Luciferase reporter assay for small-molecule inhibitors of MIR92b-3p function: Screening cyanopeptolins produced by Nostoc from the Baltic Sea

Brzuzan, Paweł; Mazur‐Marzec, Hanna; Florczyk, Maciej; Stefaniak, Filip; Fidor, Anna; Konkel, Robert; Woźny, Maciej

doi:10.1016/j.tiv.2020.104951

Cited by 9 publications

(8 citation statements)

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“…As the dysregulation of miRNA is linked to a number of diseases, including cancer, small molecules that interfere with miRNA are considered to be potential drug candidates [35,36]. In a previous study, a quantitative screening strategy to identify molecules that modulate miRNA 92b-3p expression in liver cells was developed [37]. The same screening strategy with Huh7 human hepatoma cell line stably transfected with a pmirGLO vector was used in current work to examine the effect of the three aeruginosamides on MIR92b-3p.…”

Section: Discussionmentioning

confidence: 99%

“…In this study, the reporter assays for small-molecule inhibitors of the MIR92b-3p function, previously designed to screen Nostoc peptides, were used [37]. The screen employs a Huh7 human hepatoma cell line (Cell Lines Service GmbH, CLS; Eppelheim, Germany) stably transfected with a pmirGLO vector containing dual-luciferase reporters.…”

Section: Luciferase Reporter Assaysmentioning

confidence: 99%

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Biological Activity and Stability of Aeruginosamides from Cyanobacteria

et al. 2022

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Aeruginosamides (AEGs) are classified as cyanobactins, ribosomally synthesized peptides with post-translational modifications. They have been identified in cyanobacteria of genera Microcystis, Oscillatoria, and Limnoraphis. In this work, the new data on the in vitro activities of three AEG variants, AEG A, AEG625 and AEG657, and their interactions with metabolic enzymes are reported. Two aeruginosamides, AEG625 and AEG657, decreased the viability of human breast cancer cell line T47D, but neither of the peptides was active against human liver cancer cell line Huh7. AEGs also did not change the expression of MIR92b-3p, but for AEG625, the induction of oxidative stress was observed. In the presence of a liver S9 fraction containing microsomal and cytosolic enzymes, AEG625 and AEG657 showed high stability. In the same assays, quick removal of AEG A was recorded. The peptides had mild activity against three cytochrome P450 enzymes, CYP2C9, CYP2D6 and CYP3A4, but only at the highest concentration used in the study (60 µM). The properties of AEGs, i.e., cytotoxic activity and in vitro interactions with important metabolic enzymes, form a good basis for further studies on their pharmacological potential.

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Section: Discussionmentioning

confidence: 99%

Section: Luciferase Reporter Assaysmentioning

confidence: 99%

Biological Activity and Stability of Aeruginosamides from Cyanobacteria

et al. 2022

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“…After 48 h of transfection, the luciferase activity (Firefly/Renilla) of each group was assessed using Luciferase Reporter Assay System (Promega). 22…”

Section: Methodsmentioning

confidence: 99%

“…After 48 h of transfection, the luciferase activity (Firefly/Renilla) of each group was assessed using Luciferase Reporter Assay System (Promega). 22 Statistical analysis SPSS 18.0 software (SPSS Inc., USA) was used for data analysis. Experimental data obtained from at least triplicate trails are shown as the mean ± standard deviation (SD).…”

Section: Luciferase Reporter Assaymentioning

confidence: 99%

MiR-362-5p inhibits cartilage repair in osteoarthritis via targeting plexin B1

Zhang

et al. 2022

J Orthop Surg (Hong Kong)

