• Animal Bioscience
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• 제34권5호
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• pp.801-810
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• 2021

Objective: microRNAs (miRNAs) can play a role in a variety of physiological and pathological processes, and their role is achieved by regulating the expression of target genes. Our previous high-throughput sequencing found that ssc-miR-185 plays an important regulatory role in piglet diarrhea, but its specific target genes and functions in intestinal porcine epithelial cell (IPEC-J2) are still unclear. We intended to verify the target relationship between porcine miR-185 and cell division cycle 42 (CDC42) gene in IPEC-J2 and to explore the effect of miR-185 on the proliferation of IPEC-J2 cells. Methods: The TargetScan, miRDB, and miRanda software were used to predict the target genes of porcine miR-185, and CDC42 was selected as a candidate target gene. The CDC42-3' UTR-wild type (WT) and CDC42-3'UTR-mutant type (MUT) segments were successfully cloned into pmirGLO luciferase vector, and the luciferase activity was detected after co-transfection with miR-185 mimics and pmirGLO-CDC42-3'UTR. The expression level of CDC42 was analyzed using quantitative polymerase chain reaction and Western blot. The proliferation of IPEC-J2 was detected using cell counting kit-8 (CCK-8), methylthiazolyldiphenyl-tetrazolium bromide (MTT), and 5-ethynyl-2'-deoxyuridine (EdU) assays. Results: Double enzyme digestion and sequencing confirmed that CDC42-3'UTR-WT and CDC42-3'UTR-MUT were successfully cloned into pmirGLO luciferase reporter vector, and the luciferase activity was significantly reduced after co-transfection with miR-185 mimics and CDC42-3'UTR-WT. Further we found that the mRNA and protein expression level of CDC42 were down-regulated after transfection with miR-185 mimics, while the opposite trend was observed after transfection with miR-185 inhibitor (p<0.01). In addition, the CCK-8, MTT, and EdU results demonstrated that miR-185 promotes IPEC-J2 cells proliferation by targeting CDC42. Conclusion: These findings indicate that porcine miR-185 can directly target CDC42 and promote the proliferation of IPEC-J2 cells. However, the detailed regulatory mechanism of miR-185/CDC42 axis in piglets' resistance to diarrhea is yet to be elucidated in further investigation.

• Molecules and Cells
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• 제40권3호
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• pp.195-201
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• 2017

In this study, qRT-PCR was employed to identify that miR-186 expression level in NSCLC tissues are highly associated with lymph node metastasis. In addition, through the application of western blotting, luciferase assay and qRT-PCR, it was found that miR-186 targeted 3'UTR of cdc42 mRNA and down-regulated cdc42 protein level in a post-transcriptional manner. Transwell assay indicated that cdc42 partially reversed the effect of miR-186 mimics. Besides, miR-186 was proved to regulate EMT by influencing biomarkers of this process and cell adhesion ability. Thus, miR-186 is a potential target for NSCLC therapy. miR-186 is proposed to be one of tumor-suppressors and may serve as a therapeutic target in NSCLC treatment.

• Biomolecules & Therapeutics
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• 제22권6호
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• pp.540-546
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• 2014

The high mortality rates associated with cancer reflect the metastatic spread of tumor cells from the site of their origin. Metastasis, in fact, is the cause of 90% of cancer deaths. Therefore, considerable effort is being made to inhibit metastasis. In the present study, we screened ketotifen for anti-migratory and anti-invasive activities against MDA-MB-231 breast cancer and HT-1080 fibrosarcoma cancer cells. Cancer cell migration and invasion were measured using multi-well chambers. Additionally, western blots were used to examine the effects of ketotifen on the expressions of CDC42, Rho, Rac, and matrix metalloproteinase 9 (MMP-9). The results showed that ketotifen dose-dependently suppressed the migration and invasion of MDA-MB-231 and HT-1080 cells. Ketotifen also suppressed the expressions of CDC42, Rac, and Rho, which, significantly, are involved in MDA-MB-231 and HT-1080 cancer cell migration. Moreover, ketotifen suppressed the expression and activity of MMP-9, which is involved in degradation of the extracellular matrix leading to invasion. The overall data suggested that ketotifen suppresses the migration and invasion of MDA-MB-231 and HT-1080 cancer cells via inhibition of CDC42, Rac, Rho, and MMP-9 expression.

