IL6R is a target of miR-197 in human keratinocytes
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ABSTRACT:
PROVIDER: PRJNA650204 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR12365983.fastq.gz | Fastqsanger.gz | |||
| SRR12365984.fastq.gz | Fastqsanger.gz | |||
| SRR12365985.fastq.gz | Fastqsanger.gz | |||
| SRR12365986.fastq.gz | Fastqsanger.gz | |||
| SRR12365987.fastq.gz | Fastqsanger.gz |
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Project description:Background: Psoriasis is a chronic inflammatory disorder with cutaneous and systemic manifestations and substantial negative effects on patients' life quality. MicroRNAs (miRNAs) are post-transcriptional regulators of gene expression. Evidence suggests that miRNAs play a role in the pathogenesis of psoriasis. Previously studies, from others and by us, show-specific miRNAs that are differentially expressed between the psoriasis lesion and healthy skin. One of those miRNAs is miR-197-3p whose expression is down-regulated in the psoriatic lesions compared to normal or uninvolved psoriatic skin. In order to understand miR-197's role in psoriasis, we found that it could modulate IL-22 and IL-17 signaling. Objectives: We aimed to identify additional biochemical targets of miR-197 in psoriasis. Methods: We have chosen to apply a transcriptome-wide biochemical approach (PAR-CLIP) to search for new targets of miR-197 in keratinocytes. Results: in this work, using PAR-CLIP, we found an additional biochemical target of miR-197, one of the subunits of the receptor to IL-6. IL-6R is known to be up-regulated in psoriasis and even was considered as a possible therapeutic target. Conclusions: From the present data and our previous studies it appears that miR-197 is a major regulator of the interaction between immune system cells and keratinocytes.
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Project description:In general, microRNAs (miRNAs) regulate gene expression by repressing translation or promoting the degradation of their target genes. To strategically identify direct microR-197 (miR-197) target genes, we first performed mRNA microarray analyses of A549, PC14, and PC14CDDP cells that were transiently transfected with either pre-miR-197 or LNA-miR-197 and their controls. We observed that 1037 predicted target genes exhibited expression alterations consistently across the three different lung cancer cell lines.
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Project description:MicroRNAs are noncoding RNA species comprising 18–23 nucleotides that regulate host-virus interaction networks. Here, we showed that enterovirus A71 infection in human rhabdomyosarcoma (RD) involved miR-197 expression. miR-197 can regulate virus replication in the context of viral RNA synthesis via the transfection of its mimic into RD cells. We employed a mass spectrometry-based quantitative proteomic stable isotope labeling with amino acids in cell culture (SILAC) approach for the identification of miR-197 target genes by transfecting mimetic miR-197 into RD cells, and the differential expression of prospective target proteins was identified. A total of 1,822 genes were repeatedly and considerably downregulated in miR-197-transfected RD cells, 106 of which were predicted to have seed sites by TargetScan. Seven of the selected 8 genes potentially related to viral replication and immune response were confidently validated as direct miR-197 targets using a luciferase (untranslated region (3'-UTR)) reporter assay. The expression of three selected endogenous molecules (ITGAV, ETF1, and MAP2K1 (MEK1)) was significantly reduced when RD cells were transfected with an miR-197 mimic. Our results provide a database of miR-197 targets, which is potentially of interest in the area of viral pathogenesis and other research fields.
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Project description:First-line treatment for osteosarcoma includes chemotherapy and surgery. However, the five-year survival rate of refractory osteosarcoma remains unsatisfactory. Osteosarcoma cancer stem cells, possessing stemness and chemoresistance, are one of the critical causes for poor response to chemotherapy. Elucidating regulatory signaling pathways of osteosarcoma cancer stem cells may provide a rationale for improving regimens against chemoresistant osteosarcoma. Methotrexate (MTX)-resistant osteosarcoma cells were established. microRNA expression profiles were used for detecting differentially expressed microRNA in resistant clones and the parental cells. microRNA target databases were employed to predict potential microRNA and mRNA interaction. Flow cytometry was performed to measure stem cell marker Prominin-1 (CD133) positive cells. Im-munofluorescence staining was applied to detect CD133 expression. miR-197-3p mimic or an-ti-miR-197-3p stably transfected cells were used to generate xenograft models. In the study, we found miR-197-3p was increased in MTX-resistant cell lines. Overexpression of miR-197-3p en-hanced the expression of cancer stem cell markers CD133, Octamer-binding protein 4 (OCT4), Transcription factor SOX-2 (SOX2), and Homeobox protein NANOG (NANOG), as well as chemoresistance-associated genes ATP-dependent translocase ABCB1 (ABCB1) and Broad substrate specificity ATP-binding cassette transporter ABCG2 (ABCG2), whereas miR-197-3p knockdown inhibited stemness and recovered sensitivity to MTX. We also classified the tumor suppressor Speckle-type POZ protein-like (SPOPL) as a target of miR-197-3p. The miR-197-3p mutation that cannot combine SPOPL promoter regions was unable to sustain stemness or chemoresistance. Collectively, we discovered miR-197-3p conferred osteosarcoma stemness and chemotherapy resistance by targeting SPOPL, prompting promising therapeutic candidates for refractory oste-osarcoma treatment. miRNA qPCR assay
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