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Elevated miR-155 expression induces immunosuppression via CD39⁺ regulatory T-cells in sepsis patient

INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES 2015年 40卷 135-141页

作者： Liu, Jingquan Shi, Kai Chen, Minhua Xu, Liang Hong, Jun Hu, Bangchuan Yang, Xianghong Sun, Renhua Zhejiang Prov Peoples Hosp Dept Intens Care Unit Hangzhou 310014 Zhejiang Peoples R China Hangzhou Normal Univ Affiliated Hosp Dept Resp Med Peoples Hosp Hangzhou 2 Hangzhou Zhejiang Peoples R China

Background: An altered microrna profile exists in many infectious diseases, including sepsis. CD39(+) regulatory T-cells (Tregs) have a remarkable immunosuppressive effect and play an important role in the regulation of immune balance in sepsis. However, the correlation between microrna changes and the ratio of CD39(+) Tregs in sepsis patients has not yet been reported. The altered microrna expression profile in sepsis patients was analyzed in this study. Moreover, the correlation between micrornas and disease severity and prognosis was investigated. Furthermore, the correlation between micrornas and the percentage of peripheral blood CD39(+) Tregs was investigated and further verified in an animal model. Methods: Sixty sepsis patients and 30 healthy controls were included. The difference in microrna expression was investigated by microrna microarray and was further confirmed by real-time quantitative PCR. The correlations between microrna changes and the Sepsis-related Organ Failure Assessment (SOFA) score, severity of sepsis, and survival were analyzed. The percentage CD39(+) Tregs in the peripheral blood of sepsis patients was measured by flow cytometry. The correlation between micrornas and the percentage CD39(+) Tregs was analyzed and further confirmed in a mouse sepsis model. Results: Compared to healthy controls, sepsis patients exhibited a significantly elevated microrna-155 (miR-155) level (p < 0.05), which was positively related to a higher SOFA score (r = 0.641, p < 0.05) and a greater severity of sepsis. The area under the receiver operating characteristic curve of miR-155 used for the prediction of 28-day survival was 0.763, with a cut-off point of 2.47. Patients with a miR-155 level >2.47 had a lower 28-day survival (p < 0.05). The miR-155 level of patients was proportional to the percentage of CD39(+) Tregs (r = 0.637, p < 0.05). After transfection with miR-155 inhibitor, the ratio of CD39(+) Tregs in mice with sepsis was significantly reduced (p < 0.

关键词： Sepsis microrna Regulatory T-cell microrna microarray Outcome

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Dysregulated microrna expression in adenocarcinoma of the uterine cervix: Clinical impact of miR-363-3p

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GYNECOLOGIC ONCOLOGY 2014年第3期135卷 565-572页

作者： Park, Hyun Lee, Mi-jung Jeong, Ju-yeon Choi, Min Chul Jung, Sang Geun Joo, Won Duk Lee, Chan An, Hee Jung CHA Univ Dept Gynecol Oncol Songnam South Korea CHA Univ Inst Clin Res Songnam South Korea CHA Univ Coll Med Dept Pathol Songnam 463712 Gyeonggi Do South Korea

Objective. Adenocarcinoma (ACA) of the uterine cervix is increasing in incidence and currently accounts for approximately 20% of all cervical malignancies. micrornas (miRNAs) have been investigated as potential biomarkers of cervical cancer;however, their role in ACA remains unknown. Here, we characterized miRNA expression profiles and investigated miRNAs as diagnostic and prognostic factors in ACA. Methods. Evaluation of genome-wide miRNA expression profiles in ACA by microarray led to the identification of ten candidate miRNAs, whose expression patterns were validated by qRT-PCR in 45 ACA, 10 normal control, and 15 squamous cell carcinoma samples. The association between miRNA expression and prognosis was analyzed in patients with ACA. Results. microarray analysis identified 86 miRNAs that were dysregulated more than 2.0-fold (p < 0.05) in ACA relative to normal tissues of the uterine cervix. Five most over- and underexpressed miRNAs were selected respectively and their expression patterns were confirmed in the validation set. MiR-135b, miR-192, and miR194 were overexpressed in ACA, and miR-363-3p, miR-195 and miR-199b were significantly associated with conventional prognostic factors. Overexpression of miR-363-3p by more than 2.5-fold relative to the normal control was a strong predictor of favorable prognosis (hazard ratio, 0.1;95% confidence interval, 0.009-0.779) after adjusting for confounders. Conclusions. MiR-135b, miR-192, and miR-194 are altered in uterine cervical ACA, and miR-363-3p is an independent favorable prognostic factor in ACA. These miRNAs could be of value as biomarkers for the diagnosis and prognosis of ACA. (C) 2014 Elsevier Inc. All rights reserved.

