• Title/Summary/Keyword: PLK4

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Inhibition of Polo-like Kinase 1 Prevents the Male Pronuclear Formation Via Alpha-tubulin Recruiting in In vivo-fertilized Murine Embryos

  • Moon, Jeonghyeon;Roh, Sangho
    • Journal of Embryo Transfer
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    • v.33 no.4
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    • pp.229-235
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    • 2018
  • Polo-like kinase 1 (Plk1) has been known to be a critical element in cell division including centrosome maturation, cytokinesis and spindle formation in somatic, cancer, and mammalian embryonic cells. In particular, Plk1 is highly expressed in cancer cells. Plk1 inhibitors, such as BI2536, have been widely used to prevent cell division as an anticancer drug. In this study, the fertilized murine oocytes were treated with BI2536 for 30 min after recovery from the oviduct to investigate the effect of down-regulation of Plk1 in the in vivo-fertilized murine embryos. Then, the localization and expression of Plk1 was observed by immunofluorescence staining. The sperm which had entered into the oocyte cytoplasm did not form male pronuclei in BI2536-treated oocytes. The BI2536-treated oocytes showed significantly lower expression of Plk1 than non-treated control group. In addition, alpha-tubulin and Plk1 gathered around sperm head in non-treated oocytes, while BI2536-treated oocytes did not show this phenomenon. The present study demonstrates that the Plk1 inhibitor, BI2536, hinders fertilization by inhibiting the formation of murine male pronucleus.

Relation between Cyclooxygenase-2 and Polo-like Kinase-1 in Non-Small Cell Lung Cancer (비소세포 폐암에서 Cyclooxygenase-2와 Polo-like Kinase-1의 상관관계)

  • Lee, Kyu-Hwa;Yang, Seok-Chul
    • Tuberculosis and Respiratory Diseases
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    • v.67 no.4
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    • pp.303-310
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    • 2009
  • Background: Elevated expression of cyclooxygenase-2 (COX-2) and Polo-like kinase-1 (PLK-1) is observed in a wide variety of cancers. Augmented expression of COX-2 and enhanced production of prostaglandin $E_2(PGE_2)$ are associated with increased tumor cell survival and malignancy; COX-2 has been implicated in the control of human non-small cell lung carcinoma (NSCLC) cell growth. PLK-1 siRNA induced the cell death of lung cancer cells and the systemic administration of PLK-1 siRNA/atelocollagen complex inhibited the growth of lung cancer in a liver metastatic murine model. COX-2 and PLK-1 are involved in proliferation and in cell cycle regulation, and there is a significant correlation between their interaction in prostate carcinoma. Methods: In this study, we investigated the pattern of COX-2 and PLK-1 expression in NSCLC, after treatment with IL-1$\beta$, COX-2 inhibitor and PLK-1 siRNA. Results: Expression of PLK-1 was decreased in A549 COX-2 sense cells, and was increased in A549 COX-2 anti-sense cells. Knock out of PLK-1 expression by PLK-1 siRNA augmented COX-2 expression in A549 and NCl-H157 cells. When A549 and NCI-H157 cells were treated with COX-2 inhibitor on a dose-dependent basis, PLK-1 and COX-2 were reduced. However, when the expression of COX-2 was induced by IL-1$\beta$, the production of PLK-1 decreased. Conclusion: These results demonstrate that COX-2 and PLK-1 are regulated and inhibited by each other in NSCLC, and suggest that these proteins have a reverse relationship in NSCLC.

Mechanisms Underlying Plk1 Polo-Box Domain-Mediated Biological Processes and Their Physiological Significance

  • Lee, Kyung S.;Park, Jung-Eun;Kang, Young Hwi;Kim, Tae-Sung;Bang, Jeong K.
    • Molecules and Cells
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    • v.37 no.4
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    • pp.286-294
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    • 2014
  • Mammalian polo-like kinase 1 (Plk1) has been studied intensively as a key regulator of various cell cycle events that are critical for proper M-phase progression. The polobox domain (PBD) present in Plk1's C-terminal noncatalytic region has been shown to play a central role in targeting the N-terminal kinase domain of Plk1 to specific subcellular locations. Subsequent studies reveal that PBD binds to a phosphorylated motif generated by one of the two mechanisms - self-priming by Plk1 itself or non-self-priming by a Pro-directed kinase, such as Cdc2. Here, we comparatively review the differences in the biochemical steps of these mechanisms and discuss their physiological significance. Considering the diverse functions of Plk1 during the cell cycle, a better understanding of how the catalytic activity of Plk1 functions in concert with its cisacting PBD and how this coordinated process is intricately regulated to promote Plk1 functions will be important for providing new insights into different mechanisms underlying various Plk1-mediated biological events that occur at the multiple stages of the cell cycle.

