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miRNA and Tm-3

Rebecca De Santis, Anke Liepelt, Jana C Mossanen, Anne Dueck, Nadine Simons, Antje Mohs, Christian Trautwein, Gunter Meister, Gernot Marx, Antje Ostareck-Lederer, Dirk H Ostareck
To secure the functionality of activated macrophages in the innate immune response, efficient life span control is required. Recognition of bacterial lipopolysaccharides (LPS) by toll-like receptor 4 (TLR4) induces downstream signaling pathways, which merge to induce the expression of cytokine genes and anti-apoptotic genes. MicroRNAs (miRNAs) have emerged as important inflammatory response modulators, but information about their functional impact on apoptosis is scarce. To identify miRNAs differentially expressed in response to LPS, cDNA libraries from untreated and LPS-activated murine macrophages were analyzed by deep sequencing and regulated miRNA expression was verified by Northern blotting and qPCR...
January 2, 2016: RNA Biology
Daoyang Zhou, Yingfeng Wan, Dajiang Xie, Yirong Wang, Junhua Wei, Qingfeng Yan, Peng Lu, Lianjie Mo, Jixi Xie, Shuxu Yang, Xuchen Qi
Although methyltransferase has been recognized as a major element that governs the epigenetic regulation of the genome during temozolomide (TMZ) chemotherapy in glioblastoma multiforme (GBM) patients, its regulatory effect on glioblastoma chemoresistance has not been well defined. This study investigated whether DNA methyltransferase (DNMT) expression was associated with TMZ sensitivity in glioma cells and elucidated the underlying mechanism. DNMT expression was analyzed by western blotting. miR-20a promoter methylation was evaluated by methylation-specific PCR...
2015: Experimental & Molecular Medicine
Zongyue Zeng, Ping Hu, Xi Tang, Hailong Zhang, Yane Du, Siyang Wen, Manran Liu
OBJECTIVE: To investigate the difference of miRNA expression levels of cancer-associated fibroblasts (CAFs) and normal fibroblasts (NFs) in human breast cancer microenvironment and its effect on the biological features of CAFs. METHODS: Collagenase-1 was used to digest the cancer and adjacent tissues to isolate CAFs and NFs. The isolated cells were cultured and characterized in purity and biological features. The expression of fibroblast secretory protein (FSP) in CAFs and NFs was detected by immunofluorescence staining and Western blotting...
October 2014: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
Qiong Xiang, Huifang Tang, Jia Yu, Jie Yin, Xiaoyan Yang, Xiaoyong Lei
This study was done to explore the role of microRNA-98 (miR-98) in cisplatin sensitization in human lung adenocarcinoma cell line. Differential expressions of miRNAs were analysed between cisplatin-resistant human lung adenocarcinoma cell line A549/DDP and its parental cell A549 by miRNAs microarray, of which 14 miRNAs were showed to be significantly (>2-fold) up-regulated and 8 miRNAs had marked down-regulation (<0.5-fold) in A549/DDP cells compared with in A549 cells. MiR-98, a member in the let-7 family, acts as a negative regulator in the expression of HMGA2 (high mobility group A2) oncogene, and it has been shown to have a nearly 3-fold decrease in A549/DDP cells...
April 2013: Die Pharmazie
Ling Wang, Bao-en Shan, Mei-xiang Sang, Yi-shui Lian, Bin Wang, Chun-yan Ding
OBJECTIVE: To explore the effect of microRNA (miRNA)-mediated p65 gene knockdown on the proliferation and apoptosis of human breast cancer MDA-MB-231 cells. METHODS: p65-targeted miRNA plasmid was constructed and transfected into MDA-MB-231 cells via lipofectamine(TM)2000. The expression of p65 gene in the transfected cells at the mRNA and protein levels were detected by RT-PCR and Western blotting, respectively. The cell proliferation and apoptosis were measured by MTT assay and flow cytometry (FCM), respectively...
