Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Identification of latent tuberculosis infection-related microRNAs in human U937 macrophages expressing Mycobacterium tuberculosis Hsp16.3

ABSTRACT: Latent tuberculosis infection (LTBI) relies on a homeostasis of macrophages and Mycobacterium tuberculosis (Mtb). The small heat shock protein, Mtb Hsp16.3 (also known as latency-associated antigen), plays an important role in Mtb persistence within macrophages. However, the mechanism of LTBI remains elusive. The aim of this study was to delineate LTBI-related miRNA expression in U937 macrophages expressing Mtb Hsp16.3 protein. This study intends to explore the potential function of miRNAs in the interaction of macrophages with Mtb Hsp16.3 and provide insights for investigating the role of macrophage homeostasis in LTBI. U937 macrophages were infected with an integrase-deficient Lentivirus vector to transiently express Mtb Hsp16.3, and green fluorescent protein (GFP) as a control. We used a microRNA (miRNA) microarray chip containing more than 1000 probes to identify the significant differentially expressed miRNAs in the infected U937 cells, and employed real-time quantitative polymerase chain reaction (qRT-PCR) for validation. Furthermore, we confirmed these candidate LTBI-related miRNAs in peripheral blood mononuclear cells from subjects with LTBI and in healthy control individuals. Functional annotation prediction of miRNA target genes and pathway enrichment analyses were used to explore the putative links between these miRNAs and LTBI.

ORGANISM(S): Homo sapiens

SUBMITTER: Chun Zhang

PROVIDER: E-GEOD-54630 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Identification of latent tuberculosis infection-related microRNAs in human U937 macrophages expressing Mycobacterium tuberculosis Hsp16.3.

Meng Qing-Lin QL Liu Fei F Yang Xing-Yuan XY Liu Xiao-Mei XM Zhang Xia X Zhang Chun C Zhang Zong-De ZD

BMC microbiology 20140212

<h4>Background</h4>Latent tuberculosis infection (LTBI) relies on a homeostasis of macrophages and Mycobacterium tuberculosis (Mtb). The small heat shock protein, Mtb Hsp16.3 (also known as latency-associated antigen), plays an important role in Mtb persistence within macrophages. However, the mechanism of LTBI remains elusive. The aim of this study was to delineate LTBI-related miRNA expression in U937 macrophages expressing Mtb Hsp16.3 protein. U937 macrophages were infected with an integrase- ...[more]

PMID: 24521422

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Project description:From the results of microarray, 7616 miRNAs with significantly different expression ratios were selected when CNE-2-S-18/SP cells were treated with DC120. Among these, 80 miRNAs were up-regulated and 78 were down-regulated. We then chose 20 miRNAs through the qRT M-bM-^@M-^S PCR, and our results showed that 9 miRNAs were up-regulated after treatment with DC120, which was consistent with the results of the microarray. Among these, miR-30a were significantly up-regulated. These results indicated that miR 30a expression was up-regulated by DC120. We then investigated the role of miR 30a in DC120-treated cells, we explored its potential targets using a bioinformatics approach of complementary base pairing. The relative luciferase activity of the reporter that contained the wild type 3' UTR was significantly suppressed when miR 30a was contransfected. In contrast, the luciferase activity of the mutant reporter was unaffected by the contransfection of miR 30a, indicating that miR 30a suppressed Sox2 gene expression using the miR 30a binding sequence at the 3' UTR of the Sox2 gene. Additionally, the ectopic expression of miR 30a caused a decrease in Sox2 protein expression and mRNA levels in CNE-2-S-18 cells. These data demonstrate that miR 30a may directly target the Sox2 gene via seed matches on both 3' UTR. Taken together, our results demonstrate that DC120 regulates Sox2 via up-regulation of miR 30a. In this study, two samples were used to acquire the miRNA expression profiling. We analyzed the differences in miRNA profiles between control (DMSO solution) and DC120 treatment in CNE-2-S-18/SP cells from miRNA microarrays. The CNE-2-S-18/SP cells were separately cultured with a DMSO solution or 10M-NM-<mol/L DC120 for 15h, and total RNA from all of the cells was extracted. Then we chose 20 up-regulated miRNAs through the qRT M-bM-^@M-^S PCR, and our results showed that 9 miRNAs were up-regulated after treatment with DC120. Among these, miR-30a were significantly up-regulated. Then we investigated the role of miR 30a in DC120-treated cells by bioinformatic methods and Dual luciferase reporter system, the function of miR-30a was analyzed through ectopic expression of miR 30a. Taken together, our results demonstrate that DC120 regulates Sox2 via up-regulation of miR 30a.

Dataset Information

Identification of latent tuberculosis infection-related microRNAs in human U937 macrophages expressing Mycobacterium tuberculosis Hsp16.3

Publications

Identification of latent tuberculosis infection-related microRNAs in human U937 macrophages expressing Mycobacterium tuberculosis Hsp16.3.

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