• Asian Pacific Journal of Cancer Prevention
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• 제15권21호
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• pp.9131-9136
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• 2014

ELDF15, homologous with AT2 receptor-interaction protein 1 (ATIP1), may play an important role in cell differentiation, proliferation, and carcinogenesis. We aimed to understand the biological function of ELDF15 via construction and transfection of a recombinant expression vector containing antisense ELDF15. Recombinant expression vectors were successfully constructed and transfected into K562 cells. A stable transfectant, known as pXJ41-asELDF15, stably produced antisense ELDF15. Compared with K562 and K562-zeo cells, K562-pXJ41-asELDF15 cells showed inhibition of cell proliferation. RT-PCR analysis showed that the expression and protein level of ELDF15 decreased significantly in K562 cells transfected with pXJ41-asELDF15. Expression of hemoglobin increased in K562 cells transfected with pXJ41-asELDF15 by benzidine staining. increases NBT reduction activity in K562 cells transfected with pXJ41-asELDF15.Colony forming efficiency in two-layer soft agar was clearly inhibited as assessed by electron microscopy. These results suggest that ELDF15 plays a potential role in cell differentiation, proliferation and carcinogenesis.

• Asian Pacific Journal of Cancer Prevention
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• 제13권12호
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• pp.5975-5979
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• 2012

Background: To investigate the influence of telomerase activity, apoptosis, radiosensitivity of cervical cancer after siRNA-mediated knockdown of telomerase RNA and evaluate in vivo growth with gene interference. Methods: We studied siRNA-targeting-telomerase RNA transfection into the Hela cell line. Expression of hTERC mRNA was detected by RT-PCR and telomerase activity was measured by the TRAP assay. Growth inhibition was determined by MTT assay and radiosensitivity of the cervical cancer cells was examined by colony formation assay. In addtion, effects of hTERC inhibition in vivo were studied by injection of siRNA-transfected Hela cells into nude mice. Results: The hTERC siRNA effectively downregulated the expression of hTERC mRNA and also reduced the telomerase activity to 30% of the untreated control vlaue. The viability of hTERC siRNA transfected Hela cells was reduced by 44.7% after transfection. After radiation treatment, the radiosensitivity of Hela cells with hTERC knockdown was increased. In vivo, the tumors developing from the hTERC siRNA-transfected cells were of reduced size, indicating that the hTERT siRNA also depressed the tumorigenic potential of the Hela cells. Conclusions: Our results supported the concept of siRNA-mediated inhibition of telomerase mRNA which could inhibit the expression of hTERC and telomerase activity. Furthermore, radiosensitivity was upregulated after knockdown the hTERC in vivo and in vitro.

• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.836-843
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• 2004

Nontypeable H. influenzae (NTHi), a Gram-negative obligate human pathogen, causes pneumonia, chronic bronchitis, and otitis media, and the respiratory epithelium is the first line of defense that copes with the pathogen. In an effort to identify transcriptional responses of human respiratory epithelial cells to infection with NTHi, we examined its differential gene expression using high density cDNA microarrays. BEAS-2B human bronchial epithelial cells were exposed to NTHi for 3 hand 24 h, and the alteration of mRNA expression was analyzed using microarrays consisting of 8,170 human cDNA clones. The results indicated that approximately 2.6% of the genes present on the microarrays increased in expression over 2-fold and 3.8% of the genes decreased during the 24-h infection period. Upregulated genes included cytokines (granulocyte-macrophage colony stimulating factor 2, granulocyte chemotactic protein 2, IL-6, IL-10, IL-8), transcription factors (Kruppel-like factor 7, CCAAT/enhancer binding protein $\beta$, E2F-1, NF-$\kappa$B, cell surface molecules (CD74, ICAM-1, ICAM-2, HLA class I), as well as those involved in signal transduction and cellular transport. Selected genes were further confirmed by reverse-transcription-PCR. These data expand our knowledge of host cellular responses during NTHi infection and should provide a molecular basis for the study of host-NTHi interaction.