PLK2 Single Nucleotide Variant in Gastric Cancer Patients Affects miR-23b-5p Binding

  • Dominkus, Pia Puzar;Mesic, Aner;Hudler, Petra
    • Journal of Gastric Cancer
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    • v.22 no.4
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    • pp.348-368
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    • 2022
  • Purpose: Chromosomal instability is a hallmark of gastric cancer (GC). It can be driven by single nucleotide variants (SNVs) in cell cycle genes. We investigated the associations between SNVs in candidate genes, PLK2, PLK3, and ATM, and GC risk and clinicopathological features. Materials and Methods: The genotyping study included 542 patients with GC and healthy controls. Generalized linear models were used for the risk and clinicopathological association analyses. Survival analysis was performed using the Kaplan-Meier method. The binding of candidate miRs was analyzed using a luciferase reporter assay. Results: The PLK2 Crs15009-Crs963615 haplotype was under-represented in the GC group compared to that in the control group (Pcorr=0.050). Male patients with the PLK2 rs963615 CT genotype had a lower risk of GC, whereas female patients had a higher risk (P=0.023; P=0.026). The PLK2 rs963615 CT genotype was associated with the absence of vascular invasion (P=0.012). The PLK3 rs12404160 AA genotype was associated with a higher risk of GC in the male population (P=0.015). The ATM Trs228589-Ars189037-Grs4585 haplotype was associated with a higher risk of GC (P<0.001). The ATM rs228589, rs189037, and rs4585 genotypes TA+AA, AG+GG, and TG+GG were associated with the absence of perineural invasion (P=0.034). In vitro analysis showed that the cancer-associated miR-23b-5p mimic specifically bound to the PLK2 rs15009 G allele (P=0.0097). Moreover, low miR-23b expression predicted longer 10-year survival (P=0.0066) in patients with GC. Conclusions: PLK2, PLK3, and ATM SNVs could potentially be helpful for the prediction of GC risk and clinicopathological features. PLK2 rs15009 affects the binding of miR-23b-5p. MiR-23b-5p expression status could serve as a prognostic marker for survival in patients with GC.

Understanding centrosome amplification in cancer: A pathway toward precision-targeted cancer drug development (암의 중심체 증폭 이해를 통한 표적 항암제 개발)

  • Taekyung Kim
    • Journal of Life Science
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    • v.33 no.11
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    • pp.950-955
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    • 2023
  • Cell division is an essential process for the survival and development of living organisms. It is critical that duplicated chromosomes are properly segregated into daughter cells during mitosis. The centrosome is the core organelle that forms the microtubule-organizing center (MTOC), which generates the microtubules that make up the mitotic spindle during cell division. The centrosome is also involved in cell signaling and motility. In normal cells, there is one centrosome in G1 that replicates into two in the S phase and matures through G2. During the M phase, duplicated centrosomes move to both ends of the cell, and spindle microtubules that are generated from MTOC move the chromosome to both ends. The cells then split into two to complete the cell division. However, a phenomenon called centrosome amplification (CA), in which the number of centrosomes is higher than normal, is common in cancer cells and can lead to chromosome instability (CIN). This paper discusses the process of centrosome replication and the role of PLK4 in this process. The possible consequences of centrosome amplification and how the PLK4 inhibitor may be able to treat certain types of cancer cells, such as breast cancer and neuroblastoma, will also be discussed.

Ectopic Expression of Cenexin1 S796A Mutant in $ODF2^{+/-}$ Knockout Background Causes a Sperm Tail Development Defect

  • Lee, Kyung Ho
    • Development and Reproduction
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    • v.16 no.4
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    • pp.363-370
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    • 2012
  • The outer dense fiber 2 (ODF2) protein is an important component of sperm tail outer dense fiber and localizes at the centrosome. It has been reported that the RO072 ES cell derived homozygote knock out of ODF2 results in an embryonic lethal phenotype, and XL169 ES cell derived heterozygote knock out causes severe defects in sperm tail development. The ODF2s splicing variant, Cenexin1, possesses a C-terminal extension, and the phosphorylation of serine 796 residue in an extended C-terminal is responsible for Plk1 binding. Cenexin1 assembles ninein and causes ciliogenesis in early stages of the cell cycle in a Plk1-independent manner. Alternatively, in the late stages of the cell cycle, G2/M phase, Cenexin1 binds to Plk1 and results in proper mitotic progression. In this study, to identify the in vivo function of Plk1 binding to phosphorylated Cenexin1 S796 residue, and to understand the in vivo functional differences between ODF2 and Cenexin1, we generated ODF2/Cenexin1 S796A/Cenexin1 WT expressing transgenic mice in a RO072 ES cell derived $ODF2^{+/-}$ knock out background. We observed a severe defect of sperm tail development by ectopic expression of Cenexin1 S796A mutant and no phenotypic differences between the ectopic expression of ODF2/Cenexin1 WT in $ODF2^{+/-}$ background and in normal wild type mice.

