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Small non-coding RNAs transfer through mammalian placenta and directly regulate fetal gene expression

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Protein & Cell 2015年第6期6卷 391-396页

作者： Jing Li Yujing Zhang Dameng Li Yuchen Liu Danping Chu Xiaohong Jiang Dongxia Hou ke zen Chen-Yu zhang State key Laboratory of Pharmaceutical Biotechnology Nanjing Advanced Institute for Life Sciences (NAILS) Nanjing University School of Life Sciences Jiangsu Engineering Research Center for MicroRNA Biology and Biotechnoiogy Nanjing University 22 Hankou Road Nanjing 210093 Jiangsu China

SMALL NON-CODING RNA CAN TRANSFER THROUGH THE PLACENTA AND REGULATE FETAL DEVELOPMENT Increasing evidence demonstrated that exogenous plant miRNAs in the diet can be absorbed and are found in mammalian tissues. These miRNAs influence the physio- logical and pathological progression of the recipient. In this issue, Jing Li and colleagues further reveal that exogenous/ endogenous small non-coding RNA in the maternal system can transfer through the placenta to the fetal side and influ- ence fetal development and health. These surprising finding demonstrate that maternal miRNAs profiling are very impor- tant to fetal development, which means maternal diet and the healthy status may influence fetal development and the postnatal morbidity via transplacental small non-coding RNAs. Furthermore, this finding also proposes a brand new potential strategy to treat fetal diseases in utero. Combining with gene therapy, maternal administrating represents a new trend for treating fetal diseases.

关键词： Amniotic Fluid Fetal Liver Fetal Development Maternal Plasma Rubella Virus

Small RNA existed in commercial reverse transcriptase： primary evidence of functional small RNAs

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Protein & Cell 2015年第1期6卷 1-5页

作者： Jie Xu Xi Chen Donghai Li Qun Chen Zhen Zhou Dongxia Hou Jin Wang Qipeng Zhang ke zen Chen-Yu Zhang Jiangsu Engineering Research Center for microRNA Biology and Biotechnology State Key Laboratory of Pharmaceutical Biotechnology School of Life Sciences Nanjing University Nanjing 210093 China

SMALL RNA IN REVERSE TRANSCRIPTASE： TOO SMALL TO BE FOUND Significant background in no-template or no-primer RT-PCR is frequently noted. However, the underlying mechanism is still unclear. In this issue, Jie Xu and colleagues reveal that the com- mercial reverse transcriptases contain small RNAs. In AMV reverse transcriptase, they detected microRNAs derived from the virus host chick or other birds. This surprising finding at least partially explains the high background in some AMV reverse transcriptases based RT-PCR.

关键词： Reverse Transcriptase Small RNAs miRNA Profile miRNA Expression Profile Solexa Sequencing

VlicroRNA-495 induces breast cancer ce migration by targeting JAM-A

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Protein & Cell 2014年第11期5卷 862-872页

作者： Minghui Cao Weiwei Nie Jing Li Yujing Zhang Xin Yan Xiaoxiang Guan Xi Chen ke zen Chen-yu Zhang Xiaohong Jiang Dongxia Hou State key Laboratory of Pharmaceutical Biotechnology Jiangsu Engineering Research Center for microRNA Biology and Biotechnology School of Life Sciences Nanjing University Nanjing 210093 China Department of Medical Oncology Jinling Hospital School of Medicine Southern Medical University Guangzhou 510282 China The Comprehensive Cancer Center of Drum Tower Hospital Medical School of Nanjing University and Clinical Cancer Institute of Nanjing University Nanjing 210008 China

MicroRNAs （miRNAs） are small, non-coding RNAs that function as post-transcriptional regulators of gene expression. The deregulated expression of miRNAs is associated with a variety of diseases, including breast cancer. In the present study, we found that miR-495 was markedly up-regulated in clinical breast cancer samples by quantitative real time-PCR （qRT-PCR）. Junctional adhesion molecule A （JAM-A） was predicted to be a potential target of miR-495 by bioinformatics analysis and was subsequently verified by luciferase assay and Western blotting. JAM-A was found to be negatively correlated with the migration of breast cancer cells through loss-of-function and gain-offunction assays, and the inhibition of JAM-A by miR- 495 promoted the migration of MCF-7 and MDA-MB-231 cells. Furthermore, overexpression of JAM-A could restore miR-495-induced breast cancer cell migration. Taken together, our findings suggest that miR-4g5 could facilitate breast cancer progression through the repression of JAM-A, making this miRNA a potential therapeutic target.