• 대한임상검사과학회지
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• 제43권3호
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• pp.124-132
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• 2011

The aim of this study was to evaluate the effects of the photosensitizer photogem with light-emitting diode (LED) on vancomycin-resistant enterococci (VRE). Two VRE strains isolated from the feces of patients. that was identificated Enterococcus faecium (vanA) and Enterococcus gallinarum (vanC1) using traditional biochemical tests and confirmed VRE genotyping from using polymerase chain reaction. In addition, three strains were used Enterococcus. faecalis CDC-286 (vanA), E. faecalis CDC-583 (vanB) and E. gallinarum CDC-42 (vanC1). To examine the antimicrobial effect of photogem mediated photodynamic therapy (PDT) against, CFU quantification and Disk diffusion antimicrobial susceptibility test were evaluated. The effects of Photodynamic therapy was not associated with genotype. Photogem mediated PDT perfectly inhibited the colony formation of E. faecalis CDC-286. The number of viable bacteria decreased greatly after PDT application with photogem $50{\mu}g/mL$ and energy density of $15J/cm^2$. The diameter of inhibition zone was increased to after PDT more than before PDT. The case of vancomycin disc on E. faecalis CDC-583 and E. galinanum-Patient were changed from resistant to intermediate resistant, from intermediate resistant to susceptable. These results demonstrate that lethal photosensitization of VRE can be achieved using photogem plus 630 nm LED irradiation.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.222.1-222.1
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• 2003

The signal transduction pathway leading to the activation of the p70s6k plays an important role in the progression of cells from G0/G1 to S phase of the cell cycle but remains incompletely characterized. We investigated the role of the Rho family G protein Rac1 in H2O2-mediated p70s6k activation. Transient expression of a dominant negative mutants of the small GTP-binding proteins Rac1 (Rac1N17) and Cdc42(Cdc42N17) showed reduced levels of slower migration on Western blots of one-dimensional SDS-PAGE in p70s6k and ERK1/2 by PDFG stimulation. (omitted)

• 한국미생물·생명공학회지
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• 제42권1호
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• pp.73-81
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• 2014

본 연구에서는 명아주과 수송나물속 솔장다리(Salsola collina Pall.) 추출물의 항산화 및 항암 활성을 분석하였다. 먼저 솔장다리의 에탄올 추출물의 DPPH radical scavenging activity를 분석한 결과, $IC_{50}$가 $4.82{\mu}g/ml$로 나타나 강한 항산화능을 보유하였음을 확인하였다. 또한 대장암 세포주(HT29), 폐암 세포주(A549), 간암 세포주(HepG2)를 사용하여 솔장다리 추출물의 암세포 사멸효과를 분석한 결과, $IC_{50}$가 각각 43.8, 64.1, $92.5{\mu}g/ml$로 강력한 세포사멸효과를 나타냈으며 특히 HT29에 대한 강한 사멸효과를 보였다. 솔장다리 추출물의 항암 활성 기전 분석을 위해 세포주기를 분석한 결과, 대장암세포인 HT29의 G2/M arrest를 유도하였으며 최고 농도인 $60{\mu}g/ml$까지 S기 세포수가 증가하였다. 세포주기관련 단백질의 발현 분석 결과, 솔장다리 추출물을 처리한 경우, G2기에서 M기로의 전이에 필수적인 단백질인 Cdc25C와 cyclin A의 발현이 감소되었고, 반면 Cdc25C와 Cdc2의 불활성화 형태인 p-Cdc25C, p-Cdc2는 증가하였다. 또한 p21과 Wee1의 발현도 증가되었다. 하지만 p53의 발현량은 변화가 없었다. 이러한 결과는 솔장다리 추출물을 처리한 경우, p53 비의존적으로 p21의 발현이 증가되어 cyclin A/Cdc2 complex의 활성이 조절되고, 이어서 G2/M phase의 check point에 작용하는 Wee1의 발현증가 및 Cdc25C, Cdc2의 인산화에 의한 불활성화를 통하여 G2/M arrest가 유도되는 것을 시사한다. 또한 솔장다리 추출물 처리에 의해 S기 진행을 조절하는 Cdk2의 발현량도 감소하여, cyclin A/Cdk2 complex가 감소되어 S기의 세포수가 증가한 것으로 보인다. 따라서 본 연구 결과를 통해 솔장다리 추출물이 높은 항산화 활성을 지니며 암세포의 세포주기를 조절하여 높은 항암 활성을 보유함을 확인하였다.

• Molecules and Cells
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• 제26권5호
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• pp.462-467
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• 2008

TPA is known to cooperate with an activated Ras oncogene in the transformation of rodent fibroblasts, but the biochemical mechanisms responsible for this effect have not been established. In the present study we used c-fos promoter-luciferase constructs as reporters, in transient transfection assays, in NIH3T3 cells to assess the mechanism of this cooperation. We found a marked synergistic interaction between TPA and a transfected v-Ha-ras oncogene in the activation of c-fos promoter and SRE. SRE has binding sites for TCF and SRF. A dominant-negative Ras (ras-N17) inhibited the TPA-Ras synergy by blocking the PKC-MAPK-TCF pathway. Dominant-negative RhoA and Rac1 (but not Cdc42Hs) inhibited the TPA-Ras synergy by blocking the Ras-Rho-SRF signaling pathway. Constitutively active $PKC{\alpha}$ and $PKC{\varepsilon}$ showed synergy with v-Ras. These results suggest that the activation of two distinct pathways such as Ras-Raf-ERK-TCF pathway and Rho-SRF pathway are responsible for the induction of c-fos by TPA and Ras in mitogenic signaling pathways.

