• Molecules and Cells
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• 제40권3호
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• pp.211-221
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• 2017

Transforming growth factor ${\beta}1$ $(TGF{\beta}1)/Smad4$ signaling plays a pivotal role in maintenance of the dynamic balance between bone formation and resorption. The microRNA miR-155 has been reported to exert a significant role in the differentiation of macrophage and dendritic cells. The goal of this study was to determine whether miR-155 regulates osteoclast differentiation through $TGF{\beta}1/Smad4$ signaling. Here, we present that $TGF{\beta}1$ elevated miR-155 levels during osteoclast differentiation through the stimulation of M-CSF and RANKL. Additionally, we found that silencing Smad4 attenuated the upregulation of miR-155 induced by $TGF{\beta}1$. The results of luciferase reporter experiments and ChIP assays demonstrated that $TGF{\beta}1$ promoted the binding of Smad4 to the miR-155 promoter at a site located in 454 bp from the transcription start site in vivo, further verifying that miR-155 is a transcriptional target of the $TGF{\beta}1/Smad4$ pathway. Subsequently, TRAP staining and qRT-PCR analysis revealed that silencing Smad4 impaired the $TGF{\beta}1$-mediated inhibition on osteoclast differentiation. Finally, we found that miR-155 may target SOCS1 and MITF to suppress osteoclast differentiation. Taken together, we provide the first evidence that $TGF{\beta}1/Smad4$ signaling affects osteoclast differentiation by regulation of miR-155 expression and the use of miR-155 as a potential therapeutic target for osteoclast-related diseases shows great promise.

• 한국축산식품학회지
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• 제43권4호
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• pp.703-711
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• 2023

As an initial study to elucidate the molecular mechanism of how probiotics modulate macrophage activity, we monitored mRNA expression patterns in peritoneal macrophages (PMs) treated with two different strains of probiotics. After treatment with either Weissella cibaria WIKIM28 or Latilactobacillus sakei WIKIM50, total RNAs from PMs were isolated and subjected into gene chip analyses. As controls, mRNAs from vehicle (phosphate-buffered saline, PBS)-treated PMs were also subjected to gene chip analysis. Compared to vehicle (PBS)-treated PMs, WIKIM28-treated and WIKIM50-treated PMs exhibited a total of 889 and 432 differentially expressed genes with expression differences of at least 4 folds, respectively. Compared to WIKIM28-treated PMs, WIKIM50-treated PMs showed 25 up-regulated genes and 21 down-regulated genes with expression differences of more than 2 folds. Interestingly, mRNA transcripts of M2 macrophage polarization marker such as anxa1, mafb, and sepp1 were increased in WIKIM50-treated PMs comparing to those in WIKIM28-treated PMs. Reversely, mRNA transcripts of M1 macrophage polarization marker such as hdac9, ptgs2, and socs3 were decreased in WIKIM50-treated PMs comparing to those in WIKIM28-treated PMs. In agreement with these observations, mRNA expression levels of tumor necrosis factor-α and interleukin-1α were significantly reduced in WIKIM50-treated macrophages compared to those in WIKIM28-treated macrophages. These results may indicate that probiotics can be classified as two different types depending on their ability to convert macrophages into M1 or M2 polarization.

• IMMUNE NETWORK
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• 제22권6호
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• pp.51.1-51.20
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• 2022

Trypanosoma cruzi, the etiological agent of Chagas disease, is an intracellular protozoan parasite, which is now present in most industrialized countries. About 40% of T. cruzi infected individuals will develop severe, incurable cardiovascular, gastrointestinal, or neurological disorders. The molecular mechanisms by which T. cruzi induces cardiopathogenesis remain to be determined. Previous studies showed that increased IL-6 expression in T. cruzi patients was associated with disease severity. IL-6 signaling was suggested to induce pro-inflammatory and pro-fibrotic responses, however, the role of this pathway during early infection remains to be elucidated. We reported that T. cruzi can dysregulate the expression of host PIWI-interacting RNAs (piRNAs) during early infection. Here, we aim to evaluate the dysregulation of IL-6 signaling and the piRNAs computationally predicted to target IL-6 molecules during early T. cruzi infection of primary human cardiac fibroblasts (PHCF). Using in silico analysis, we predict that piR_004506, piR_001356, and piR_017716 target IL6 and SOCS3 genes, respectively. We validated the piRNAs and target gene expression in T. cruzi challenged PHCF. Secreted IL-6, soluble gp-130, and sIL-6R in condition media were measured using a cytokine array and western blot analysis was used to measure pathway activation. We created a network of piRNAs, target genes, and genes within one degree of biological interaction. Our analysis revealed an inverse relationship between piRNA expression and the target transcripts during early infection, denoting the IL-6 pathway targeting piRNAs can be developed as potential therapeutics to mitigate T. cruzi cardiomyopathies.