October 2011: Nan Fang Yi Ke da Xue Xue Bao, Journal of Southern Medical University
Yevgeniya J Ioffe, Katherine B Chiappinelli, David G Mutch, Israel Zighelboim, Paul J Goodfellow
OBJECTIVES: The PTEN pseudogene, PTENP1, was recently shown to play a role in cell proliferation in a prostate cancer model. In the present study, we sought to determine whether PTENP1 is expressed in endometrial cancer (EMCA) cell lines and primary tumors along with the microRNAs (miRNAs) that are predicted to regulate PTEN and PTENP1 transcript levels. METHODS: RNA was prepared from six EMCA cell lines, three normal endometrial samples, and 61 primary tumors. TaqMan® RT-PCR was used to quantitate PTEN expression in all specimens and PTENP1 expression in cell lines, and normal endometrial (NE) samples...
February 2012: Gynecologic Oncology
Nick C T Schopman, Marcel Willemsen, Ying Poi Liu, Ted Bradley, Antoine van Kampen, Frank Baas, Ben Berkhout, Joost Haasnoot
Small virus-derived interfering RNAs (viRNAs) play an important role in antiviral defence in plants, insects and nematodes by triggering the RNA interference (RNAi) pathway. The role of RNAi as an antiviral defence mechanism in mammalian cells has been obscure due to the lack of viRNA detection. Although viRNAs from different mammalian viruses have recently been identified, their functions and possible impact on viral replication remain unknown. To identify viRNAs derived from HIV-1, we used the extremely sensitive SOLiD(TM) 3 Plus System to analyse viRNA accumulation in HIV-1-infected T lymphocytes...
January 2012: Nucleic Acids Research
Fang Huang, Xiao-yan Huang, Dong-sheng Yan, Xi Zhou, De-ye Yang
OBJECTIVE: To investigate the effects of microRNA-144 (miR-144) expression on H9C2 (2-1) myocytes. METHODS: MiR-144 was up-regulated in primary cultured H9C2 (2-1) myocytes through transfection. Cells transfected with Lipofectamine(TM) 2000 and its mixture with miRNA synthesized randomly as blank control and negative control respectively. The up-regulation of miR-144 was confirmed by real-time PCR. Cell apoptosis was evaluated by means of CCK-8, Caspase-3 and flow cytometry...
April 2011: Zhonghua Xin Xue Guan Bing za Zhi
W Shao, J-B Chen, F Wang, J-J Xia, Z-Q Qi
BACKGROUND: We sought to explore the effect of CD44 targeting on the tolerance to memory cell-mediated graft rejection. METHODS: We developed a cardiac transplantation model in nude mice and administered anti-CD44 monoclonal antibodies (mAbs) to these mice. Then, we used anti-CD44 mAb and CD44-interfering microRNA (miRNA) to inhibit CD44 expression in vitro. RESULTS: The median survival time (MST) associated with multiple intraperitoneal injections was >100 days, whereas that associated with CD4(+) Tm cells blocked CD44 and that associated with a single intraperitoneal injection of anti-CD44 mAb was 11 and 10...
September 2010: Transplantation Proceedings
D W Müller, A-K Bosserhoff
Although integrin beta(3) is known to play an important role in melanoma progression and invasion, regulation of integrin beta(3) expression in melanoma has not been analysed in detail until today. As transcriptional regulation of integrin beta(3) was ruled out by our analysis, we concentrated on the regulation by microRNAs (miRNAs). Comparing primary melanocytes and malignant melanoma cell lines, we found that one candidate miRNA, miR-let-7a, was lost in melanoma and sequence analysis suggested an interaction with the 3'-untranslated region (3'-UTR) of integrin beta(3) mRNA...
November 6, 2008: Oncogene
Mirco Castoldi, Sabine Schmidt, Vladimir Benes, Matthias W Hentze, Martina U Muckenthaler
MicroRNAs (miRNAs) represent a class of short (22 nt) noncoding RNAs that control gene expression post-transcriptionally. Microarray technology is frequently applied to monitor miRNA expression levels but is challenged by (i) the short length of miRNAs that offers little sequence for appending detection molecules; (ii) low copy number of some miRNA; and (iii) a wide range of predicted melting temperatures (Tm) versus their DNA complementary sequences. We recently developed a microarray platform for genome-wide profiling of miRNAs (miChip) by applying locked nucleic acid (LNA)-modified capture probes...
2008: Nature Protocols
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