• 미생물학회지
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• 제33권4호
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• pp.232-236
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• 1997

Septin 유전자는 Saccharomyces cerevisiae에서 filament를 암호화하고 있으며 세포질분열이나 bud의 형성에 중요한 역할을 하는 것으로 알려져있다. S. cerevisiae에서 septin유전자는 4가지가 발견되었으며 초파리나 쥐의 세포에서도 발견되고 있다. 본 연구에서는 PCR 방법을 이용하여 Schizosaccharomyces pombe에서 septin 유전자를 찾아내었다. S. pombe의 septin 유전자는 1143 bp의 open reading frame을 갖고 있으며 380개의 아미노산으로된 42 kd의 분자량을 가진 단백질을 암호화하였다. S. cerevisiae의 septin 유전자의 하나인 $CDC_{12}$ 유전자와의 유사성을 비교한 결과 51.8%의 유사성이 있음이 밝혀졌다.

• 보건교육건강증진학회지
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• 제22권4호
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• pp.57-72
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• 2005

Objectives: To investigate the status of health education in elementary schools. Methods: 620 school nurses were surveyed by questionnaires from September to December in 2003. Results: 1) Among the school nurses under inquiry of planning of health education, 3.3% and 9.3% of them did not prepare for teaching plan. 2) The average time for health education by a school nurse was 96.8 hours a year, and handouts for health education were distributed 10.6 times. Among the contents of health education, sex education took the largest portion of health education with 24.6 hours a year. 3) With regard to the contents of health education covered by school nurses, sex education ranked first with 90.2%, next came drug abuse with 78.4%, dental health, CDC, disease control, healthy life and smoking, body structure and function and growth and development safety, alcohol, nutrition and environmental health followed them. 4) The main contents of education were CDC, dental health sex, healthy life and disease control for 1st, 2nd and 3rd grade students, sex, CDC, disease control and safety for 4th grade students, and sex, CDC, drug abuse and smoking for 5th and 6th grade students. 5) 72.6% of school nurses used class room for health education, 20.0% and 7.4% of them used grade and others such as broadcast, respectively. 6) 42.1% of school nurses used blackboard, 37.0% and 18.6% of them used visual media and handout as a teaching aids for health education. 7) 31.6% of school nurses replied that education time was insufficient 9.5% and 15.9% of them replied the contents of health education were inadequate and methods of health education were inappropriate, respectively. Conclusions: For the successful school health education, it would be in need of sufficient time for health education by opening health education course and of modify the various working conditions of school nurses, and those of effective educational materials and media for health education.

• Nutrition Research and Practice
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• 제14권2호
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• pp.127-133
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• 2020

BACKGROUND/OBJECTIVES: Non-small cell lung cancer is mostly recognized among other types of lung cancer with a poor prognosis by cause of chemotherapeutic resistance and increased metastasis. Luteolin has been found to decrease cell metastasis. However, its underlying mechanisms remain unresolved. The objective of this study was to examine the effect (and its mechanism) of luteolin on the migration and invasion of human non-small cell lung cancer A549 cells. MATERIALS/METHODS: Cell viability was investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Wound healing and transwell assays were evaluated to assess migration and invasion, respectively. Western blot analysis and immunofluorescence were further performed to investigate the role of luteolin and its mechanisms of action. RESULTS: Administration with up to 40 μM luteolin showed no cytotoxic activity on lung cancer A549 cells or non-cancer MRC-5 cells. Additionally, luteolin at 20-40 μM significantly suppressed A549 cells' migration, invasion, and the formation of filopodia in a concentration-dependent manner at 24 h. This is similar with western blot analysis, which revealed diminished the phosphorylated focal adhesion kinase (pFAK), phosphorylated non-receptor tyrosine kinase (pSrc), Ras-related C3 botulinum toxin substrate 1 (Rac1), cell division control protein 42 (Cdc42), and Ras homolog gene family member A (RhoA) expression levels. CONCLUSIONS: Overall, our data indicate that luteolin plays a role in controlling lung cancer cells' migration and invasion via Src/FAK and its downstream Rac1, Cdc42, and RhoA pathways. Luteolin might be considered a promising candidate for suppressing invasion and metastasis of lung cancer cells.