• Molecules and Cells
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• 제40권4호
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• pp.254-261
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• 2017

Glioblastomas (GBM) are very difficult to treat and their aggressiveness is one of the main reasons for this as well as for the frequent recurrences. MicroRNAs post-transcriptionally regulate their target genes through interaction between their seed sequence and 3'UTR of the target mRNAs. We previously reported that miR-296-3p is regulated by neurofibromatosis 2 (NF2) and enhances the invasiveness of GBM cells via SOCS2/STAT3. In this study, we investigated whether miR-296-5p, which originates from the same precursor miRNA as miR-296-3p, can increase the invasiveness of GBM cells. It was observed that miR-296-5p potentiated the invasion of various GBM cells including LN229, T98G, and U87MG. Through bioinformatics approaches, two genes were identified as miR-296-5p targets: caspase-8 (CASP8) and nerve growth factor receptor (NGFR). From results obtained from Ago2 immunoprecipitation and luciferase assays, we found that miR-296-5p downregulates CASP8 and NGFR through direct interaction between seed sequence of the miRNA and 3'UTR of the target mRNA. Knockdown of CASP8 or NGFR also increased the invasive ability of GBM cells, indicating that CASP8 and NGFR are involved in potentiation of invasiveness by miR-296-5p. Consistent with our findings, CASP8 was downregulated in brain metastatic lung cancer cells, which have a high level of miR-296-5p, compared to parental cells, suggesting that miR-296-5p may be generally associated with the acquisition of invasiveness. Collectively, our results implicate miR-296-5p as a potential cause of invasiveness in cancer and suggest it as a promising therapeutic target for GBM.

• 정보보호학회논문지
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• 제30권3호
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• pp.465-479
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• 2020

국내·외 기업 및 기관들은 사이버위협으로부터 자신들의 IT 인프라를 안전하게 보호하기 위해 24시간/365일 모니터링 및 대응할 수 있는 보안관제센터를 활용하고 있다. 하지만, 현재 대부분의 보안관제센터는 전문 인력에 의한 수동분석과 텍스트 기반의 보안관제체계에 의존하는 태생적인 한계점을 안고 있다. 이러한 보안관제체계의 문제점들을 극복하기 위해 가시화 기술을 활용한 사이버위협 탐지·분석 연구가 활발하게 진행되고 있지만 이들 연구의 대부분은 보안관제 분야에 최적화되어 있지 않고, 많은 경우에 개별 기관에서만 활용할 수 있다는 제한이 따랐다. 따라서 본 논문에서는 보안관제 분야의 최종 목표인 실제 공격자 IP를 탐지할 수 있을 뿐만 아니라, 보안관제센터에서도 활용할 수 있는 새로운 가시화 방법론을 제안한다. 본 논문에서 제안하는 가시화 방법론의 핵심은 보안이벤트를 발생시킨 공격자(IP)의 행위정보를 실시간 및 추적(통계) 분석을 가능하게 하는 것이다. 제안된 가시화 방법론을 기반으로 개발된 시스템을 실제 보안관제센터에 성공적으로 적용하였으며, 실제 운영을 통해 다양한 공격자 IP를 탐지 및 분석하는데 성공함으로써 본 논문에서 제안한 가시화 방법론의 실용성 및 유효성을 검증했다.