Inhibitory effects of Sanghwang mushrooms, Phellinus linteus strains and Phellinus baumii, on platelet aggregation (상황버섯, Phellinus linteus 계통과 Phellinus baumii의 항 혈소판 응집 효과)

  • Kang, Hee-Wan;Rhee, Man-Hee;Yeo, Woon-Hyung
    • Journal of Mushroom
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    • v.15 no.3
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    • pp.124-128
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    • 2017
  • The fruiting bodies of Sanghwang mushrooms, Phellinus linteus HN1009K (PLH) and Phellinus linteus (Korea Sanghwang, PLK), and Phellinus baumii (Jangsu Sangwhang, PB), were extracted with 70% methanol. The methanolic extracts of different concentrations ($5-200{\mu}g/ml$) showed inhibitory effects of 20-95% on plated aggregation induced by collagen (2.5 ug/ml), ADP (10 uM), and thrombin (0.1 U/ml). The PLH, PLK, and PB extracts (200 ug/ml) reduced ATP release from ADP-activated platelets by 50-60%. These results suggest that the methanolic extracts from Sanghwang mushrooms inhibit platelet aggregation.

Gene Expression Profiling in Osteoclast Precursors by Insulin Using Microarray Analysis

  • Kim, Hong Sung;Lee, Na Kyung
    • Molecules and Cells
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    • v.37 no.11
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    • pp.827-832
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    • 2014
  • The balance between bone formation by osteoblasts and destruction of mineralized bone matrix by osteoclasts is important for bone homeostasis. The increase of osteoclast differentiation by RANKL induces bone diseases such as osteoporosis. Recent studies have shown that insulin is one of main factors mediating the cross-talk between bone remodeling and energy metabolism. However, the systemic examination of insulin-induced differential gene expression profiles in osteoclasts has not been extensively studied. Here, we investigated the global effects of insulin on osteoclast precursors at the level of gene transcription by microarray analysis. The number of genes that were up-regulated by ${\geq}1.5$ fold after insulin treatment for 6 h, 12 h, or 24 h was 76, 73, and 39; and 96, 83, and 54 genes were down-regulated, respectively. The genes were classified by 20 biological processes or 24 molecular functions and the number of genes involved in 'development processes' and 'cell proliferation and differentiation' was 25 and 18, respectively, including Inhba, Socs, Plk3, Tnfsf4, and Plk1. The microarray results of these genes were verified by real-time RT-PCR analysis. We also compared the effects of insulin and RANKL on the expression of these genes. Most genes had a very similar pattern of expressions in insulin- and RANKL-treated cells. Interestingly, Tnfsf4 and Inhba genes were affected by insulin but not by RANKL. Taken together, these results suggest a potential role for insulin in osteoclast biology, thus contributing to the understanding of the pathogenesis and development of therapeutics for numerous bone and metabolic diseases.

Analyzing Place Location Knowledge Items of the Korean Geography Subject in the College Scholastic Ability Test: Focusing on Human (Economic) Geography (대학수학능력시험 한국지리 과목의 위치정보 문항 출제 경향 연구: 인문(경제)지리 문항을 중심으로)

  • Lee, Soyoung
    • Journal of the Economic Geographical Society of Korea
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    • v.24 no.1
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    • pp.29-51
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    • 2021
  • The present research explores the tendency of the items that require Place Location Knowledge (PLK) of the Korean Geography subject in the College Scholastic Ability Test. The major findings are as follows. First, the geographical regions of the items are spatially skewed, especially in the Yeongnam regions, which are tested more frequently compared to the others. Second, the fact-based items more concern with regionality such as geographic indication system and regional festivals. Third, the concept-based items can be divided into physical geography and human geography and there were four items related to economic geography. Fourth, students tend to find it challenging in the items asking PLK. The difficulty varies according to the type of items. The students find concept-based items which require high-order thinking more challenging. There is also differences identified between contents. For example, the section of physical geography, especially climatology-related, were considered the most challenging followed by those of economic geography. Finally, the differences in the rate of correct answer are associated with the scale of the regions covered in the items and students experienced more difficulty in the items asking more precise scale.

DNA Shuffling of aprE Genes to Increase Fibrinolytic Activity and Thermostability

  • Yao, Zhuang;Jeon, Hye Sung;Yoo, Ji Yeon;Kang, Yun Ji;Kim, Min Jae;Kim, Tae Jin;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.32 no.6
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    • pp.800-807
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    • 2022
  • Four aprE genes encoding alkaline serine proteases from B. subtilis strains were used as template genes for family gene shuffling. Shuffled genes obtained by DNase I digestion followed by consecutive primerless and regular PCR reactions were ligated with pHY300PLK, an E. coli-Bacillus shuttle vector. The ligation mixture was introduced into B. subtilis WB600 and one transformant (FSM4) showed higher fibrinolytic activity. DNA sequencing confirmed that the shuffled gene (aprEFSM4) consisted of DNA mostly originated from either aprEJS2 or aprE176 in addition to some DNA from either aprE3-5 or aprESJ4. Mature AprEFSM4 (275 amino acids) was different from mature AprEJS2 in 4 amino acids and mature AprE176 in 2 amino acids. aprEFSM4 was overexpressed in E. coli BL21 (DE3) by using pET26b(+) and recombinant AprEFSM4 was purified. The optimal temperature and pH of AprEFSM4 were similar to those of parental enzymes. However, AprEFM4 showed better thermostability and fibrinogen hydrolytic activity than the parental enzymes. The results indicated that DNA shuffling could be used to improve fibrinolytic enzymes from Bacillus sp. for industrial applications.