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1.
Oncogene ; 35(6): 670, 2016 02 11.
Article in English | MEDLINE | ID: mdl-32667144

ABSTRACT

At the request of the University of Luxembourg and following an external investigation, the Editor and Publisher have agreed to retract this paper owing to unreliable data.

2.
Hum Reprod ; 28(11): 3038-49, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23904466

ABSTRACT

STUDY QUESTION: What is the expression pattern of microRNAs (miRNAs) in human cumulus-oocyte complexes (COCs)? SUMMARY ANSWER: Several miRNAs are enriched in cumulus cells (CCs) or oocytes, and are predicted to target genes involved in biological functions of the COC. WHAT IS KNOWN ALREADY: The transcriptional profiles of human MII oocytes and the surrounding CCs are known. However, very limited data are available about post-transcriptional regulators, such as miRNAs. This is the first study focussing on the identification and quantification of small RNAs, including miRNAs, in human oocytes and CCs using a deep-sequencing approach. STUDY DESIGN, SIZE, DURATION: MII oocytes and CCs were collected from women who underwent IVF. PARTICIPANTS/MATERIALS, SETTING, METHODS: Using the Illumina/deep-sequencing technology, we analyzed the small RNAome of pooled MII oocytes (n = 24) and CC samples (n = 20). The mRNA targets of CC and MII oocyte miRNAs were identified using in silico prediction algorithms. Using oligonucleotide microarrays, genome-wide gene expression was studied in oocytes (10 pools of 19 ± 3 oocytes/each) and 10 individual CC samples. TaqMan miRNA assays were used to confirm the sequencing results in independent pools of MII oocytes (3 pools of 8 ± 3 oocytes/each) and CC samples (3 pools of 7 ± 3 CCs/each). The functional role of one miRNA, MIR23a, was assessed in primary cultures of human CCs. MAIN RESULTS AND THE ROLE OF CHANCE: Deep sequencing of small RNAs yielded more than 1 million raw reads. By mapping reads with a single location to the human genome, known miRNAs that were abundant in MII oocytes (MIR184, MIR100 and MIR10A) or CCs (MIR29a, MIR30d, MIR21, MIR93, MIR320a, MIR125a and the LET7 family) were identified. Predicted target genes of the oocyte miRNAs were associated with the regulation of transcription and cell cycle, whereas genes targeted by CC miRNAs were involved in extracellular matrix and apoptosis. Comparison of the predicted miRNA target genes and mRNA microarray data resulted in a list of 224 target genes that were differentially expressed in MII oocytes and CCs, including PTGS2, CTGF and BMPR1B that are important for cumulus-oocyte communication. Functional analysis using primary CC cultures revealed that BCL2 and CYP19A1 mRNA levels were decreased upon MIR23a overexpression. LIMITATIONS, REASONS FOR CAUTION: Only known miRNAs were investigated in the present study on COCs. Moreover, the source of the material is MII oocytes that failed to fertilize. WIDER IMPLICATIONS OF THE FINDINGS: The present findings suggest that miRNA could play a role in the regulation of the oocyte and CC crosstalk. STUDY FUNDING/COMPETING INTEREST(S): This work was partially supported by a grant from Ferring Pharmaceuticals. The authors of the study have no conflict of interest to report. TRIAL REGISTRATION NUMBER: Not applicable.


Subject(s)
Gene Expression Regulation , MicroRNAs/metabolism , Cumulus Cells/metabolism , Cumulus Cells/physiology , Female , Gene Expression Profiling , Humans , MicroRNAs/physiology , Oocytes/metabolism , Oocytes/physiology , Sequence Analysis, RNA
3.
Oncogene ; 29(31): 4436-48, 2010 08 05.
Article in English | MEDLINE | ID: mdl-20543867