• Clinical and Experimental Reproductive Medicine
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• 제32권3호
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• pp.269-277
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• 2005

Objectives: Previously, we sought to compile a list of genes expressed during early folliculogenesis by using cDNA microarray to investigate follicular gene expression and changes during primordialprimary follicle transition and development of secondary follicles (Yoon et al., 2005). Among those genes, a group of genes related to the cell size growth was characterized during the ovarian development in the present study. Methods: We determined ovarian expression pattern of six genes related to the cell size growth (cyr61, emp1, fhl1, socs2, wig1 and wisp1) and extended into CCN family (${\underline{c}}onnective$ tissue growth factor/${\underline{c}}ysteine$-rich 61/${\underline{n}}ephroblastoma$-overexpressed), ctgf, nov, wisp2, wisp3, including cyr61 and wisp1 genes. Expression of mRNA and protein according to the ovarian developmental stage was evaluated by in situ hybridization, and/or semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), and immunohistochemistry, respectively. Results: Among 6 genes related to the cell size growth, cyr61 and wisp1 mRNA was detected only in oocytes in the postnatal day5 mouse ovaries. cyr61 mRNA expression was limited to the nucleolus of oocytes, while wisp1 was expressed in the cytoplasm and nucleolus of oocytes, except nucleus. cyr61 mRNA expression, however, was found in granulosa cells from secondary follicles. The rest 4 genes in the cell size growth group were detected in oocytes, granulosa and theca cells. Cyr61 and Wisp1 proteins were expressed in the oocyte cytoplasm from primordial follicle stage. Especially, Cyr61 protein was detected in pre-granulosa cells, Wisp1 protein was not. By using RT-PCR, we evaluated and decided that Cyr61 protein is produced by their own mRNA in pre-granulosa cells that was not detected by in situ hybridization. cyr61 and wisp1 genes are happen to be the CCN family members. The other members of CCN family were also studied, but their expression was detected in oocytes, granulose and theca cells. Conclusions: We firstly characterized the ovarian expression of genes related to the cell size growth and CCN family according to the early folliculogenesis. Cyr61 protein expression in the pre-granulosa cells is profound in meaning. Further functional analysis for cyr61 in early folliculogenesis is under investigation.

• 한국측량학회지
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• 제40권3호
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• pp.159-168
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• 2022

지방자치단체는 생활SOC의 불균형을 해소하려는 노력을 추진되어야 하며, 생활SOC의 입지 현황, 최적입지 등에 관한 공간정보를 구축해야 한다. 이 연구는 생활SOC 관련 연구수요를 고려하여, 생활SOC의 접근성, 형평성, 그리고 최적입지를 분석하였다. 분석 시설은 생활거점시설로서, 공공도서관, 근린공원, 생활문화센터, 국공립어린이집으로 구성하였다. 분석지역은 경기도 수원시이며, 분석의 기준연도는 2020년이다. 접근성은 세생활권(200m×200m)에서 인구 1인당 접근성을 산출하였다. 형평성은 지니계수를 활용하여 동 간 접근성의 지역 간 격차를 확인하였다. 최적입지는 최대 커버링 입지문제를 활용하여 탐색하였다. 분석 결과, 국공립어린이집을 제외한 시설의 공간적 접근성은 지역 간 커다란 차이를 드러냈다. 접근성이 우수한 지역도 시설에 따라 커다란 편차를 보였다. 생활SOC의 지역 간 격차는 생활문화센터가 가장 컸으며, 뒤를 이어, 근린공원, 국공립어린이집, 공공도서관 순으로 나타났다. 공공도서관, 근린공원, 생활문화센터의 최적입지는 원도심에 집중적으로 분포하였으나, 국공립어린이집의 최적입지는 수원시 전역에서 나타났다. 이 연구의 결과는 추후 지방자치단체에서 생활SOC 공급계획을 수립함에 있어 핵심적인 계획내용이다. 따라서 후속연구는 이 연구의 분석구조와 결과를 유용하게 활용 가능할 것으로 기대한다.

• 생명과학회지
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• 제33권5호
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• pp.397-405
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• 2023