关键词： miR-495 JAM-A breast cancer migration

miR-10a inhibits cell proliferation and promotes cell apoptosis by targeting BCL6 in diffuse large B-cell lymphoma

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Protein & Cell 2016年第12期7卷 899-912页

作者： Qian Fan Xiangrui Meng Hongwei Liang Huilai Zhang Xianming Liu Lanfang Li Wei Li Wu Sun Haiyang Zhang ke zen Chen-Yu Zhang Zhen Zhou Xi Chen Yi Ba Department of Lymphoma Sino-US Center for Lymphoma and Leukemia Tianjin Medical University Cancer Institute and Hospital National Clinical Research Center of Cancer Key Laboratory of Cancer Prevention and Therapy Tianjin 300060 China State key Laboratory of Pharmaceutical Biotechnology NJU Advanced Institute of Life Sciences Jiangsu Engineering Research Center for MicroRNA Biology and Biotechnology School of Life Sciences Nanjing University Nanjing 210093 China Department of Digestion Tianjin Medical University Cancer Institute and Hospital National Clinical Research Center of Cancer Key Laboratory of Cancer Prevention and Therapy Tianjin 300060 China

The BCL6 （B-Cell Lymphoma 6） gene is a proto-onco- gene that is often expressed in diffuse large B-ceU lymphomas （DLBCLs）. BCL6 loss of function can kill DLBCL cells, demonstrating that BCL6 is necessary for the survival of DLBCL cells and could be a therapeutic target. In this study, we found that BCL6 protein levels were consistently upregulated in DLBCL tissues, whereas its mRNA levels varied randomly in tissues, suggesting that a post-transcriptional mechanism was involved in BCL6 regulation. We used bioinformatics analysis to search for miRNAs, which potentially target BCL6, and identified specific targeting sites for miR-10a in the 3＇-untranslated region （3＇-UTR） of BCL6. We fur- ther identified an inverse correlation between miR-10a levels and BCL6 protein levels, but not mRNA levels, in DLBCL tumor tissue samples. By overexpressing or knocking down miR-10a in DLBCL cells, we experi- mentally validated that miR-10a directly recognizes the 3＇-UTR of the BCL6 transcript and regulated BCL6expression. Furthermore, we demonstrated that nega- tively regulating BCL6 by miR-10a suppressed the pro- liferation and promoted apoptosis of DLBCL cells.

关键词： microRNA miR-10a BCL6 DLBCL proliferation apoptosis

Detection of the Pandemic H1N1/2009 Influenza A Virus by a Highly Sensitive Quantitative Real-time Reverse-transcription Polymerase Chain Reaction Assay

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Virologica Sinica 2013年第1期28卷 24-35页

作者： Zhu Yang Guoliang Mao Yujun Yuan-Chuan Chen Chengjing Liu Jun Luo Xihan Li ke zen Yanjun Pang Jianguo Wu Fenyong Liu Institute of Virology School of Life Sciences Nanjing University Taizhou Institute of Virology Jiangsu Affynigen Biotechnologies Inc. State key Laboratory of Virology College of Life Sciences Wuhan University College of Letters and Sciences University of Chicago Division of Infectious Diseases and Vaccinology School of Public Health University of California

A quantitative real time reverse-transcription polymerase chain reaction （qRT-PCR） assay with specific primers recommended by the World Health Organization （WHO） has been widely used successfully for detection and monitoring of the pandemic H1N1/2009 influenza A virus. In this study, we report the design and characterization of a novel set of primers to be used in a qRT-PCR assay for detecting the pandemic H1N1/2009 virus. The newly designed primers target three regions that are highly conserved among the hemagglutinin （HA） genes of the pandemic HlN1/2009 viruses and are different from those targeted by the WHO-recommended primers. The qRT-PCR assays with the newly designed primers are highly specific, and as specific as the WHO-recommended primers for detecting pandemic H1N1/2009 viruses and other influenza viruses including influenza B viruses and influenza A viruses of human, swine, and raccoon dog origin. Furthermore, the qRT-PCR assays with the newly designed primers appeared to be at least 10-fold more sensitive than those with the WHO-recommended primers as the detection limits of the assays with our primers and the WHO-recommended primers were 2.5 and 25 copies of target RNA per reaction, respectively. When tested with 83 clinical samples, 32 were detected to be positive using the qRT-PCR assays with our designed primers, while only 25 were positive by the assays with the WHO-recommended primers. These results suggest that the qRT-PCR system with the newly designed primers represent a highly sensitive assay for diagnosis of the pandemic HIN1/2009 virus infection.

关键词： Quantitative real time reverse-transcription polymerase chain reaction (qRT-PCR) Influenza A virus Detection

MiR-17 modulates proliferation and differentiation of neuroepithelial cells during mice corticogenesis

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MiR-17 modulates proliferation and differentiation of neuroe...

MicroRNA-23-27 clusters alleviate neuronal apoptosis though targeting Apaf-1 protein

中国神经科学学会第十届全国学术会议

作者： Su-Su MAO Han-Qin LI ke zen Chen-Yu ZHANG Liang LI Jiangsu Engineering Research Center for microRNA Biology and Biotechnology State Key Laboratory of Pharmaceutical BiotechnologyNanjing University School of Life Sciences Department of Virology University of California School of Public Health

MicroRNAs（miRNAs） are endogenously expressed small,non-coding nucleotides that repress gene expression at post-transcriptional ***,the developing brain contains a large diversity of miRNAs,indicating their crucial role in neural *** the present study,we analyzed miRNAs expression in mouse cortex at various developmental stages and found that miR-17 family was highly expressed in cortex at the early stages,which gradually decreased with the progress of *** investigation revealed that the change of miR-17-5p expression mainly occurred in ventricular zone/subventricular *** addition to its promotive effect on cell proliferation,miR-17-5p affects differentiation of neuroepithelial cells exposed to bone morphogenetic protein 2（BMP2）.Moreover,we demonstrated that such effect of miR-17-5p was mainly carried out through the modulation of BMPs signaling via repressing BMPR2 ***,these data suggest a pivotal role of miR-17 family in modulating cell activity in the early development of mouse cortex.

