Possible involvement of miRNAs in tropism of Parvovirus B19
| Autor: | Ehsan Arefian, Masoud Soleimani, Azadeh Anbarlou, Amir Atashi, Mahshid AkhavanRahnama, Giorgio Gallinella |
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| Přispěvatelé: | Anbarlou, Azadeh, Akhavanrahnama, Mahshid, Atashi, Amir, Soleimani, Masoud, Arefian, Ehsan, Gallinella, Giorgio |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
HEK-293 Parvovirus B19 Tropism Parvoviridae Infections 03 medical and health sciences Genetic microRNA Parvovirus B19 Human Genetics Humans Luciferase RNA Messenger Molecular Biology Drosha Erythroid Precursor Cells biology HEK 293 cells MCF7 Computational Biology Erythroid Progenitor General Medicine Transfection Virology Cell biology MicroRNAs HEK293 Cells 030104 developmental biology Gene Expression Regulation Cell culture MCF-7 Cells biology.protein Capsid Proteins Dicer |
| Popis: | Human Parvovirus B19 (PVB19) is one of the most important pathogens that targets erythroid lineage. Many factors were mentioned for restriction to erythroid progenitor cells (EPCs). Previous studies showed that in non-permissive cells VP1 and VP2 (structural proteins) mRNAs were detected but could not translate to proteins. A bioinformatics study showed that this inhibition might be due to specific microRNAs (miRNAs) present in non-permissive cells but not in permissive EPCs. To confirm the hypothesis, we evaluated the effect of miRNAs on VP expression. CD34(+) HSCs were separated from cord blood. Then, CD34(+) cells were treated with differentiation medium to obtain CD36(+) EPCs. To evaluate the effect of miRNAs on VP expression in MCF7 and HEK-293 cell lines (non-permissive cells) and CD36(+) EPCs, dual luciferase assay was performed in presence of shRNAs against Dicer and Drosha to disrupt miRNA biogenesis. QRT-PCR was performed to check down-regulation of Dicer and Drosha after transfection. All measurements were done in triplicate. Data means were compared using one-way ANOVAs. MicroRNA prediction was done by the online microRNA prediction tools. No significant difference was shown in luciferase activity of CD36(+) EPCs after co-transfection with shRNAs, while it was significant in non-permissive cells. Our study revealed that miRNAs may be involved in inhibition of VP expression in non-permissive cells, although further studies are required to demonstrate which miRNAs exactly are involved in regulation of PVB19 replication. |
| Databáze: | OpenAIRE |
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