical epithelial tissues but lacked the final differentiation capacity representing characteristics of C9 cell line as a cancer tissue devived cell line. Expression of HPV18 E6 oncoprotein was also observed in this artifical cervical opithelial tissue though the intensity of the staining was weak. Thus this artificial epithelial tissue could be used as a useful model system to examine the relationship between HPV-induced cervical oncogenesis and epithelial cell differentiation.

• Journal of Microbiology and Biotechnology
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• 제10권3호
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• pp.287-292
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• 2000

Recombinant Aspergillus niger was constructed to express and secrete a biologically active murine granulaocyte macrophage-colony stimulating factor (mGM-CSF). A 500 bp fragment encoding the signal peptide and terminator of glyceraldehyde-3-phosphate dehydrogenase (gpd). The hygromycin phosphotrasferase gene (hph) was used as a selection marker for the fungal transformants. An expression vector was introduced into A. niger ATCC 9642, and a Northern blot analysis indicated the presence of a considerable amount of transcripts from the introduced mGM-CSF. The biological activity of recombinant mGM-CSF (rmGM-CSF) isolated from the culture filtrate was confirmend by measuring the proliferationof the GM-CSF dependent FDC-P1 cell line. It appeared that rmGM-CSF was amenable to the proteolytic activity produced by A. niger, since biological actibity was only observed when the transformants were grown in a protease-repressing medium, and the activity of rmGM-CSF dramatically decreased with an increase of age of the culture. The yield of rmGM-CSF, as determined by ELISA. was 640 ng/l of culture filtrate. Accordingly, its specific activity is estimated to be approximately two-and-a-half times higher than that of a commercial preparation from E. coli.

• 천문학회지
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• 제34권3호
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• pp.157-166
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• 2001

The molecular cloud associated with the H II region S301 has been mapped in the J = 1-0 transitions of $^{12}CO$ and $^{13}CO$ using the 13.7 m radio telescope of Taeduk Radio Astronomy Observatory. The cloud is elongated along the north-south direction with two strong emission components facing the H II region. Its total mass is $8.7 {\times} 10^3 M{\bigodot}$. We find a velocity gradient of the molecular gas near the interface with the optical H II region, which may be a signature of interaction between the molecular cloud and the H II region. Spectra of CO, CS, and HCO+ exhibit line splitting even in the densest part of the cloud and suggests the clumpy structure. The radio continuum maps show that the ionzed gas is distributed with some asymmetry and the eastern part of the H II region is obscured by the molecular cloud. We propose that the S301 H II region is at the late stage of the champagne phase, but the second generation of stars has not yet been formed in the postshock layer.