hsa-miR-100的生物信息学分析

目的 对hsa-miR-100进行靶基因、功能预测等生物信息学分析,为深入研究hsa-miR-100功能提供理论和试验基础.方法 利用Pubmed检索miRNA-100相关文章,通过miRBase、NCBI、UCSC Browser等在线工具分析hsa-miR-100序列,应用miRanda、TargetScan及PicTar预测hsa-miR-100靶基因,结合已证实的靶基因进行功能富集分析和信号通路富集分析.结果 hsa-miR-100与多种肿瘤发生、发展有关,其序列在各物种间具有高度保守性.hsa-miR-100靶基因功能富集于基因沉默、染色质沉默、细胞生物合成负性调节等(P<0.01),涉及肿瘤信号通路、溶酶体信号通路、凋亡信号通路等信号转导通路(P<0.05).结论 hsa-miR-100可能参与肿瘤发生相关的生物学过程.     

芪莲化积合剂对肝癌H22荷瘤小鼠抑瘤作用及miRNA122表达的影响

目的 观察芪莲化积合剂对肝癌H22荷瘤小鼠的抗肿瘤作用.方法 建立肝癌H22荷瘤小鼠实体瘤模型,将24只小鼠分为3组,对照组(生理盐水组)、芪莲化积合剂组和5-氟脲嘧啶(5-Fu)组各8只.通过测定荷瘤小鼠的瘤重、抑瘤率观察芪莲化积合剂剂对肿瘤生长的影响,采用RT-PCR技术检测miRNA122的表达情况.结果 芪莲化...     

乳腺癌患者外周血CD4~+CD25~+与CD8~+CD122~+调节性T细胞水平变化及临床意义

目的 探讨乳腺癌患者外周血中CD4+ CD25+ 与CD8+ CD122+ 调节性T细胞(Tregs)的分布情况及临床意义.方法 采用流式细胞仪测定65例乳腺癌患者外周血中的CD4+ CD25+ 与CD8+ CD122+ 调节性T细胞水平,结果与18例正常健康人(正常对照组)对比.结果 乳腺癌患者外周血中的CD4+CD25+与CD8+CD122+调节性T细胞均显著高于对照组(P<0.01);且与乳腺癌的分期呈正相关.结论 乳腺癌患者外周血中具有免疫负调控功能的CD4+ CD25+ 与CD8+ CD122+ 调节性T细胞水平较高,可能与乳腺癌的病情及预后相关.     

New therapeutic opportunities for hepatitis C based on small RNA

Hepatitis C virus (HCV) infection is one of the major causes of chronic liver disease, including cirrhosis and liver cancer and is therefore, the most common indication for liver transplantation. Conventional antiviral drugs such as pegylated interferon-alpha, taken in combination with ribavirin, represent a milestone in the therapy of this disease. However, due to different viral and host factors, clinical success can be achieved only in approximately half of patients, making urgent the requirement of exploiting alternative approaches for HCV therapy. Fortunately, recent advances in the understanding of HCV viral replication and host cell interactions have opened new possibilities for therapeutic intervention. The most recent technologies, such as small interference RNA mediated gene-silencing, anti-sense oligonucleotides (ASO), or viral vector based gene delivery systems, have paved the way to develop novel therapeutic modalities for HCV. In this review, we outline the application of these technologies in the context of HCV therapy. In particular, we will focus on the newly defined role of cellular microRNA (miR-122) in viral replication and discuss its potential for HCV molecular therapy.     

Efficient inhibition of human cytomegalovirus UL122 gene expression in cell by small interfering RNAs

In order to develop a gene therapy to human cytomegalovirus (HCMV), RNA interference (RNAi) was employed to inhibit the expression of HCMV UL122 gene in vitro. Recombinant vector pUL122‐EGFP, which expressed UL122‐EGFP fusion protein, and recombinant vectors psi122‐1, psi122‐2 and psi122‐3, which expressed small interfering RNAs (siRNAs) targeted to UL122 were contransfected into AD293 cells. The fluorescence signal of pUL122‐EGFP was greatly suppressed by psi122‐1 and psi122‐2, with an inhibitory rate of 82.0% ± 1.0% and 79.5% ± 2.5%, respectively. The mRNA of pUL122‐EGFP of the cells transfected with psi122‐1 and psi122‐2 was decreased 97.3% ± 0.6% and 98.0% ± 0.1%, respectively. Vector psi122‐3 showed a slightly low suppression rate. Therefore, it may be concluded that plasmids encoding siRNAs targeted to UL122 is able to in vitro reduce markedly the expression of UL122‐EGFP. And it is very likely that the psi122‐1 and psi122‐2 are potentially efficacious siRNAs in the gene therapy of HCMV infection in vivo, in which further investigations are required. This study is expected to greatly facilitate the use of the RNAi technology for the anti‐HCMV studies. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