• Journal of Microbiology and Biotechnology
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• 제12권5호
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• pp.716-721
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• 2002

The ldhL gene encoding the $_L$-(+) lactate dehydrogenase was cloned from Lactobacillus reuteri ATCC 55739 chromosomal DNA and characterized. An internal 750-bp fiagment of ldhL gene was amplified by PCR using primers based on the conserved region of lactobacilli ldhL genes. A genomic library off. reuteri ATCC 55739 was constructed using pBR322, and colony hybridization experiments were performed using the 750-bp fragment as aprobe. One clone harboring a 4.0-kb PstI fragment was identified, and nucleotide sequencing confirmed it as an open reading frame of 972 bp in size in the middle. In addition to IdhL gene, an ORF homologous to Streptococcus pneumoniae TIGR4 hydrolase gene and 3' part of phosphomevalonate kinase gene (mvaK2) were also found on the 4 kb fragment. $_L$-LDH of L. reuteri ATCC 55739 showed the highest degree of homology with the $_L$-LDH of Pediococcus acidilactici (62.4%), fullowed by the $_L$-LDH of Lactobacillus pentosus (58.7%). The size of IdhL transcript determined by Northern blot was 1 kb, indicating the monocistronic nature of IdhL.

• Journal of Microbiology
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• 제38권4호
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• pp.230-238
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• 2000

Class III chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class III chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5'flanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at -716 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for past-transnational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P chrysogenum and genus Aspergillus.

• 한국환경과학회지
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• 제30권5호
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• pp.369-378
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• 2021

In this study, we investigated the effects of indole on biofilm formation inhibition in Pantoea agglomerans (P. agglomerans). In the biofilm growth assay, indole inhibited biofilm formation across all the growth time. Depending on biofilm growth stage, indole exhibited biofilm inhibition and anti-bacterial effects on planktonic cells. Through the analysis of the proportion rate between biofilm and Colony Forming Units (CFU) and inhibition rate of indole, we confirmed that depending on the biofilm stage of P. agglomerans, indole treatment timing was more important than the treatment duration. By comparing gene expression rates through rt-qPCR P.agglomerans affected by indole was found to significantly change quorum sensing (pagI/R) and indole transportation (bssS) gene expressions. Throughout all, indole exhibited both antimicrobial and anti-biofilm effects on P. agglomerans. In addition, we confirmed the anti-biofilm effects of indole on mature biofilm. In conclusion, indole as a signal molecule, can exhibit anti-biofilm effects through bacterial quorum sensing inhibition and indole affects. Therefore, indole can regulate biofilm bacteria especially gram-negative opportunistic pathogens.

• Journal of Microbiology and Biotechnology
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• 제31권6호
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• pp.784-793
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• 2021

Previous studies have uncovered the role of circ_0000144 in various tumors. Here, we investigated the function and mechanism of circ_0000144 in gastric cancer (GC) progression. The expression of circ_0000144 in GC tissues and cells was detected through quantitative real-time polymerase chain reaction (qRT-PCR) method. Gain- and loss-of-function experiments including colony formation, wound healing and transwell assays were performed to examine the role of circ_0000144 in GC cells. Furthermore, western blot was conducted to determine the expressions of epithelial mesenchymal transition (EMT)-related proteins. The interaction between circ_0000144 and miR-217 was analyzed by bioinformatic analysis and luciferase reporter assays. The circ_0000144 expression was obviously upregulated in GC tissues and cells. Silencing of circ_0000144 inhibited cell proliferation, migration and invasion of GC cells, but ectopic expression of circ_0000144 showed the opposite results. Moreover, circ_0000144 sponged miR-217, and rescue assays revealed that silencing miR-217 expression reversed the inhibitory effect of circ_0000144 knockdown on the progress of GC. Our findings reveal that circ_0000144 inhibition suppresses GC cell proliferation, migration and invasion via absorbing miR-217, providing a new biomarker and potential therapeutic target for treatment of GC.

