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MicroRNA-3162-5p-Mediated Crosstalk between Kallikrein Family Members Including Prostate-Specific Antigen in Prostate Cancer
No full textBACKGROUND:
MicroRNAs mediate biological processes through preferential binding to the 3′ untranslated region (3′ UTR) of target genes. Studies have shown their association with prostate cancer (PCa) risk through single-nucleotide polymorphisms (SNPs), known as miRSNPs. In a European cohort, 22 PCa risk-associated miRSNPs have been identified. The most significant miRSNP in the 3′ UTR of Kallikrein-related peptidase 3 (KLK3) created a binding site for miR-3162-5p. Here we investigated the miR-3162-5p–KLK interaction and the clinical implication of miR-3162-5p in PCa.
METHODS:
We tested the role of miR-3162-5p in PCa etiology using IncuCyte live-cell imaging and anchorage-independent growth assays. The effect of miR-3162-5p on KLK and androgen receptor (AR) expression was measured by RT-quantitative (q)PCR and target pulldown assays. KLK3 proteolytic activity was determined by DELFIA® immunoassay. Mass spectrometry identified pathways affected by miR-3162-5p. miR-3162-5p expression was measured in clinical samples using RT-qPCR.
RESULTS:
miR-3162-5p affected proliferation, migration, and colony formation of LNCaP cells by regulating the expression of KLK2–4 and AR by direct targeting. KLK3 protein expression was regulated by miR-3162-5p consistent with lower KLK3 proteolytic activity observed in LNCaP-conditioned media. KLK/AR pulldown and mass spectrometry analysis showed a potential role of miR-3162-5p in metabolic pathways via KLK/AR and additional targets. Increased miR-3162-5p expression was observed in prostate tumor tissues with higher Gleason grade.
CONCLUSIONS:
Our study provides an insight into possible involvement of miR-3162-5p in PCa etiology by targeting KLKs and AR. It highlights clinical utility of miR-3162-5p and its interactive axis as a new class of biomarkers and therapeutic targets for PCa.No Full Tex
Santa Fe (ATSF) 3162
No full textA photograph print showing Santa Fe 3162, 2-8-0 (BLW, class 3160), South Shawnee, OK
β-catenin is a direct target of miR-3162-3p.
No full text(A) The three mRNA transcript variants of β-catenin possess different 3’-UTRs, which contain either one or two miR-3162-3p pairing sites. Site 2 is a common pairing site for all three transcript variants. Transcript variants 2 and 3 have only one pairing site (site 2) and transcript variant 1 has two pairing sites, sites 1 and 2. (B) Potential matching target sites of miR-3162-3p in the 3’-UTR of β-catenin mRNA transcript and sequence information of the wild type and mutant luciferase report constructs. Two fragments which comprise miR-3162-3p target sites on the 3′-UTR of β-catenin mRNA transcript were separately inserted into the luciferase gene of the pmirGLO vector at the NheI and SalI restriction sites. (C) The physical interaction between miR-3162-3p and the predicted target sites on β-catenin 3′-UTR in A549 cells was detected by luciferase reporter assay. Each wild type reporter construct was co-transfected with mimic-miR-3162-3p, syn-miR control, anti-miR-3162-3p or anti-miR control. The mutant reporter construct was co-transfected with mimic-miR-3162-3p or syn-miR control. Luciferase activity was measured at 24 h after transfection. Activity of the 3’-UTR reporter constructs was normalized to Renilla. Results are shown as the mean ± SD (n = 3). *p < 0.05 vs. syn-miR control.</p
MicroRNA-3162-5p-mediated crosstalk between kallikrein family members including prostate-specific antigen in prostate cancer
No full textBACKGROUND: MicroRNAs mediate biological processes through preferential binding to the 3′ untranslated region (3 UTR) of target genes. Studies have shown their association with prostate cancer (PCa) risk through singlenucleotide polymorphisms (SNPs), known as miRSNPs. In a European cohort, 22 PCa risk-associated miRSNPs have been identified. The most significant miRSNP in the 3′ UTR of Kallikrein-related peptidase 3 (KLK3) created a binding site for miR-3162-5p. Here we investigated the miR-3162-5p-KLK interaction and the clinical implication of miR-3162-5p in PCa. METHODS: We tested the role of miR-3162-5p in PCa etiology using IncuCyte live-cell imaging and anchorageindependent growth assays. The effect of miR-3162-5p on KLK and androgen receptor (AR) expression was measured by RT-quantitative (q)PCR and target pulldown assays. KLK3 proteolytic activity was determined by DELFIA immunoassay. Mass spectrometry identified pathways affected by miR-3162-5p. miR-3162-5p expression was measured in clinical samples using RT-qPCR. RESULTS: miR-3162-5p affected proliferation, migration, and colony formation of LNCaP cells by regulating the expression of KLK2-4 and AR by direct targeting. KLK3 protein expression was regulated by miR-3162-5p consistent with lower KLK3 proteolytic activity observed in LNCaP-conditioned media. KLK/AR pulldown and mass spectrometry analysis showed a potential role of miR-3162-5p in metabolic pathways via KLK/AR and additional targets. Increased miR-3162-5p expression was observed in prostate tumor tissues with higher Gleason grade. CONCLUSIONS: Our study provides an insight into possible involvement of miR-3162-5p in PCa etiology by targeting KLKs and AR. It highlights clinical utility of miR- 3162-5p and its interactive axis as a new class of biomarkers and therapeutic targets for PCa
MiR-3162-3p Is a Novel MicroRNA That Exacerbates Asthma by Regulating β-Catenin.
