Human exosomal placenta-specific miRNAs modulate expression of target genes in maternal immune cells during pregnancy
Ontology highlight
ABSTRACT: We investigated placenta-specific miRNA miR-517a (miR-517a-3p) functions in Jurkat cells. For this purpose, we identified candidate target mRNAs of the placenta-specific miRNAs using DNA microarray analysis. Transfection of Jurkat cells with miR-517a significantly downregulated 123 genes. Among the 123 genes identified, we searched for potential direct targets of miR-517a using the online software MicroCosm Targets. Seven genes, ALDH1B1, ANP32E, DHFR, FAT2, IGSF5, PRKG1, and RSPO3, had at least one potential miR-517a binding site in their 3M-bM-^@M-^Y-UTRs. Furthermore, we revealed that PRKG1, one of the seven genes, was a miR-517a target using an in vitro experimental validation system. We compared gene expression pattern of miR-517aM-bM-^@M-^Soverexpressing Jurkat cells with that of negative control.
Project description:The miR-29 family is an important player in the molecular pathophysiology of distinct types of cancer, with roles that seems to depend on cellular context. Reduced miR-29 levels are associated with more aggressive disease and its overexpression in cancer and leukemic cell lines inhibits proliferation. In contrast, its overexpression in hematopoietic progenitors, promotes leukemogenesis. We explored the potential roles of miR-29a in the molecular pathophysiology of T-cell acute lymphoblastic leukemia (T-ALL). As compared to normal T-cells, miR-29a levels are extremely reduced in T-ALL and in the Jurkat cell line. Microarrays analysis in Jurkat cells, following the introduction of synthetic miR-29a mimics, revealed the down-regulation of several predicted targets, including previously described targets (DNMT3a/b, CDK6, PXDN, MCL1, PIK3R1 and CXXC6), and novel targets with roles in active DNA demethylation, such as members of the ten-eleven-translocation (TET) family and TDG. Reduced miR-29a levels contribute to altered epigenetics, as its introduction in Jurkat cells, promotes the demethylation of the AHR gene (commonly methylated in T-ALL). In T-ALL patients, miR-29a levels are significantly associated with blast counts and disease free survival. Our results highlight the relevance of miR-29 in T-ALL physiopathology, and may help to clarify some contrasting findings reported in the literature. In order to identify potential miR-29a targets in T-cell lymphoblastic leukemia, Jurkat cells were electroporated with synthetic RNA molecules corresponding to miR-29a mimics (pre-miR) or inhibitors (anti-miR); and microarray profiles were compared to the profiles of Jurkat cells eletroporated with the corresponding negative controls. Transcripts with levels reduced following the introduction of the pre-miR-29a (as compared to pre-miR-Control), or transcripts acumulated folowing introduction of the anti-miR-29a inhibitor (as compared to anti-miR-Control), were considered potential targets; and were further compared to microRNA target prediction databases. Two paired samples were used for this analysis.
Project description:We generated a model for miR-34c-5p overexpression by transfecting Jurkat T cell line with a pcDNA3 vector containing the pre-miR-34c-5p precursor sequence and selecting for antibiotic resistance to isolate both pools and clones of stably transfected cells (J34c cells), in parallel with control cells transfected with the empty parental vector (JÃ cells). Both pools of stably transfected cells and the derived clonal lines expressed high levels of the miR.
Project description:miR-181 is often dysregulated in several types of cancer. This dataset can represent a further insight into gene expression changes and GO Terms associated with miR-181 over-expression. miR-181 was over-expressed in Jurkat cells by transduction with a lentiviral transgenic construct encoding for miR-181 under a PGK promoter. A cognate vector encoding for a control hairpin was used to generate control cell line. mRNA Microarray gene expression profiling of Jurkat cells tranduced with either a miR-181 sponge or control construct. 3 biological replicas have been performed. Note that the control samples are the same as those used in the related experiment E-MTAB-4588
Project description:We investigated placenta-specific miRNA miR-517a (miR-517a-3p) functions in Jurkat cells. For this purpose, we identified candidate target mRNAs of the placenta-specific miRNAs using DNA microarray analysis. Transfection of Jurkat cells with miR-517a significantly downregulated 123 genes. Among the 123 genes identified, we searched for potential direct targets of miR-517a using the online software MicroCosm Targets. Seven genes, ALDH1B1, ANP32E, DHFR, FAT2, IGSF5, PRKG1, and RSPO3, had at least one potential miR-517a binding site in their 3’-UTRs. Furthermore, we revealed that PRKG1, one of the seven genes, was a miR-517a target using an in vitro experimental validation system.