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Background and objectives Chondrogenesis of bone marrow mesenchymal stem cells (BMSCs) exerts great function during the pathogenesis of osteoarthritis (OA). Studies have reported the association of plexin B1 (PLXNB1) with OA pathogenesis. In this study, the upstream mechanism and function of PLXNB1 in this disease were explored. Methods Flow cytometry was applied to test BMSC characterization. Chondrogenic differentiation of BMSCs was evaluated by Alcian blue staining. The expression of PLXNB1, miR-362-5p, miR-501-5p, miR-1827, miR-500-5p was measured using RT-qPCR analysis. The protein levels of PLXNB1, Aggrecan, and Silent information regulator factor 2-related enzyme 1 (SIRT1) were determined by western blotting. Binding relationship between miR-362-5p and PLXNB1 was confirmed using bioinformatics analysis and luciferase reporter assay. The in vivo model of OA was established in Sprague-Dawley rats which received medial meniscus instability surgery. For histopathological examination, cartilage tissues in the knee joint of rats were stained with hematoxylin and eosin. Micro-CT analysis was employed to observe the changes of morphometric indices including average trabecular separation, average trabecular thickness, and bone volume fraction. Results BMSCs were identified to possess the characteristics of mesenchymal stem cells. PLXNB1 was observed to be highly expressed during chondrogenic differentiation of BMSCs and PLXNB1 overexpression promoted BMSC chondrogenic differentiation. Mechanically, PLXNB1 was targeted by miR-362-5p. In rescue assays, miR-362-5p reversed the effects of PLXNB1 on chondrogenic differentiation of BMSCs. In the in vivo experiments, upregulated PLXNB1 expression alleviated joint injury of OA rats. Additionally, overexpressed miR-362-5p and downregulated PLXNB1 expression levels were detected in OA rats. Conclusion MiR-362-5p promotes OA progression by suppressing PLXNB1.

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“…This tragedy marked a turning point in product testing, resulting in the development of systematic toxicity testing (Kim and Scialli, 2011). Hence, in the last decade in silico methods have been developing to assess both bioactivity and toxicity of known bioactive compounds and their synthetic analogs or novel compounds with known chemical structures (Yang et al, 2018;Liu et al, 2019;Brzuzan et al, 2020). In addition, the virtual screening technique called target fishing (Singh N. et al, 2020) can help recognizing the molecular target of the discovered marine natural products.…”

Section: Extraction Fractionation Isolation Structure Elucidationmentioning

confidence: 99%

The Essentials of Marine Biotechnology

Barbier²,

et al. 2021

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Coastal countries have traditionally relied on the existing marine resources (e.g., fishing, food, transport, recreation, and tourism) as well as tried to support new economic endeavors (ocean energy, desalination for water supply, and seabed mining). Modern societies and lifestyle resulted in an increased demand for dietary diversity, better health and well-being, new biomedicines, natural cosmeceuticals, environmental conservation, and sustainable energy sources. These societal needs stimulated the interest of researchers on the diverse and underexplored marine environments as promising and sustainable sources of biomolecules and biomass, and they are addressed by the emerging field of marine (blue) biotechnology. Blue biotechnology provides opportunities for a wide range of initiatives of commercial interest for the pharmaceutical, biomedical, cosmetic, nutraceutical, food, feed, agricultural, and related industries. This article synthesizes the essence, opportunities, responsibilities, and challenges encountered in marine biotechnology and outlines the attainment and valorization of directly derived or bio-inspired products from marine organisms. First, the concept of bioeconomy is introduced. Then, the diversity of marine bioresources including an overview of the most prominent marine organisms and their potential for biotechnological uses are described. This is followed by introducing methodologies for exploration of these resources and the main use case scenarios in energy, food and feed, agronomy, bioremediation and climate change, cosmeceuticals, bio-inspired materials, healthcare, and well-being sectors. The key aspects in the fields of legislation and funding are provided, with the emphasis on the importance of communication and stakeholder engagement at all levels of biotechnology development. Finally, vital overarching concepts, such as the quadruple helix and Responsible Research and Innovation principle are highlighted as important to follow within the marine biotechnology field. The authors of this review are collaborating under the European Commission-funded Cooperation in Science and Technology (COST) Action Ocean4Biotech – European transdisciplinary networking platform for marine biotechnology and focus the study on the European state of affairs.

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Luciferase reporter assay for small-molecule inhibitors of MIR92b-3p function: Screening cyanopeptolins produced by Nostoc from the Baltic Sea

Cited by 9 publications

References 41 publications

Biological Activity and Stability of Aeruginosamides from Cyanobacteria

Biological Activity and Stability of Aeruginosamides from Cyanobacteria

MiR-362-5p inhibits cartilage repair in osteoarthritis via targeting plexin B1

The Essentials of Marine Biotechnology

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