关键词： Cervical adenocarcinoma microrna microarray miR-363-3p Biomarker

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Elevated miR-155 expression induced immunosuppression via CD39+ Tregs in Sepsis patients

Elevated miR-155 expression induced immunosuppression via CD...

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中华医学会第二届重症心脏全国学术大会暨第三届西湖重症医学论坛、2015年浙江省重症医学学术年会

作者： Jingquan Liu Kai Shi Liang Xu Minhua Chen Xianghong Yang Jun Hong Bangchuan Hu Renhua Sun Department of Intensive Care Unit Zhejiang Provincial People's Hospital Department of Respiratory Medicine The Affiliated Hospital of Hangzhou Normal University(The 2nd People's Hospital of Hangzhou)

Background:Altered microrna profile exists in many infectious diseases such as *** a remarkable immunosuppressive effect,CD39 Tregs cells play an important role in the regulation of immune balance in *** the correlation between microrna change and the ratio of CD39 Tregs in sepsis patients has yet not been *** this study,the altered micrornas expression profile in sepsis patients was ***,the correlation between micrornas and severity of disease and prognosis was ***,the correlation between microrna and the percentage of peripheral blood CD39 Tregs was also investigated and further verified by animal ***:Sixty sepsis patients and 30 healthy controls were *** difference in microrna expression was investigated by microrna microarray and further confirmed by *** correlation between micrornas changes and SOFA score,severity of sepsis and survival was *** percentage of CD39+Tregs in peripheral blood of sepsis patients was measured by flow *** correlation between micrornas and percentage of CD39+Tregs was analyzed and further confirmed by mouse sepsis ***:Compared with the healthy controls,the sepsis patients exhibited a significant elevated microrna-155(miR-155)level(p<0.05),which related positively to higher SOFA score(r=0.641,p<0.05)and more severity of *** area under ROC curve of miR-155 used for prediction of 28 d survival was 0.763 with a cut-off point of *** with miR-155 level>2.47 had a lower 28 d survival(p<0.05).The miR-155 level of patients was proportional to the percentage of CD39+Tregs(r=0.637,p<0.05).After transfected with miR-155 inhibitor,the ratio of CD39+Tregs in mice with sepsis was significantly reduced(p<0.05).Conclusions:A higher level of miR-155 indicated a more severe condition and poor prognosis in sepsis *** possible underlying mechanism could be that miR-155 induced the percentage of CD39+Tregs and thus immunos

关键词： Sepsis microrna Regulatory T cell microrna microarray outcome

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Characterization of microrna expression profiles in 3T3-L1 adipocytes overexpressing C10orf116

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MOLECULAR BIOLOGY REPORTS 2013年第11期40卷 6469-6476页

作者： Qiu, Jie Zhou, Xiao-guang Zhou, Xiao-yu Zhu, Chun Shi, Chun-mei Ji, Chen-bo Cheng, Rui Li, Yong Guo, Xi-rong Nanjing Med Univ Nanjing Childrens Hosp Dept Newborn Infants Nanjing 210008 Peoples R China Nanjing Med Univ Nanjing Matern & Child Hlth Hosp Dept Pediat Nanjing 210004 Peoples R China Nanjing Med Univ Inst Pediat Nanjing 210029 Jiangsu Peoples R China

Our data in the previous report demonstrated that C10orf116 (AFRO) is an adipocyte lineage-specific nuclear factor that can modulate the master adipogenesis transcription factors early during differentiation. However, more precise functional properties of this gene need to be clarified and await further investigation. Therefore, in this study, we performed an expression profile of cellular micrornas (miRNAs) in the C10orf116 overexpression 3T3-L1 adipocytes and performed target prediction and functional enrichment of the differentially expressed miRNAs. Our study identified 34 miRNAs up-regulated in the 3T3-L1 adipocytes stably overexpressing C10orf116, whereas 43 miRNAs up-regulated in the control cells. The target genes of differentially expressed miRNAs were found to be involved in multiple signalling pathways, such as Wnt, TGF-beta, MAPK, Jak-STAT, insulin signalling pathway, et al. Our data provided novel information for the identification of C10orf116.

关键词： Obesity Adipocyte microrna microarray Target prediction Functional enrichment

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microrna detection by microarray

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ANALYTICAL AND BIOANALYTICAL CHEMISTRY 2009年第4期394卷 1117-1124页

作者： Li, Wei Ruan, Kangcheng Chinese Acad Sci Shanghai Inst Biol Sci Inst Biochem & Cell Biol State Key Lab Mol Biol Shanghai 200031 Peoples R China

micrornas (miRNAs) are a class of small noncoding RNAs similar to 22 nt in length that regulate gene expression and play fundamental roles in multiple biological processes, including cell differentiation, proliferation and apoptosis as well as disease processes. The study of miRNA has thus become a rapidly emerging field in life science. The detection of miRNA expression is a very important first step in miRNA exploration. Several methodologies, including cloning, northern blotting, real-time RT-PCR, microrna arrays and ISH (in situ hybridization), have been developed and applied successfully in miRNA profiling. This review discusses the main existing microrna detection technologies, while emphasizing microrna arrays.

