• Title/Summary/Keyword: Reporter

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Ets-1 enhances tumor migration through regulation of CCR7 expression

  • Fang, Li-Wen;Kao, Ying-Hsien;Chuang, Ya-Ting;Huang, Huey-Lan;Tai, Tzong-Shyuan
    • BMB Reports
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    • v.52 no.9
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    • pp.548-553
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    • 2019
  • Ets-1 is a prototype of the ETS protein family. Members of the ETS protein family contain a unique ETS domain. Ets-1 is associated with cancer progression and metastasis in many types of cancer. Many studies have shown a link between elevated expression of Ets-1 in cancer biopsies and poor survival. CCR7 is a chemokine that binds to specific ligand CCL21/CCL19. CCR7 expression is associated with tumor metastasis and infiltration into lymph nodes. The objective of this study was to test whether Ets-1 could regulate CCR7 expression and enhance tumor metastasis. Our data showed that CCR7 expression was downregulated in Ets-1-deficient T cells upon T-cell stimulation. Overexpression of Ets-1 increased CCR7 expression in breast cancer cell lines. In contrast, knockdown of Ets-1 reduced CCR7 expression. Ets-1 could directly bind to CCR7 promoter and mediate CCR7 expression in luciferase reporter assays and chromatin immunoprecipitation assays. Transactivation activity of Ets-1 was independent of the Pointed domain of Ets-1. Ets-1 could also enhance $NF-{\kappa}B$ and CBP transactivation of CCR7 promoter. Our results also showed that Ets-1 could modulate cancer cell transmigration by altering CCR7 expression in transwell assay and wound healing assay. Taken together, our data suggest that Ets-1 can enhance CCR7 expression and contribute to tumor cell migration.

Antioxidant and Anticancer Effects of Water Extract from Pleurotus eryngii, Flammulina velutipes (새송이버섯, 팽이버섯 열수추출물의 항산화 및 항암 활성)

  • Ryu, Hye Sook;Kim, Soo Hyun;Jeon, Moon Hee;Choi, Hae Yeon
    • The Korean Journal of Food And Nutrition
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    • v.31 no.6
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    • pp.911-918
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    • 2018
  • Various studies on the effect of mushroom, a member of phytonutrients, on inflammatory diseases have been reporter over the years, Among various species mushrooms, King oyster mushroom (Pleurotus eryngii) and winter mushroom (Flammulina velutipes) are popular dietary ingredients for Asian cuisine. The extracts from all the three mushrooms were used to determine the antioxidative effect and the cytotoxicity. Result analysis were repeated more than three times to get an average${\pm}$standard deviation, and statistical significance were confirmed by SPSS. As a result, total phenol content of the king oyster mushroom and the winter mushroom were $19.66{\pm}0.10mg/g$ and $22.08{\pm}1.10mg/g$, respectively. Also, the total flavonoid content was $15.21{\pm}1.31mg/g$ for the king oyster mushroom and $20.50{\pm}4.52mg/g$ for winter mushroom. The results in winter mushroom showed higher values in total phenol and flavonoid content than in the king oyster mushroom. All samples of extracts showed free radical DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activity with their 10 mg/mL concentration. These results indicate that the extract of the king oyster mushroom (Pleurotus eryngii) and the winter mushroom (Flammulina velutipes) may be possible phytonutrients with the anti-inflammatory and the anti-cancer effect.

Drosophila CrebB is a Substrate of the Nonsense-Mediated mRNA Decay Pathway that Sustains Circadian Behaviors

