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As with a reduced expression in lymphoma relative to normal tissue only coincided partially e.g. for miR-218 and miR-200 family. Ten miRNAs that were present in the EBV-positive samples and thymus were not or almost undetectable in the EBV-negative tumours. Of note, miR449a and miR-449b were also not present in thymus. The other nine miRNAs, mainly members of the miR-200 family, showed reduced levels or comparable expression in the EBV-positive lymphomas as compared to thymus. Differentially BIBW2992 chemical information Expressed miRNAs in EBV-positive NK/T-cell vs. EBV-negative T-cell Lymphomas Furthermore, we compared the relative expression levels of the cellular miRNAs between EBV-positive and EBVnegative lymphomas. In the EBV-positive NK/T-cell lymphomas, 15 cellular miRNAs were up-regulated and 16 miRNAs were down-regulated relative to the EBV-negative ones. For the miRNAs shown in the diagram of Expression of Viral miRNAs in EBV-associated NK/T-cell Lymphomas Of note, we did not observe any EBV-miRNAs encoded by the BHRF1 cluster in accordance with previous results showing that these miRNAs are only found in cells that harbor the virus in a type III latency, such as a PTLD arising under immunosuppression but not, for instance, in NPC or stomach carcinoma. In contrast to the situation in NPC, where the EBV miRNAs accounted for 519% of the total miRNAs, the viral miRNAs in NK/T-cell lymphoma represented only 2.3% of the total miRNAs. The five most expressed EBV-miRNAs BART7, BART5, BART11-5p, BART1-5p and BART19-3p accounted for,50% of the viral encoded and for,1% of the total miRNAs. To confirm the absence of the EBV-encoded miRNAs from the BHRF1 cluster, the EBV-positive NK/T-cell lines HANKI and NK-YS were analysed by northern blotting. As shown in supporting Identification of New Cellular miRNAs Deregulated miRNAs in EBV-positive NK/T-cell and EBVnegative T-cell Lymphomas Relative to Normal Tissue We then compared the miRNA expression levels between the different small RNA libraries. For this analysis, we set a threshold of a 2-fold difference in expression and a representation of 0.1% in both small RNA libraries. First, we looked at the miRNA expression changes between the lymphomas and the normal MiRNA Profile of EBV-Positive NK/T-Cell Lymphoma 3 MiRNA Profile of EBV-Positive NK/T-Cell Lymphoma PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/22203538 analysis of the NK/T-cell lines did not yield any signal for the endogenous miRNA. Validation of miRNA Expression by Quantitative Realtime PCR We next tried to confirm the sequencing data by qRT-PCR. As outlined above, the nasal NK/T-cell lymphoma contain large necrotic areas; thus only limited amounts of material were available. We therefore could only analyse the levels of 4 upand 4 down-regulated miRNAs in the EBV-positive vs. -negative cases by PCR. We compared the expression of miR-205, -200c, 145, and -148a as well as miR-424, -142-3p, -181b, and -20b. but was found induced in EBV-positive vs. -negative lymphomas by PCR. The relative down-regulation of miR-424 for both lymphoma entities vs. thymus was confirmed, though. For all other miRNAs at least the same tendency of expression changes was detectable by PCR and by sequencing. Note that miR-205 was not detectable in the library of the EBV-negative lymphoma by sequencing; therefore, no relative values that compare the EBVnegative samples to the thymus and to the EBV-positive lymphomas can be displayed as log2. Nevertheless, the relative quantification of this miRNA by qRT-PCR confirmed the repression of miR

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Author: Adenosylmethionine- apoptosisinducer