关键词： microrna microrna detection microrna microarray microrna labeling

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microrna expression profiling of NGF-treated PC12 cells revealed a critical role for miR-221 in neuronal differentiation

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NEUROCHEMISTRY INTERNATIONAL 2012年第8期60卷 743-750页

作者： Hamada, Nanako Fujita, Yasunori Kojima, Toshio Kitamoto, Aya Akao, Yukihiro Nozawa, Yoshinori Ito, Masafumi Tokyo Metropolitan Inst Gerontol Res Team Mech Aging Itabashi Ku Tokyo 1730015 Japan Gifu Int Inst Biotechnol Dept Longev & Aging Res Gifu 5040838 Japan Hamamatsu Univ Sch Med Res Equipment Ctr Hamamatsu Shizuoka 4313192 Japan RIKEN Adv Sci Inst Computat Syst Biol Res Grp Yokohama Kanagawa 2300045 Japan Gifu Univ United Grad Sch Drug Discovery & Med Informat Sci Gifu 5011193 Japan Tokai Gakuin Univ Dept Food & Hlth Gifu 5048511 Japan

micrornas (miRNAs) are small non-coding RNAs that control protein expression through translational inhibition or mRNA degradation. MiRNAs have been implicated in diverse biological processes such as development, proliferation, apoptosis and differentiation. Upon treatment with nerve growth factor (NGF), rat pheochromocytoma PC12 cells elicit neurite outgrowth and differentiate into neuron-like cells. NGF plays a critical role not only in neuronal differentiation but also in protection against apoptosis. In an attempt to identify NGF-regulated miRNAs in PC12 cells, we performed miRNA microarray analysis using total RNA harvested from cells treated with NGF. In response to NGF treatment, expression of 8 and 12 miRNAs were up- and down-regulated, respectively. Quantitative RT-PCR analysis of 11 out of 20 miRNAs verified increased expression of miR-181a*, miR-221 and miR-326, and decreased expression of miR-106b*, miR-126, miR-139-3p, miR-143, miR-210 and miR-532-3p after NGF treatment, among which miR-221 was drastically up-regulated. Functional annotation analysis of potential target genes of 7 out of 9 miRNAs excluding the passenger strands (*) revealed that NGF may regulate expression of various genes by controlling miRNA expression, including those whose functions and processes are known to be related to NGF. Overexpression of miR-221 induced neuronal differentiation of PC12 cells in the absence of NGF treatment, and also enhanced neuronal differentiation caused by low-dose NGF. Furthermore, miR-221 potentiated formation of neurite network, which was associated with increased expression of synapsin I, a marker for synapse formation. More importantly, knockdown of miR-221 expression by antagomir attenuated NGF-mediated neuronal differentiation. Finally, miR-221 decreased expression of Foxo3a and Apaf-1, both of which are known to be involved in apoptosis in PC12 cells. Our results suggest that miR-221 plays a critical role in neuronal differentiation as well as prot

关键词： microrna microarray Nerve growth factor Neuronal differentiation PC12 cells miR-221

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Comparison of normalization methods with microrna microarray

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GENOMICS 2008年第2期92卷 122-128页

作者： Hua, You-Jia Tu, Kang Tang, Zhong-Yi Li, Yi-Xue Mao, Hua-Sheng Chinese Acad Sci Shanghai Inst Biol Sci Key Lab Syst Biol Ctr Funct GenomBioinformat Ctr Shanghai 200031 Peoples R China Natl Engn Ctr Biochip Shanghai 201203 Peoples R China Chinese Acad Sci Grad Sch Shanghai 200031 Peoples R China

micrornas (miRNAs) are a group of RNAs that play important roles in regulating gene expression and protein translation. In a previous study, we established an oligonucleotide microarray platform to detect miRNA expression. Because it contained only hundreds of probes, data normalization was difficult. In this study, the microarray data for eight miRNAs extracted from inflamed rat dorsal root ganglion (DRG) tissue were normalized using 15 methods and compared with the results of real-time polymerase chain reaction. It was found that the miRNA microarray data normalized by the print-tip loess method were the most consistent with results from real-time polymerase chain reaction. Moreover, the same pattern was also observed in 14 different types of rat tissue. This study compares a variety of normalization methods and will be helpful in the preprocessing of miRNA microarray data. Crown Copyright (c) 2008 Published by Elsevier Inc. All rights reserved.

