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国家自然科学基金(30700714)

作品数:3 被引量:41H指数:2
相关作者:王琳程勇前刘妍徐东平王春梅更多>>
相关机构:解放军第302医院中国人民解放军总医院北京地坛医院更多>>
发文基金:国家自然科学基金更多>>
相关领域:医药卫生生物学更多>>

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Altered expression profiles of microRNAs in a stable hepatitis B virus-expressing cell line被引量:38
2009年
Background MicroRNAs (miRNAs) are highly conserved small non-coding RNAs of 18-25 nucleotides (nt) that mediate post-transcriptional gene regulation. Hepatitis B virus (HBV) can cause either acute or chronic hepatitis B, and is a high risk factor for liver cirrhosis and hepatocellular carcinoma. Some mammalian viruses have been shown to modulate the expression of host cellular miRNAs. However, interactions between the HBV and the host cellular miRNAs are largely unknown.Methods miRNA microarray and Northern blotting analysis were used to compare the expression profile of cellular miRNAs of a stable HBV-expressing cell line HepG2.2.15 and its parent cell line HepG2. mRNA microarray assay and the miRanda program were used to predict the miRNA targets. A flow cytometric assay was further used to investigate the expression of human leukocyte antigen (HLA)-A.Results Eighteen miRNAs were differentially expressed between the two cell lines. Among them, eleven were up-regulated and seven were down-regulated in HepG2.2.15 cells. Northern blotting analysis confirmed that the expression of miR-181a, miR-181b, miR-200b and miR-146a were up-regulated and the expression of miR-15a was down-regulated, which was in consistent with the results of the microarray analysis. Furthermore, some putative miRNA targets were predicted and verified to be linked with mRNA expression. The 3′-UTR of HLA-A gene had one partially complementary site for miR-181a and miR-181 a might down-regulate the expression of HLA-A.Conclusion HBV replication modulates the expression of host cellular miRNAs, which may play a role in the pathogenesis of HBV-related liver diseases.
LIU YanZHAO Jian-JunWANG Chun-meiLI Mian-yangHAN PingWANG LinCHENG Yong-qianFabien ZoulimMA XuXU Dong-ping
关键词:MICROARRAY
miR-181a在转染乙型肝炎病毒基因组的HepG2.2.15细胞中的表达研究被引量:3
2008年
目的鉴定分析microRNA(miRNA)miR-181a在转染了乙型肝炎病毒(HBV)基因组的HepG2.2.15细胞中的表达情况,探讨miR-181a在与HBV相关的肝脏疾病中的作用。方法以本课题组基因芯片结果为基础,设计并合成miR-181a探针,采用Northernblotting检测miR-181a在HepG2.2.15和HepG2细胞(对照组)中的表达水平,应用生物信息学方法结合mRNA表达谱芯片结果预测miR-181a的靶基因,选取靶基因HLA-A2,应用流式细胞仪分析HLA-A2分子在上述2种细胞中的表达。结果Northernblotting结果显示,与对照组相比,miR-181a在HepG2.2.15细胞中的表达量显著增高;miR-181a可能的靶基因包括C8A、IDH1和HLA-A等,miR-181a可能通过其种子序列与其靶基因HLA-A的3′-UTR区部分互补结合来调节HLA-A的表达;流式分析也显示HLA-A2分子在HepG2.2.15细胞中的表达量(43.9%)显著低于HepG2细胞(96.6%)。结论miR-181a在HBV转染的HepG2.2.15细胞中高表达,并可能下调靶基因HLA-A的表达,推测这可能是HBV感染后病毒逃避免疫反应、持续复制的机制之一。
刘妍王春梅李绵洋王琳程勇前张玲霞徐东平
关键词:乙型肝炎病毒HLA抗原
钙离子调节亲环素配体及其不同剪接体的克隆及生物信息学分析
2008年
目的:克隆钙离子调节亲环素配体(CAML)及其不同剪接体,并应用生物信息学进行分析。方法:以Jurkat、HepG2细胞提取的cDNA为模板,PCR扩增CAML基因,选用pGEM-T-easy载体进行TA克隆,利用VectorNTI9.0软件与CAML基因所在染色体进行比对分析,并对CAML全长基因进行生物信息学分析。结果:CAML基因克隆成功,并证实CAML基因在细胞中存在3种剪接体,分别为缺失全长CAML cDNA序列中634~698、173~633和173~698 bp的CAML序列,大小分别为825、429和363 bp。这些剪接体的组成中均包含了CAML与其他蛋白相互结合的亲水的N末端,以及CAML发挥调节钙离子内流作用所必需的第2、3跨膜结构域,说明这些剪接体均具有发挥CAML基本功能的可能。结论:成功克隆了CAML基因,并首次证实CAML基因在细胞中存在不同剪接体。
程勇前成军刘妍王琳徐东平钟彦伟曲建慧赵平
关键词:剪接体生物信息学分析
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