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      Human microRNA-299-3p decreases invasive behavior of cancer cells by downregulation of Oct4 expression and causes apoptosis

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          Abstract

          Purpose

          Oct4 was reported to be one of the most important pluripotency transcription factors in the biology of stem cells including cancer stem cells, and progressed malignant cells. Here we report the investigation of gene expression control of Oct4 by selected human microRNAs and the physiological effect of Oct4 silencing in invasive cancer cells.

          Methods and results

          High throughput luciferase activity assay revealed the microRNA-299-3p to be the most effective in reducing gene expression of Oct4, which was confirmed by Western blot analysis and Oct4 promoter activity in a target luciferase assay. Furthermore, it could be demonstrated that downregulation of Oct4 by microRNAs-299-3p in breast cancer and fibrosarcoma cells lead to a decreased invasiveness in a microfluidic chip assay. Additionally, microRNA-299-3p causes apoptosis in cancer cells. Comparison with Oct4 specific siRNA transfection confirmed that this effect is primary due to the blockade of Oct4 expression.

          Conclusion

          The results suggest that microRNA-299-3p is an interesting target for potential clinical use. It may be able to decrease invasive behaviour of carcinoma cells; or even kill these cells by causing apoptosis.

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          Most cited references24

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          Physics and applications of microfluidics in biology.

          Fluid flow at the microscale exhibits unique phenomena that can be leveraged to fabricate devices and components capable of performing functions useful for biological studies. The physics of importance to microfluidics are reviewed. Common methods of fabricating microfluidic devices and systems are described. Components, including valves, mixers, and pumps, capable of controlling fluid flow by utilizing the physics of the microscale are presented. Techniques for sensing flow characteristics are described and examples of devices and systems that perform bioanalysis are presented. The focus of this review is microscale phenomena and the use of the physics of the scale to create devices and systems that provide functionality useful to the life sciences.
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            Vertebrate microRNA genes.

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              Both natural and designed micro RNAs can inhibit the expression of cognate mRNAs when expressed in human cells.

              Animal cells have recently been shown to express a range of approximately 22 nucleotide noncoding RNAs termed micro RNAs (miRNAs). Here, we show that the human mir-30 miRNA can be excised from irrelevant, endogenously transcribed mRNAs encompassing the predicted 71 nucleotide mir-30 precursor. Expression of the mir-30 miRNA specifically blocked the translation in human cells of an mRNA containing artificial mir-30 target sites. Similarly, designed miRNAs were also excised from transcripts encompassing artificial miRNA precursors and could inhibit the expression of mRNAs containing a complementary target site. These data indicate that novel miRNAs can be readily produced in vivo and can be designed to specifically inactivate the expression of selected target genes in human cells.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                20 April 2017
                2017
                : 12
                : 4
                : e0174912
                Affiliations
                [1 ]Experimentelle Nephrologie, Klinik für Innere Medizin III, Universitätsklinikum Jena, Jena, Germany
                [2 ]Abteilung Hämatologie und Internistische Onkologie, Klinik für Innere Medizin II, Universitätsklinikum Jena, Jena, Germany
                [3 ]Institut für Pharmazie und Molekulare Biotechnologie, Abteilung pharmazeutische Biologie, Universität Heidelberg, Heidelberg, Germany
                University of Hong Kong, HONG KONG
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: ARG RM.

                • Data curation: ARG XC JHC RM.

                • Formal analysis: ARG SR XC SW JT JHC RM.

                • Funding acquisition: RM.

                • Investigation: ARG XC JT RM.

                • Methodology: ARG JHC RM.

                • Project administration: ARG RM.

                • Resources: SW JHC RM.

                • Software: RM.

                • Supervision: SW RM.

                • Validation: ARG XC JT JHC RM.

                • Visualization: ARG XC.

                • Writing – original draft: ARG.

                • Writing – review & editing: SR XC SW JT JHC RM.

                Author information
                http://orcid.org/0000-0002-7176-8506
                Article
                PONE-D-17-09025
                10.1371/journal.pone.0174912
                5398498
                28426762
                2ec4f2dd-db5e-4383-9f1e-75b2f7042272
                © 2017 Göhring et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 7 March 2017
                : 17 March 2017
                Page count
                Figures: 7, Tables: 0, Pages: 15
                Funding
                Funded by: German Ministry for Education and Research BMBF
                Award ID: FKZ 0315389A and B
                Award Recipient :
                This work was supported by the Federal German Ministry of Education and Research (BMBF; http://www.bmbf.de/), within the Medical Systems Biology Project: Drug-iPS, FKZ 0315389A and B, and SysToxChip BMBF FKZ: 031A303A and E to RM and SW.
                Categories
                Research Article
                Biology and life sciences
                Genetics
                Gene expression
                Gene regulation
                MicroRNAs
                Biology and life sciences
                Biochemistry
                Nucleic acids
                RNA
                Non-coding RNA
                MicroRNAs
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cell Death
                Apoptosis
                Biology and Life Sciences
                Biochemistry
                Enzymology
                Enzymes
                Oxidoreductases
                Luciferase
                Biology and Life Sciences
                Biochemistry
                Proteins
                Enzymes
                Oxidoreductases
                Luciferase
                Medicine and Health Sciences
                Oncology
                Cancers and Neoplasms
                Breast Tumors
                Breast Cancer
                Biology and life sciences
                Genetics
                Gene expression
                Gene regulation
                Small interfering RNAs
                Biology and life sciences
                Biochemistry
                Nucleic acids
                RNA
                Non-coding RNA
                Small interfering RNAs
                Computer and Information Sciences
                Computing Methods
                Control Sequences
                Medicine and Health Sciences
                Oncology
                Cancers and Neoplasms
                Sarcomas
                Fibrosarcoma
                Biology and Life Sciences
                Cell Biology
                Cellular Types
                Animal Cells
                Stem Cells
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

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