关键词： microrna microarray normalization print-tip loess

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A global downregulation of micrornas occurs in human quiescent satellite cells during myogenesis

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DIFFERENTIATION 2012年第4期84卷 314-321页

作者： Koning, Merel Werker, Paul M. N. van Luyn, Marja J. A. Krenning, Guido Harmsen, Martin C. Univ Groningen Univ Med Ctr Groningen Dept Pathol & Med Biol NL-9711 GZ Groningen Netherlands Univ Groningen Univ Med Ctr Groningen Dept Plast Surg NL-9711 GZ Groningen Netherlands

During myogenesis, human satellite cells differentiate and form multinucleated myotubes, while a fraction of the human satellite cells enter quiescence. These quiescent satellite cells are able to activate, proliferate and contribute to muscle regeneration. Post-transcriptional regulation of myogenesis occurs through specific myogenic micrornas, also known as myomiRs. Although many micrornas are involved in myotube formation, little is known on the involvement of micrornas in satellite cells entering quiescence. This current study aims to investigate microrna involvement during differentiation of human satellite cells, specifically proliferating satellite cells entering quiescence. For this, clonally expanded human satellite cells were differentiated for 5 days, after which myotubes and quiescent satellite cells were separated through FACS sorting. Next, a microrna microarray comparison of proliferating satellite cells, myotubes and quiescent satellite cells was performed and verified through qRT-PCR. We show that during human satellite cell differentiation, micrornas are globally downregulated in quiescent satellite cells compared to proliferating satellite cells, in particular microrna-106b, microrna-25, microrna-29c and microrna-320c. Furthermore, we show that during myogenesis microrna-1, microrna-133, microrna-206 and microrna-486 are involved in myotube formation rather than satellite cells entering quiescence. Finally, we show an overall decrease in total mRNA in quiescent satellite cells, and an indication that RNaseL regulation plays a role in promoting and maintaining quiescence. Given the importance of quiescent satellite cells in skeletal muscle development and regenerative medicine, it is imperative to distinguish between myotubes and quiescent satellite cells when investigating skeletal muscle development, especially in microrna studies, since we show that micrornas are globally downregulated in quiescent human satellite cells. (c) 2012 International S

关键词： Satellite cells Skeletal muscle Quiescence microrna microrna microarray

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Simple and sensitive microrna labeling by terminal deoxynucleotidyl transferase

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生物化学与生物物理学报（英文版） 2012年第2期44卷 129-135页

作者： Botao Zhao Zhizheng Gong Zhongliang Ma Detao Wang Youxin Jin School of Life Sciences Shanghai University Shanghai 200444 China School of Life Sciences Shanghai University Shanghai 200444 China State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

micrornas (miRNAs) constitute a critically important class of non-translated, small RNAs, which post-transcriptionally regulate gene expression via one of the multiple mechanisms. To profile miRNA expression, microarrays have been extensively applied to the high-throughput detection of miRNAs. Here, we described a novel 3′-end miRNA-labeling method for microarray detection by terminal-deoxynucleotidyl transferase (TdT). TdT can catalyze the formation of polynucleotides at RNA receptor molecule with deoxycytidine triphosphate (dCTP). Using this activity, miRNA was successfully labeled by adding fluorescence dCTPs to its 3′-end. This labeling method was very simple and sensitive. The TdT-labeling method can detect as little as 0.04 fmol of synthetic small RNA, and produce precise and accurate measurements that span a linear dynamic range from 0.04 to 5 fmol of synthetic small RNA. The high consistency of miRNA expression data between our TdT method and real-time polymerase chain reaction analysis indicated the reliability and accuracy of the TdT method. Taken together, these results emphasize the immense potential application of the TdT-labeling method for sensitive and high-throughput microarray analysis of miRNA expression.

关键词： microrna microarray microrna labeling terminal deoxynucleotidyl transferase (TdT)

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Hybridization kinetics analysis of an oligonucleotide microarray for microrna detection

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生物化学与生物物理学报（英文版） 2011年第7期43卷 551-555页

作者： Botao Zhao Shuo Ding Wei Li Youxin Jin School of Life Sciences Shanghai University Shanghai 200444 China State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China School of Life Sciences Shanghai University Shanghai 200444 China State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

microrna (miRNA) microarrays have been successfully used for profiling miRNA expression in many physiological processes such as development, differentiation, oncogenesis, and other disease processes. Detecting miRNA by miRNA microarray is actually based on nucleic acid hybridization between target molecules and their corresponding complementary probes. Due to the small size and high degree of similarity among miRNA sequences, the hybridization condition must be carefully optimized to get specific and reliable signals. Previously, we reported a microarray platform to detect miRNA expression. In this study, we evaluated the sensitivity and specificity of our microarray platform. After systematic analysis, we determined an optimized hybridization condition with high sensitivity and specificity for miRNA detection. Our results would be helpful for other hybridization-based miRNA detection methods, such as northern blot and nuclease protection assay.

关键词： microrna microarray microrna

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