  • Ri, Hwajung;Lee, Jongbin;Sonn, Jun Young;Yoo, Eunseok;Lim, Chunghun;Choe, Joonho
    • Molecules and Cells
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    • v.42 no.4
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    • pp.301-312
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    • 2019
  • Post-transcriptional regulation underlies the circadian control of gene expression and animal behaviors. However, the role of mRNA surveillance via the nonsense-mediated mRNA decay (NMD) pathway in circadian rhythms remains elusive. Here, we report that Drosophila NMD pathway acts in a subset of circadian pacemaker neurons to maintain robust 24 h rhythms of free-running locomotor activity. RNA interference-mediated depletion of key NMD factors in timeless-expressing clock cells decreased the amplitude of circadian locomotor behaviors. Transgenic manipulation of the NMD pathway in clock neurons expressing a neuropeptide PIGMENT-DISPERSING FACTOR (PDF) was sufficient to dampen or lengthen free-running locomotor rhythms. Confocal imaging of a transgenic NMD reporter revealed that arrhythmic Clock mutants exhibited stronger NMD activity in PDF-expressing neurons than wild-type. We further found that hypomorphic mutations in Suppressor with morphogenetic effect on genitalia 5 (Smg5) or Smg6 impaired circadian behaviors. These NMD mutants normally developed PDF-expressing clock neurons and displayed daily oscillations in the transcript levels of core clock genes. By contrast, the loss of Smg5 or Smg6 function affected the relative transcript levels of cAMP response element-binding protein B (CrebB) in an isoform-specific manner. Moreover, the overexpression of a transcriptional repressor form of CrebB rescued free-running locomotor rhythms in Smg5-depleted flies. These data demonstrate that CrebB is a rate-limiting substrate of the genetic NMD pathway important for the behavioral output of circadian clocks in Drosophila.

LINC01272 Suppressed Cell Multiplication and Induced Apoptosis Via Regulating MiR-7-5p/CRLS1 Axis in Lung Cancer

  • Ma, Xuan;Liu, Yang;Tian, Hao;Zhang, Bo;Wang, Meiling;Gao, Xia
    • Journal of Microbiology and Biotechnology
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    • v.31 no.7
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    • pp.921-932
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    • 2021
  • LINC01272 is a long non-coding RNA (lncRNA) that has been considered as a biomarker for many diseases including lung squamous cell carcinoma. Here, we investigated the function and mechanism of LINC01272 on lung cancer (LC). The differential expression of LINC01272 in LC and normal samples was analyzed by GEPIA based on the data from TCGA-LUAD database, as survival prognosis was analyzed through Kaplan-Meier Plotter. LINC01272 overexpression plasmid and miR-7-5p mimic were transfected into A549 and PC-9 cells. LINC01272, miR-7-5p and cardiolipin synthase 1 (CRLS1) mRNA expression was measured by quantitative reverse transcription-polymerase chain reaction. Cell viability was detected through MTT assay. Cell multiplication was evaluated by cell formation assay. Cell apoptosis was assessed through flow cytometry assay. Through bioinformatics, the target miRNA of LINC01272 and downstream genes of miR-7-5p were predicted. The targeting relationship was tested by dual luciferase reporter analysis. CRLS1, B-cell lymphoma-2 (Bcl-2), BCL2-associated X (Bax) and cleaved caspase-3 protein levels were detected through western blot. LINC01272 was downregulated in LC and low LINC01272 expression had poor prognosis. In A549 and PC-9 cells, LINC01272 inhibited cell viability and multiplication and induced apoptosis. LINC01272 negatively regulated miR-7-5p and CRLS1 was a target of miR-7-5p. MiR-7-5p reversed the effect of LINC01272 on viability, multiplication, apoptosis and expression of miR-7-5p and CRLS1 as well as apoptosis-related factors (Bcl-2, Bax and cleaved caspase-3). LINC01272 suppressed cell multiplication and induced apoptosis via regulating the miR-7-5p/CRLS1 axis in LC.

Identification and Validation of Four Novel Promoters for Gene Engineering with Broad Suitability across Species

  • Wang, Cai-Yun;Liu, Li-Cheng;Wu, Ying-Cai;Zhang, Yi-Xuan
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1154-1162
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    • 2021
  • The transcriptional capacities of target genes are strongly influenced by promoters, whereas few studies have focused on the development of robust, high-performance and cross-species promoters for wide application in different bacteria. In this work, four novel promoters (Pk.rtufB, Pk.r1, Pk.r2, and Pk.r3) were predicted from Ketogulonicigenium robustum and their inconsistency in the -10 and -35 region nucleotide sequences indicated they were different promoters. Their activities were evaluated by using green fluorescent protein (gfp) as a reporter in different species of bacteria, including K. vulgare SPU B805, Pseudomonas putida KT2440, Paracoccus denitrificans PD1222, Bacillus licheniformis and Raoultella ornithinolytica, due to their importance in metabolic engineering. Our results showed that the four promoters had different activities, with Pk.r1 showing the strongest activity in almost all of the experimental bacteria. By comparison with the commonly used promoters of E. coli (tufB, lac, lacUV5), K. vulgare (Psdh, Psndh) and P. putida KT2440 (JE111411), the four promoters showed significant differences due to only 12.62% nucleotide similarities, and relatively higher ability in regulating target gene expression. Further validation experiments confirmed their ability in initiating the target minCD cassette because of the shape changes under the promoter regulation. The overexpression of sorbose dehydrogenase and cytochrome c551 by Pk.r1 and Pk.r2 resulted in a 22.75% enhancement of 2-KGA yield, indicating their potential for practical application in metabolic engineering. This study demonstrates an example of applying bioinformatics to find new biological components for gene operation and provides four novel promoters with broad suitability, which enriches the usable range of promoters to realize accurate regulation in different genetic backgrounds.