No full textAsthma is a common chronic respiratory disease. In a previous study, we found several circulating microRNA signatures associated with childhood asthma and selected miR-3162-3p for subsequent studies. Since the target proteins and underlying molecular mechanisms of miR-3162-3p in asthma etiopathogenesis are not well characterized, we designed this study to clarify its role. We employed bioinformatics and quantitative PCR methods as a first step to determine the target of miR-3162-3p, and we elucidated β-catenin. Luciferase assays and western blot analysis confirmed β-catenin as a direct target of miR-3162-3p as the 3'-untranslated region of β-catenin mRNA possesses a specific miR-3162-3p pairing site. The correlation between the expression levels of miR-3162-3p and β-catenin is confirmed by quantitative PCR and western blot studies in A549, Beas-2B and H1299 cell lines and OVA-induced asthma mouse model. Of note, upregulation of the endogenous miR-3162-3p level is concomitant with the reduction of β-catenin mRNA and protein expression levels. MiR-3162-3p antagomir treatment antagonizes the endogenous miR-3162-3p and effectively rescues the attenuation of endogenous β-catenin in OVA-induced asthmatic mice, which alleviates airway hyperresponsiveness and ameliorates airway inflammation. Collectively, our findings suggest a novel relationship between miR-3162-3p and β-catenin and clarify their mechanistic role in asthma etiopathogenesis
Block Card 3162 Radford Avenue
No full textThis image was produced by the Auditor's Office in Lucas County, Ohio for tax assessment purposes. Associated dates are approximate. Descriptive terms related to this photograph include: Ranch houses | 3162 Radford Avenue (Toledo, Ohio) | Dwelling | Replat of Vanderbilt Terrace (Toledo, Ohio) | South Toledo Area (Toledo, Ohio
Block Card 3162 Rocksberry Avenue
No full textThis image was produced by the Auditor's Office in Lucas County, Ohio for tax assessment purposes. Associated dates are approximate. Descriptive terms related to this photograph include: Garrison Colonial Style | 3162 Rocksberry Avenue (Toledo, Ohio) | Dwelling | Penrock Place Addition (Toledo, Ohio) | South Toledo Area (Toledo, Ohio
Block Card 3162 Strathmoor Avenue
No full textThis image was produced by the Auditor's Office in Lucas County, Ohio for tax assessment purposes. Associated dates are approximate. Descriptive terms related to this photograph include: Garrison Colonial Style | 3162 Strathmoor Avenue (Toledo, Ohio) | Dwelling | Heather Downs Addition (Toledo, Ohio) | South Toledo Area (Toledo, Ohio
Block Card 3162 Glanzman Road
No full textThis image was produced by the Auditor's Office in Lucas County, Ohio for tax assessment purposes. Associated dates are approximate. Descriptive terms related to this photograph include: 3162 Glanzman Road (Toledo, Ohio) | Dwelling | Heather Downs Addition (Toledo, Ohio) | South Toledo Area (Toledo, Ohio) | American four and a half squar
Block Card 3162 Glencairn Avenue
No full textThis image was produced by the Auditor's Office in Lucas County, Ohio for tax assessment purposes. Associated dates are approximate. Descriptive terms related to this photograph include: Ranch houses | Dwelling | Heather Downs Addition (Toledo, Ohio) | South Toledo Area (Toledo, Ohio) | 3162 Glencairn Avenue (Toledo, Ohio
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