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Expression data of lentivirus sponge MicroRNA16 (miR16) downregulated RPMI 8226 and MMS1 multiple myeloma (MM) cell lines compared with control lentivirus sponge

ABSTRACT: Multiple myeloma is characterized by frequent chromosomal alterations. Deletion of chr 13, especially band 13q14, is commonly observed in early stages of MM, suggesting the presence of tumor suppressor genes (TSG) within this region. We sought to functionally validate the role of the microRNAs-15a/16 cluster, centered at the deleted region, as TSGs and delineate their downstream target genes in MM. Using âspongeâ lentiviral vectors to competitive stably inhibit mature microRNAs in vitro and in vivo, we have documented enhanced proliferative and invasive capacity of cells with stably inhibition of miR-16. Expression profiling analysis of miR-16-deficient cells identified a large number of downstream target genes. This loss-of-function system, which mimics the 13q chromosomal deletion, provides a valuable tool to investigate their function in MM pathogenesis and their potential use as therapeutic targets. We used microarrays to detail the global programme of gene expression underlying miR16 downregulation, simulating the 13q- cytogenetic changes related to MM in order to identified distinct classes of up-regulated genes during this process. MM cell lines were stably tranduced and sorted with miR16 lentiviruses expressing miR downregulating sponges or control sponges, and analyzed for GEP to asses possible miR16 target genes. Cells were lysed and the RNA extraction and hybridization was peformed on Affymetrix microarrays. We sought to obtain homogeneous populations of cells and therefore collected all at the same timepoint.

ORGANISM(S): Homo sapiens

SUBMITTER: Moshe Gat

PROVIDER: E-GEOD-24522 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Expression data of lentivirus sponge MicroRNA16 (miR16) downregulated RPMI 8226 and MMS1 multiple myeloma (MM) cell lines compared with control lentivirus sponge

Project description:Multiple myeloma is characterized by frequent chromosomal alterations. Deletion of chr 13, especially band 13q14, is commonly observed in early stages of MM, suggesting the presence of tumor suppressor genes (TSG) within this region. We sought to functionally validate the role of the microRNAs-15a/16 cluster, centered at the deleted region, as TSGs and delineate their downstream target genes in MM. Using “sponge” lentiviral vectors to competitive stably inhibit mature microRNAs in vitro and in vivo, we have documented enhanced proliferative and invasive capacity of cells with stably inhibition of miR-16. Expression profiling analysis of miR-16-deficient cells identified a large number of downstream target genes. This loss-of-function system, which mimics the 13q chromosomal deletion, provides a valuable tool to investigate their function in MM pathogenesis and their potential use as therapeutic targets.

2010-10-20 | GSE24522 | GEO

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Synthetic circular RNA functions as a miR-21 sponge to suppress gastric carcinoma cell proliferation

Project description:MicroRNA (miRNA) sponges containing miRNA complementary binding sites constitute a potentially useful strategy for miRNA-inhibition therapeutics in cancer patients. Recently, naturally occurring circular RNAs (circRNAs) have been revealed to function as efficient microRNA sponges. We hypothesized that synthetic circRNA sponges targeting oncomiRs could be constructed and used to achieve potentially therapeutic microRNA loss of function. In this study, linear RNA molecules containing five miR-21 binding sites were transcribed in vitro. After dephosphorylation by calf intestinal phosphatase and phosphorylation by T4 polynucleotide kinase, circRNA sponges were circularized using 5’-3’ end ligation by T4 RNA ligase 1. Synthetic circular sponge stability was assayed in the presence of RNase R or fetal bovine serum. Luciferase reporter and cell proliferation assays were performed to assess competitive inhibition of miR-21 activity by circRNA sponges in NCI-N87 gastric cancer cells. Tandem Mass Tag (TMT) labeling proteomics analysis and Western blotting were performed to delineate effects of circRNA sponges on miR-21 downstream targeted proteins. Our experiments revealed that artificial circRNA sponges can be synthesized using enzymatic ligation. These synthetic circRNA sponges are more resistant than their linear RNA counterparts to nuclease degradation in vitro. They effectively suppress the activity of miR-21 on its downstream protein targets, including the important cancer protein DAXX. Finally, they also inhibit gastric cancer cell proliferation. Our results suggest that synthetic circRNA sponges represent a rapid, effective, convenient strategy to achieve loss of miRNA function in vitro, with potential future therapeutic application in vivo.

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Expression data of lentivirus sponge Protein Arginine Methyltransferase 5 downregulated NCI-H929 cell line compared with control lentivirus sponge

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Quantitative Phosphoproteomics of whole-body sponge collapse

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