ABSTRACT

Epithelial to mesenchymal transition (EMT) is a key step toward metastasis. MCF7 breast cancer cells conditionally expressing the EMT master regulator SNAI1 were used to identify early expressed microRNAs (miRNAs) and their targets that may contribute to the EMT process. Potential targets of miRNAs were identified by matching lists of in silico predicted targets and of inversely expressed mRNAs. MiRNAs were ranked based on the number of predicted hits, highlighting miR-661, a miRNA with so far no reported role in EMT. MiR-661 was found required for efficient invasion of breast cancer cells by destabilizing two of its predicted mRNA targets, the cell-cell adhesion protein Nectin-1 and the lipid transferase StarD10, resulting, in turn, in the downregulation of epithelial markers. Reexpression of Nectin-1 or StarD10 lacking the 3'-untranslated region counteracted SNAI1-induced invasion. Importantly, analysis of public transcriptomic data from a cohort of 295 well-characterized breast tumor specimen revealed that expression of StarD10 is highly associated with markers of luminal subtypes whereas its loss negatively correlated with the EMT-related, basal-like subtype. Collectively, our non-a priori approach revealed a nonpredicted link between SNAI1-triggered EMT and the down-regulation of Nectin-1 and StarD10 through the up-regulation of miR-661, which may contribute to the invasion of breast cancer cells and poor disease outcome.


Subject(s)
Breast Neoplasms/pathology , Cell Adhesion Molecules/genetics , Cell Dedifferentiation/genetics , MicroRNAs/genetics , Phosphoproteins/genetics , Transcription Factors/physiology , Breast Neoplasms/genetics , Breast Neoplasms/metabolism , Cell Adhesion Molecules/metabolism , Cell Dedifferentiation/drug effects , Cell Dedifferentiation/physiology , Epithelial Cells/metabolism , Epithelial Cells/physiology , Female , Gene Expression/physiology , Gene Expression Profiling , Gene Expression Regulation, Neoplastic/drug effects , Gene Expression Regulation, Neoplastic/physiology , Humans , Mesenchymal Stem Cells/metabolism , Mesenchymal Stem Cells/physiology , MicroRNAs/metabolism , MicroRNAs/physiology , Nectins , Neoplasm Invasiveness , Oligonucleotide Array Sequence Analysis , Phosphoproteins/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA, Small Interfering/pharmacology , Snail Family Transcription Factors , Transcription Factors/genetics , Transcription Factors/metabolism , Tumor Cells, Cultured , Validation Studies as Topic
5.
J Virol ; 74(9): 4064-73, 2000 May.
Article in English | MEDLINE | ID: mdl-10756018

ABSTRACT

Foamy viruses (FVs) are complex retroviruses which have been isolated from different animal species including nonhuman primates, cattle, and cats. Here, we report the isolation and characterization of a new FV isolated from blood samples of horses. Similar to other FVs, the equine foamy virus (EFV) exhibits a highly characteristic ultrastructure and induces syncytium formation and subsequent cell lysis on a large number of cell lines. Molecular cloning of EFV reveals that the general organization is that of other known FVs, whereas sequence similarity with its bovine FV counterpart is only 40%. Interestingly, EFV buds exclusively from the plasma membrane and not from the endoplasmic reticulum (ER), as previously shown for other FVs. The absence of the ER retrieval dilysine motif in EFV Env is likely responsible for this unexpected sorting pathway.


Subject(s)
Horse Diseases/virology , Retroviridae Infections/veterinary , Spumavirus/isolation & purification , Amino Acid Sequence , Animals , Antibodies, Viral/blood , Antibodies, Viral/immunology , Antigens, Viral/immunology , Base Sequence , Blotting, Southern , COS Cells , Cats , Cattle , Cloning, Molecular , Cricetinae , DNA, Viral/analysis , Genes, env , Genes, gag , Genes, pol , Horse Diseases/blood , Horse Diseases/immunology , Horses , Humans , Molecular Sequence Data , Proviruses/genetics , Rabbits , Retroviridae Infections/blood , Retroviridae Infections/immunology , Retroviridae Infections/virology , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Spumavirus/genetics , Spumavirus/immunology , Terminal Repeat Sequences , Transcriptional Activation , Tumor Cells, Cultured
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