Project description:MiR-21 is an important suppressor of T-cell apoptosis that is also widely overexpressed in many types of cancers. The exact mechanisms related to the anti-apoptotic effect of miR-21 is not well understood. In this study, we applied AGO2 RNA Immunoprecipitation followed by gene expression profiling (RIP-Chip) in Jurkat cells to identify apoptosis-associated miR-21 target genes. We showed that expression of miR-21 rapidly increases upon αCD3/αCD28 activation of Jurkat cells. Inhibition of miR-21 resulted in reduced cell growth and induced apoptosis. Upon AGO2-RIP-Chip, we observed an overall increased enrichment of miR-21 target genes in the IP fraction of miR-21-overexpressing Jurkat cells as compared to the IP fraction of empty vector control cells. We noted a systematic decrease in transcript levels of predicted miR-21 target genes compared to EV control. We identified 72 genes that were 2-fold enriched in the AGO2-IP fraction of miR-21-overexpressing cells that contained a predicted miR-21 binding site. Of these, 71 were enriched 2-fold more in the miR-21-overexpressing cells as compared to EV Jurkat cells. The target gene for which the enrichment was most prominently increased upon miR-21 overexpression was the pro-apoptotic protein LATS1. Luciferase reporter assays confirmed direct targeting of the LATS1 3'UTR by miR-21. In line with the luciferase results, Western blot analysis revealed a decrease in LATS1 upon miR-21 inhibition. LATS1 qRT-PCR analysis in primary T-cells showed that LATS1 levels decrease upon T-cell stimulation while the miR-21 levels increase. Collectively, these data identify the miR-21 target LATS1 as a likely candidate whose inhibition contributes to the anti-apoptotic function of miR-21 in T-cells and perhaps also many types of cancers. Gene expression array on Jurkat cells overexpressing miR-21 and empty vector (EV).
Project description:miR-93 is often dysregulated in several tumor cell lines, including lymphoblastic leukemias. This dataset can represent a further insight into gene expression changes and GO Terms associated with miR-93 over-expression. miR-93 was over-expressed in Jurkat cells by transduction with a lentiviral transgenic construct encoding for miR-93 under a PGK promoter. A cognate vector encoding for a control hairpin was used to generate control cell line. mRNA Microarray gene expression profiling of Jurkat cells tranduced with either a miR-93 sponge or control construct. 3 biological replicas have been performed.
Project description:screenning the differentially expressed genes of Jurkat-FF3, Jurkat-miR146a, Jurkat-miR146a-sponge cell lines. To investigate the role of miR-146a on Jurkat T cells, we screened the differentially expressed genes of Jurkat-FF3 (as control), Jurkat-miR146a (as miR-146a overexpression cell line), Jurkat-miR146a-sponge (as miR-146a knock down cell line).
Project description:Screening the differetial expressed genes with Jurkat-146a(miR-146a over expression),Jurkat-sponge(miR-146a knockdown),Jurkat-FF3(vector control)
Project description:The biological significance of the down-regulation of miR-376c in HuCCT1 cells is unknown. We revealed that miR-376c significantly reduced EGF-dependent cell migration through its target GRB2 in HuCCT1 cells. To elucidate key molecules in EGF-dependent HuCCT1 migration, in which GRB2 upregulation is caused by abnormal suppression of miR-376c, we compared expression profiles of mRNAs affected by pre-miR-376c and siRNA targeting GRB2. We compared expression profiles of mRNAs affected by pre-miR-376c and siRNA targeting GRB2 in EGF-treated HuCCT1 cells.
Project description:miR-145 is downregulated in multiple cancers. The introduction of miR-145 could alleviate the tumor burden in the pancreatic cancer mouse model. However, how miR-145 mediates the tumor suppression is still an open question. In this study, we aimed to identify the targets of miR-145 using a SILAC approach.