• BMB Reports
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• 제51권11호
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• pp.602-607
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• 2018

Aberrant expression of microRNAs (miRNAs) plays important roles in carcinogenesis and tumor progression. However, the expression and biological role of miR-301b in triple-negative breast cancer (TNBC) remains unclear. Here we aimed to evaluate the roles and mechanisms of miR-301b in TNBC cells. miR-301b expression was assessed in TNBC specimens and cell lines by quantitative Real-Time PCR (qRT-PCR). TNBC cells were transfected with miR-301b mimics, inhibitors or Cylindromatosis (CYLD) small interfering RNA (siRNA) using Lipofectamine 2000. The functional roles of miR-301b were determined by cell proliferation, colony formation, and apoptosis assays. Western blots and qRT-PCR were used to measure the expression of mRNAs and proteins in the cells. We found that miR-301b was upregulated in TNBC specimens and cell lines. Overexpression of miR-301b promoted cell proliferation in TNBC cells, while inhibited the apoptosis induced by 5-FU. CYLD was downregulated by miR-301b at both mRNA and protein levels in TNBC cells. Dual-luciferase report assay confirmed that miR-301b downregulated CYLD by direct interaction with the 3'-untranslated region(3'-UTR) of CYLD mRNA. $NF-{\kappa}B$ activation was mechanistically associated with miR-301b-mediated downregulation of CYLD. However, inhibition of miR-301b reversed all the effects of miR-301b. In conclusion, miR-301b plays an oncogenic role in TNBC possibly by downregulating CYLD and subsequently activating $NF-{\kappa}B$ p65, and this may provide a novel therapeutic approach for TNBC.

• Journal of Gastric Cancer
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• 제19권3호
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• pp.301-314
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• 2019

Purpose: Peritoneal carcinomatosis in gastric cancer (GC) patients results in extremely poor prognosis. Malignant ascites samples are the most appropriate biological material to use to evaluate biomarkers for peritoneal carcinomatosis. This study identified exosomal MicroRNAs (miRNAs) differently expressed between benign liver cirrhosis-associated ascites (LC-ascites) and malignant gastric cancer-associated ascites (GC-ascites), and validated their role as diagnostic biomarkers for GC-ascites. Materials and Methods: Total RNA was extracted from exosomes isolated from 165 ascites samples (73 LC-ascites and 92 GC-ascites). Initially, microarrays were used to screen the expression levels of 2,006 miRNAs in the discovery cohort (n=22). Subsequently, quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) analyses were performed to validate the expression levels of selected exosomal miRNAs in the training (n=70) and validation (n=73) cohorts. Furthermore, carcinoembryonic antigen (CEA) levels were determined in ascites samples. Results: The miR-574-3p, miR-181b-5p, miR-4481, and miR-181d were significantly downregulated in the GC-ascites samples compared to the LC-ascites samples, and miR-181b-5p showed the best diagnostic performance for GC-ascites (area under the curve [AUC]=0.798 and 0.846 for the training and validation cohorts, respectively). The diagnostic performance of CEA for GC-ascites was improved by the combined analysis of miR-181b-5p and CEA (AUC=0.981 and 0.946 for the training and validation cohorts, respectively). Conclusions: We identified exosomal miRNAs capable of distinguishing between non-malignant and GC-ascites, showing that the combined use of miR-181b-5p and CEA could improve diagnosis.

• Molecules and Cells
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• 제38권10호
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• pp.895-903
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• 2015

Non-coding microRNAs (miRNAs) regulate the translation of target messenger RNAs (mRNAs) involved in the growth and development of a variety of cells, including primordial germ cells (PGCs) which play an essential role in germ cell development. However, the target mRNAs and the regulatory networks influenced by miRNAs in PGCs remain unclear. Here, we demonstrate a novel miRNAs control PGC development through targeting mRNAs involved in various cellular pathways. We reveal the PGC-enriched expression patterns of nine miRNAs, including miR-10b, -18a, -93, -106b, -126-3p, -127, -181a, -181b, and -301, using miRNA expression analysis along with mRNA microarray analysis in PGCs, embryonic gonads, and postnatal testes. These miRNAs are highly expressed in PGCs, as demonstrated by Northern blotting, miRNA in situ hybridization assay, and miRNA qPCR analysis. This integrative study utilizing mRNA microarray analysis and miRNA target prediction demonstrates the regulatory networks through which these miRNAs regulate their potential target genes during PGC development. The elucidated networks of miRNAs disclose a coordinated molecular mechanism by which these miRNAs regulate distinct cellular pathways in PGCs that determine germ cell development.

• 한국해양학회지:바다
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• 제19권4호
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• pp.287-301
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• 2014

냉수대 발생 전 후의 해양 환경과 식물플랑크톤 군집 구조 및 크기를 파악을 위해 여름철 빈번하게 냉수대가 발생되는 동해 남부 해역(울산 정자~부산 일광) 18개 정점에서 2013년 5월부터 8월까지 냉수대 발생 환경 및 식물플랑크톤 군집 구조를 조사하였다. 냉수대는 7월과 8월에 연안 정점(A1, B1, C1)에서 발생하였고, 표층에서 저온 고염의 특성을 보였다. 이 시기에 영양염은 수온과 유의한 음의 상관관계(DIP, r=-0.218, p<0.01; DIN, r=-0.306, p<0.01; silicate, r=-0.274, p<0.01)를 보여, 찬 해수가 분포하는 표층에 영양염이 풍부함을 알 수 있었다. 출현한 식물플랑크톤은 총 186종이었고, 현존량은 5월(C1, $726{\times}10^3cells\;L^{-1}$)과 7월(A1, $539{\times}10^3cells\;L^{-1}$)에 높았다. 또한, 연안 정점에서 총 chl. a 와 소형플랑크톤 chl. a ($&gt;20{\mu}m$)의 농도가 냉수대 발생 시기인 7, 8월에 뚜렷하게 증가한 반면, 수온약층이 형성된 6월에는 현저하게 낮았다. 6월의 우점종은 Pseudo-nitzschia spp.이었고, 그 세포 크기는 $309{\mu}m^3$로 다른 시기의 식물플랑크톤의 1/10 수준에 머무는 작은 크기였다. 이러한 결과는 7월과 8월에 총 chl. a의 증가와 식물플랑크톤의 크기 증가가 냉수대 발생 시에 표층으로 공급된 영양염의 영향임을 시사한다. 이러한 해양 환경 특성과 식물플랑크톤 출현양상은 연안 정점에서 뚜렷하게 보였고, 외측 정점에서는 관찰되지 않았다. 이 연구에서는 동해 남부 해역에서 냉수대가 발생하면 식물플랑크톤의 현존량 증가와 더불어 비교적 크기가 큰 식물플랑크톤의 출현 빈도가 증가하는 현상을 확인하였다. 결과적으로 하계에도 불구하고 동해 남부 연안 해역에서 냉수대 발생에 따른 영양염 공급은 식물플랑크톤 군집조성과 현존량에 상당한 영향을 미치는 것으로 판단된다.