KAT8/MOF-Mediated Anti-Cancer Mechanism of Gemcitabine in Human Bladder Cancer Cells

  • Zhu, Huihui;Wang, Yong;Wei, Tao;Zhao, Xiaoming;Li, Fuqiang;Li, Yana;Wang, Fei;Cai, Yong;Jin, Jingji
    • Biomolecules & Therapeutics
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    • v.29 no.2
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    • pp.184-194
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    • 2021
  • Histone acetylation is a well-characterized epigenetic modification controlled by histone acetyltransferases (HATs) and histone deacetylases (HDACs). Imbalanced histone acetylation has been observed in many primary cancers. Therefore, efforts have been made to find drugs or small molecules such as HDAC inhibitors that can revert acetylation levels to normal in cancer cells. We observed dose-dependent reduction in the endogenous and exogenous protein expression levels of KAT8 (also known as human MOF), a member of the MYST family of HATs, and its corresponding histone acetylation at H4K5, H4K8, and H4K16 in chemotherapy drug gemcitabine (GEM)-exposed T24 bladder cancer (BLCA) cells. Interestingly, the reduction in MOF and histone H4 acetylation was inversely proportional to GEM-induced γH2AX, an indicator of chemotherapy drug effectiveness. Furthermore, pGL4-MOF-Luc reporter activities were significantly inhibited by GEM, thereby suggesting that GEM utilizes an MOF-mediated anti-BLCA mechanism of action. In the CCK-8, wound healing assays and Transwell® experiments, the additive effects on cell proliferation and migration were observed in the presence of exogenous MOF and GEM. In addition, the promoted cell sensitivity to GEM by exogenous MOF in BLCA cells was confirmed using an Annexin V-FITC/PI assay. Taken together, our results provide the theoretical basis for elucidating the anti-BLCA mechanism of GEM.

AI Advisor for Response of Disaster Safety in Risk Society (위험사회 재난 안전 분야 대응을 위한 AI 조력자)

  • Lee, Yong-Hak;Kang, Yunhee;Lee, Min-Ho;Park, Seong-Ho;Kang, Myung-Ju
    • Journal of Platform Technology
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    • v.8 no.3
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    • pp.22-29
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    • 2020
  • The 4th industrial revolution is progressing by country as a mega trend that leads various technological convergence directions in the social and economic fields from the initial simple manufacturing innovation. The epidemic of infectious diseases such as COVID-19 is shifting digital-centered non-face-to-face business from economic operation, and the use of AI and big data technology for personalized services is essential to spread online. In this paper, we analyze cases focusing on the application of artificial intelligence technology, which is a key technology for the effective implementation of the digital new deal promoted by the government, as well as the major technological characteristics of the 4th industrial revolution and describe the use cases in the field of disaster response. As a disaster response use case, AI assistants suggest appropriate countermeasures according to the status of the reporter in an emergency call. To this end, AI assistants provide speech recognition data-based analysis and disaster classification of converted text for adaptive response.