关键词： miR-17 neuroepithelial cell proliferation differentiation BMPR2

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MicroRNA-23-27 clusters alleviate neuronal apoptosis though ...

肾脏特异性microRNA的鉴定及功能预测

中国神经科学学会第十届全国学术会议

作者： Qun Chen Jie Xu Hanqin Li Liang Li Susu Mao ke zen Chen-Yu Zhang Qipeng Zhang Jiangsu Engineering Research Center for microRNA Biology and Biotechnology State Key Laboratory of Pharmaceutical BiotechnologySchool of Life SciencesNanjing University

Apoptotic protease activating factor-1（Apaf1） is a key apoptotic protein and plays a critical role in the regulation of neuronal *** of Apaf1 is gradually decreased during the central nervous system development,which responsible to the decreased caspase ***,the mechanism of decreased expression of Apaf1 in mouse cortex during development remains *** present study,we found that four microRNAs encoded by miR-23a-27a cluster and miR-23b-27b cluster participated in the regulation of Apaf1 *** assay and western blotting assay revealed that these four microRNAs suppressed the expression of Apaf1 in ***,the expression miR-23-27 clusters in mouse cortex are gradually increased,showed a reversed-correlated pattern to the Apaf1 expression *** injury,the most commonly cause of neurological damages in fetus during the perinatal period,could cause neuronal *** examined whether miR-23-27 cluster could attenuate the susceptibility of fetal *** primary neurons,miR-23-27 clusters inhibited the apoptosis induced by hypoxia in vitro;in neuronal specific transgenic mice,over-expression miR-23b-27b cluster inhibited the neuronal apoptosis induced by intrauterine hypoxia in *** together,our results indicate that miR-23-27 clusters are the endogenous inhibitory factors of Apaf1 expression;increased expression of miR-23-27 clusters may control the sensitivity of neurons to *** the first time,we discovered that microRNAs play roles in the neuronal apoptosis during *** study implied that microRNAs might be a potential target to treat the neuronal apoptosis diseases.

关键词： Apafl apoptosis neuron microRNA hypoxia

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肾脏病与透析肾移植杂志 2015年第2期24卷 107-112页

作者：孟娇李丽民刘志红曾科南京大学医学院附属金陵医院(南京军区南京总医院)肾脏科国家肾脏疾病临床医学研究中心全军肾脏病研究所硕士研究生南京210016 南京大学生命科学学院医药生物技术国家重点实验室江苏小核糖核酸工程研究中心

目的:应用Solexa技术分析和筛选出肾脏特异性表达microRNA(miRNA),同时利用生物信息学技术预测特异表达miRNA的靶基因并对其功能进行初步分析,探讨特异表达miRNA在肾脏中的功能。方法:选取8周龄C57/B6雄性小鼠,取其全身11个组织器官,提... 详细信息

目的:应用Solexa技术分析和筛选出肾脏特异性表达microRNA(miRNA),同时利用生物信息学技术预测特异表达miRNA的靶基因并对其功能进行初步分析,探讨特异表达miRNA在肾脏中的功能。方法:选取8周龄C57/B6雄性小鼠,取其全身11个组织器官,提取总RNA并从中分离小分子量RNA进行Solexa测序,筛选出相较其他组织器官,在肾脏中表达最为特异的miRNA;然后,利用实时荧光定量逆转录聚合酶链反应(qRTPCR)技术对筛选出的肾脏特异性表达miRNA进一步验证;最后,通过生物信息学技术对其进行靶基因预测,并对交集的基因集合分别进行GO分析和keGG通路分析。结果:Solexa测序结果显示在正常小鼠中miR-196a和miR-196b在肾脏最特异性高表达。qRT-PCR验证了miR-196a和miR-196b在小鼠11个组织器官中的表达分布情况,定量结果与测序结果相一致。TargetScan6.2和PicTar两个软件预测到交集靶基因74个。GO分析显示miR-196a/b的预测靶基因集合显著富集在血小板源生长因子结合活性、转录调控活性、细胞外基质的结构组成等分子功能上并参与调控代谢、发育等生物学过程(P<0.01)。信号转导通路显著富集于促性腺激素释放激素信号通路、细胞外基质受体相互作用等多个信号通路和脑胶质瘤、前列腺癌等疾病通路中(P<0.05)。结论:miR-196a/b在肾脏中特异性高表达,靶基因功能广泛,与细胞代谢、生长发育等密切相关。

关键词：肾组织Solexa测序 miR-196a miR-196b

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D-度量空间一组不动点定理

宝鸡文理学院学报（自然科学版） 2000年第2期20卷 98-100,149页

作者：尚增科宝鸡文理学院数学系陕西宝鸡721007

给出了 D

关键词： D-度量空间不动点自映射