• 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.

• 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.

• Tuberculosis and Respiratory Diseases
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• 제60권6호
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• pp.663-672
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• 2006

연구배경: 미세먼지는 여러 가지 유기물과 무기물의 복합체로 그 구성 성분이 시간과 장소에 따라 다르고 모양과 크기도 일정하지 않으며, 특히 지름 $10{\mu}m$이하의 미세먼지 (particulate matter 10; $PM_{10}$)는 흡입이 가능한 입자의 크기여서 하부기관지 및 폐의 가스-교환부분까지 침착하여 호흡기계에 손상을 일으킬 수 있다. 본 연구에서는 황사에 포함된 $PM_{10}$과 비황사 시기에 포집된 $PM_{10}$이 폐상피세포주에 작용하여 전염증성 사이토카인(proinflammatory cytokine) 및 cytokine messenger RNA(mRNA)의 발현에 어떤 영향이 있는지를 관찰하여 기관지 천식과 만성 폐쇄성 폐질환등 호흡기 질환의 증상 악화기전에 미치는 역할을 규명하고자 하였다. 연구방법: 공기 포집기(HV 500F, sibata model)를 이용하여 황사와 비황사 기간에 하루 6시간씩 실외의 장소에서 대기분진을 membrane filter에 포집한 다음, $PM_{10}$입자를 추출하고 폐암 상피세포주인 A549 cells(한국세포은행주)에 $PM_{10}$을 농도에 맞게($10{\mu}g/ml$, $100{\mu}g/ml$, $500{\mu}g/ml$) 노출시켰다. 각각의 노출된 세포로부터 interleukin(IL)-$1{\alpha}$, $IL-1{\beta}$, IL-8, granulocyte macrophage colony stimulating factor(GM-CSF)의 mRNA를 역전사중합효소연쇄반응(reverse transcriptase polymerase chain reaction; RT-PCR) 방법으로 측정하였다. 결 과: 황사 및 비황사 기간 중 포집된 $PM_{10}$을 가했을 시 가하지 않은 대조군에 비하여 $IL-1{\alpha}$, $IL-1{\beta}$, IL-8, granulocyte macrophage colony stimulating factor (GM-CSF)의 m'RNA와 cytokine의 발현이 유의하게 높았으며, 황사 기간의 고농도의 $PM_{10}$에 노출된 세포의 $IL-1{\alpha}$ mRNA는 비황사 기간의 $PM_{10}$에 노출된 세포의 mRNA보다 증가되어 있었다. 결 론: $PM_{10}$은 A549 cells에서 전염증성 사이토카인의 발현을 증가시키고 비황사 기간보다 황사 기간 중 대기 중에서 채취한 $PM_{10}$에 노출된 A549 cells에서 일부의 전염증성 사이토카인의 mRNA발현을 더욱 증가시키는 것을 알 수 있었다. 따라서 황사 기간의 $PM_{10}$에 의한 일부의 전염증성 사이토카인의 발현 증가가 만성 호흡기 질환의 증상 악화기전에 연관되어 있을 가능성을 시사하였다.