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Development of sandwich enzyme-linked immunosorbent assay for a large-scale detection of porcine transmissible gastroenteritis virus in feces

  • Oh, Yeonsu;Lee, Sang-Joon;Cho, Ho-Seong;Tark, Dongseob
    • Korean Journal of Veterinary Service
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    • v.43 no.4
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    • pp.237-244
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    • 2020
  • Porcine transmissible gastroenteritis (TGE) has been a significant cause of economic losses in pig farming industry since 1950s. Although transmissible gastroenteritis virus (TGEV) has declined in recent years, it should not be excluded because of its characteristics; the frequency of gene mutation, the mortality in piglets, and the possibility for sudden incidence. Therefore, the herd-level monitoring of the virus is important to prevent further circulation of TGE. The aim of this study is to develop a large-scale sandwich enzyme-linked immunosorbent assay (ELISA) with high specificity to rapidly detect TGEV in feces by using monoclonal antibodies (Mabs). The TGEV specific Mabs were produced in hybridoma cells. Among the Mabs belonged to the IgG class developed by this study, the final selected 8H6, 1B7, 4G3, and 1F8 were identified to have the neutralization ability against TGEV. The sandwich ELISA was established using 8H6 as a reporter antibody and 1B7 and the reported 5C8 as a capture antibody. The developed sandwich ELISA was able to distinguish TGEV from other pathogenic diarrheal agents (porcine rotavirus, porcine reovirus, porcine epidemic diarrhea virus (PEDV), E. coli, and C. perfringens) in tissue culture as well as fecal samples. And the detection rate of TGEV in feces was 80% compared with RT-PCR. The results suggested that the developed sandwich ELISA may be useful in the herd-level monitoring for effective preventive measures due to the early diagnosis of TGEV using a large amount of samples.

How "Covid-19" Affected Reporters' News Coverage?: Focusing on Reporters' Perception of Changes in Work Environment Before and After the Pandemic (코로나 19는 기자들의 취재관행에 어떤 영향을 주었나?: 팬데믹 전후의 근무형태 변화에 대한 기자 인식을 중심으로)

  • Yang, Young-Yu
    • The Journal of the Korea Contents Association
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    • v.21 no.5
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    • pp.11-21
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    • 2021
  • The purpose of this study is to explore and analyze how the Covid-19 pandemic has affected the reporting practices and news production of the reporters working with Korean media over the past one year. To this end, this study has conducted in-depth interviews with reporters working with daily newspapers, news agencies, and broadcasting companies. The analysis of the interviews resulted in the following generalizations. The reporters are working at home, which was never experienced before the Covid-19 pandemic, and they have difficulties in covering their reporting beats because they have little or no access to contact points. The reporters rely heavily on indirect coverage and online briefings via phones or SNS because they have difficulties in meeting news sources in person. As a result, the diversity of news contents and the media's on-site monitoring functions has been severely weakened. In addition, the reporters have no chances to both exchange ideas with fellow reporters and to transfer the know-how of collecting news items to their juniors. This paper has also discussed the disruption of practices that the ongoing Covid-19 has brought to the media ecosystem from a variety of perspectives.

Hsa_Circ_0001947/MiR-661/DOK7 Axis Restrains Non-Small Cell Lung Cancer Development

  • Bao, Yuyan;Yu, Yanjie;Hong, Bing;Lin, Zhenjian;Qi, Guoli;Zhou, Jie;Liu, Kaiping;Zhang, Xiaomin
    • Journal of Microbiology and Biotechnology
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    • v.31 no.11
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    • pp.1508-1518
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    • 2021
  • Hsa_circ_0001947 is associated with multiple cancers, but its function in non-small cell lung cancer (NSCLC) is ambiguous and needs further research. The targeting relationship among circ_0001947, miR-661, and downstream of tyrosine kinase 7 (DOK7) was predicted by database and further verified by dual-luciferase reporter assay, while their expressions in cancer tissues and cells were detected by quantitative real-time polymerase chain reaction (qRT-PCR). After transfection, cell biological behaviors and expressions of miRNAs, miR-661 and DOK7 were determined by cell function experiments and qRT-PCR, respectively. Circ_0001947 was low-expressed in NSCLC tissues and cells. Circ_0001947 knockdown intensified cell viability and proliferation, induced cell cycle arrest at S phase, suppressed apoptosis and evidently enhanced miR-510, miR-587, miR-661 and miR-942 levels, while circ_0001947 overexpression did the opposite. MiR-661 was a target gene of circ_0001947 that participated in the regulation of circ_0001947 on cell biological behaviors. Furthermore, DOK7, the target gene of miR-661, partly participated in the regulation of miR-661 on cell viability. Hsa_circ_0001947 acts as a sponge of miR-661 to repress NSCLC development by elevating the expression of DOK7.