• 한국균학회지
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• 제51권4호
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• pp.491-504
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• 2023

To explore fungal diversity in orchard soil where fire-blighted apple trees are buried, we collected soil samples from apple orchards in Chungju, Korea. Fungal isolates were obtained from DG18 agar and identified at the species level based on morphological features and phylogenetic analyses. The colony characteristics and microstructures were examined using a light microscope and a scanning electron microscope after culturing on potato dextrose agar (PDA), malt extract agar (MEA), Czapek yeast agar (CYA), and oatmeal agar (OA) The PCR-amplified products of the ITS1-5.8S-ITS2 region and 28S large subunit of the nuclear ribosomal RNA gene, as well as partial sequences of the β-tubulin, calmodulin, and translation elongation factor 1-α genes were sequenced and analyzed phylogenetically. Seven previously unknown fungal species were explored in Korea. All samples, including Aspergillus aureolatus, Botryotrichum atrogriseum, Dactylonectria novozelandica, Fusarium denticulatum, Paecilomyces tabacinus, Sarcopodium tibetense and Talaromyces stollii, had ascomycetes. Herein, we report their descriptions and features.

• The Korean Journal of Physiology and Pharmacology
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• 제28권4호
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• pp.379-387
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• 2024

Breast cancer (BC) is most commonly diagnosed worldwide. Liquiritigenin is a flavonoid found in various species of the Glycyrrhiza genus, showing anti-tumor activity. This article was to explore the influences of liquiritigenin on the biological behaviors of BC cells and its underlying mechanism. BC cells were treated with liquiritigenin alone or transfected with oe-HSP90 before liquiritigenin treatment. RT-qPCR and Western blotting were employed to examine the levels of HSP90, Snail, E-cadherin, HSC70, and LAMP-2A. Cell viability, proliferation, migration, and invasion were evaluated by performing MTT, colony formation, scratch, and Transwell assays, respectively. Liquiritigenin treatment reduced HSP90 and Snail levels and enhanced E-cadherin expression as well as inhibiting the proliferation, migration, and invasion of BC cells. Moreover, liquiritigenin treatment decreased the expression of HSC70 and LAMP-2A, proteins related to chaperone-mediated autophagy (CMA). HSP90 overexpression promoted the CMA, invasion, and migration of BC cells under liquiritigenin treatment. Liquiritigenin inhibits HSP90-mediated CMA, thereby suppressing BC cell growth.

• 한국수정란이식학회지
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• 제21권1호
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• pp.13-20
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• 2006

본 연구의 목적은 요를 통해 hFSH를 발현하는 형질 전환 소의 생산이다. 요의 분비와 관련 있는 유전자로서 mUII promoter를 사용하여 hFSH유전자를 구성했다. 태아섬유아세포(KbFF)는 임신 45일령의 태아(male)에서 채취하였다. hFSH gene은 pcDNA3(neo) vector와 같이 KbFF 세포에 electroporation 방법으로 transfection하였다. 유전자를 transfection한 세포는 G-418로 2주 동안 배양하였고, 선발된 colony는 PCR로 확인하였다. 핵이 제거된 난자는 hFSH가 transfection된 세포와 transfection 되지 않은(control) 세포를 이용하여 핵이식하였다. 48시 간 후 hFSH가 transfection된 세포는 68.7%의 수정란이 난할되었으며, 8일 후 15.7%의 수정란이 배반포로 발달하였다. 그러나 대조구에서는 67.6%가 난할되었으며, 24.5%가 배반포로 각각 발달하였다. 이들 배반포에서 apoptosis 분석 결과 hFSH 유전자가 transfection된 또는 transfection되지 않은 대조구에서 유의적인 차이는 보이지 않았다. 배반포는 53두의 수란우에 이식하여 두 마리의 산자가 생산되었으나(1.9%) hFSH가 transfection되지 않은 것으로 나타났다. 이 결과는 선발된 hFSH colony에서 transfection되지 않은 세포가 혼합되어 있었다는 것을 나타내 주고 있으며, colony의 선발과 검증에 더 많은 연구의 필요성이 있음을 나타내준다.