항산화제로서 산화적 손상의 방지에 중요한 역할을 하는 것으로 알려진 glutathione (GSH)의 면역 조절에 대한 연구는 현재까지 제대로 이루어지지 않았다. 본 연구에서 우리는 환원형 GSH인 luthione^®이 RAW 264.7 세포에서 면역 강화 효과가 있는지를 조사하였다. 유세포 분석 및 면역 형광 실험의 결과에 의하면, luthione은 대조군 세포에 비해 대식세포의 대표적인 기능인 식세포 활성을 luthione 처리 농도 의적으로 증가시키는 것으로 나타났다. 또한, cytokine array의 결과에 의하면, IL-5, IL-1β와 IL-27의 발현이 luthione이 처리된 세포에서 유의하게 증가하였다. 아울러 luthione에 의한 TNF-α 및 IL-1β의 생성 증가는 그들의 단백질 발현 증가를 통해 이루어졌으며, NO 및 PGE₂와 같은 면역 매개체 유리의 증가는 iNOS 및 COX-2의 발현 증가와 관련이 있었으며, 이는 M1 대식세포 분화 마커인 CD86 발현의 증가와 연관성이 있었다. 그리고 heatmap 분석을 통하여 SOCS1/3 매개 STAT/JAK 신호 전달 경로가 luthione에 의한 면역 조절 증가에 관여함을 확인하였다. 결론적으로, 우리의 결과는 luthione이 M1 macrophage polarization의 분자 조절자로 작용하여 면역 능력을 향상시킬 수 있음을 시사한다.

• Animal Bioscience
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• 제36권7호
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• pp.991-1002
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• 2023

Objective: This study aimed to elucidate the underlying gene regions responsible for productive, phenotypic or adaptive traits in different ecological types of Tibetan sheep and the discovery of important genes encoding valuable traits. Methods: We used whole-genome resequencing to explore the genetic relationships, phylogenetic tree, and population genetic structure analysis. In addition, we identified 28 representative Tibetan sheep single-nucleotide polymorphisms (SNPs) and genomic selective sweep regions with different traits in Tibetan sheep by fixation index (Fst) and the nucleotide diversity (θπ) ratio. Results: The genetic relationships analysis showed that each breed partitioned into its own clades and had close genetic relationships. We also identified many potential breed-specific selective sweep regions, including genes associated with hypoxic adaptability (MTOR, TRHDE, PDK1, PTPN9, TMTC2, SOX9, EPAS1, PDGFD, SOCS3, TGFBR3), coat color (MITF, MC1R, ERCC2, TCF25, ITCH, TYR, RALY, KIT), wool traits (COL4A2, ERC2, NOTCH2, ROCK1, FGF5, SOX9), and horn phenotypes (RXFP2). In particular, a horn-related gene, RXFP2, showed the four most significantly associated SNP loci (g. 29481646 A>G, g. 29469024 T>C, g. 29462010 C>T, g. 29461968 C>T) and haplotypes. Conclusion: This finding demonstrates the potential for genetic markers in future molecular breeding programs to improve selection for horn phenotypes. The results will facilitate the understanding of the genetic basis of production and adaptive unique traits in Chinese indigenous Tibetan sheep taxa and offer a reference for the molecular breeding of Tibetan sheep.

• Animal Bioscience
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• 제35권9호
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• pp.1340-1350
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• 2022

Objective: Luxi Black Head sheep (LBH) is the first crossbreed specialized for meat production and was developed by crossbreeding Black Head Dorper sheep (DP) and Small Tailed Han sheep (STH) in the farming areas of northern China. Research on the genomic variations and selection signatures of LBH caused by continuous artificial selection is of great significance for identifying the genetic mechanisms of important traits of sheep and for the continuous breeding of LBH. Methods: We explored the genetic relationships of LBH, DP, and several Mongolian sheep breeds by constructing phylogenetic tree, principal component analysis and linkage disequilibrium analysis. In addition, we analysed 29 whole genomes of sheep. The genome-wide selection signatures have been scanned with four methods: heterozygosity (H_P), fixation index (F_ST), cross-population extended haplotype homozygosity (XP-EHH) and the nucleotide diversity (𝜃_π) ratio. Results: The genetic relationships analysis showed that LBH appeared to be an independent cluster closer to DP. The candidate signatures of positive selection in sheep genome revealed candidate genes for developmental process (HoxA gene cluster, BCL2L11, TSHR), immunity (CXCL6, CXCL1, SKAP2, PTK6, MST1R), growth (PDGFD, FGF18, SRF, SOCS2), and reproduction (BCAS3, TRIM24, ASTL, FNDC3A). Moreover, two signalling pathways closely related to reproduction, the thyroid hormone signalling pathway and the oxytocin signalling pathway, were detected. Conclusion: The selective sweep analysis of LBH genome revealed candidate genes and signalling pathways associated with developmental process, immunity, growth, and reproduction. Our findings provide a valuable resource for sheep breeding and insight into the mechanisms of artificial selection.