用聚乙二醇分离富集microRNA的方法探讨

目的 探讨用聚乙二醇(polyethylene glycol,PEG)溶液分离富集miRNA的操作方法和分离富集效果,并与Ambion公司的miRNA分离试剂盒分离效果进行比较.方法 用PEG溶液和Ambion公司的miRNA分离试剂盒从肝脏组织总RNA中分离富集miRNA,用变性琼脂糖和变性聚丙烯酰胺凝胶电泳鉴定分离效果,并在富集的miRNA中用RT-PCR扩增miR-122以鉴定是否有效地回收了miRNA.结果 PEG和Ambion公司的miRNA分离试剂盒都能有效地富集miRNA,PEG富集的RNA片段比Ambion公司的试剂盒的大.结论 PEG溶液能有效地分离富集miRNA,和Ambion公司的miRNA分离试剂盒分离效果相当,并具有操作简便、快捷及成本低廉的优点.     

miRNA-122a靶基因预测及生物信息学分析

目的:利用基因芯片技术分析肝癌HepG2细胞和正常肝上皮LO2细胞中miRNA的表达,并对HepG2细胞中低表达的miRNA-122a进行靶基因预测及相关生物信息学分析,为以miRNA-122a为靶点的基因治疗提供理论和实验基础.方法:利用基因芯片技术检测HepG2细胞和LO2细胞中miRNA-122a表达水平,通过生物信息学预测miRNA-122a的靶基因,并对其靶基因进行功能富集分析(GO-analysis)、信号转导通路富集分析(Pathway-analysis)和蛋白质相互作用网络分析.结果:与LO2细胞比较,miRNA-122a在HepG2细胞中呈低表达.miRNA-122a预测靶基因有1 104个,其靶基因集合功能分别富集于碳水化合物生物合成、核苷酸代谢、细胞因子受体结合、细胞周期等生物学过程(P<0.001);信号转导通路显著富集于JAK-STAT信号通路、Wnt信号通路、MAPK信号通路、ErbB信号通路、细胞周期等信号转导通路(P<0.001).结论:miRNA-122a在HepG2细胞中呈现低表达,miRNA-122a预测靶基因集合显著富集在与肿瘤发生相关的信号通路中.     

Prevalence of the amyloidogenic transthyretin (TTR) V122I allele in 14 333 African–Americans

Background: Transthyretin (TTR) V122I (rs76992529) is one of 111 variants caused by point mutations in the coding sequence of the human TTR gene that are associated with systemic amyloidosis. It results from a G to A transition at a CG dinucleotide in codon 142(122 of the mature protein) of the gene and has been described almost exclusively in people of African descent. Several series have reported allele frequencies from 0.015 to 0.020 in African-Americans.

Objective: To define more accurately the frequency of the TTR V122I variant allele in the African-American population.

Methods: DNA isolated from blood spots from 1688 New York State African-American newborns was genotyped for the TTR V122I allele. We also compiled new data from the Jackson Heart Study and previously unpublished data from the Dallas Heart Study, plus data from a San Diego “wellness study”, providing 15?650 additional allelotypes to those already reported.

Results: Among the New York newborns, the TTR V122I allele was present in 65/3376 alleles (allele prevalence 0.0193). The combined available data from all the non-selected African-American cohorts showed the TTR variant allele to be present in 451/26?062 alleles (allele prevalence of 0.0173), slightly but not significantly lower than our previously published estimates.

Conclusions: The allele prevalence for TTR V122I in African-Americans is 0.0173. Of African–Americans under age 65, 3.43% carry at least one copy of the variant amyloidogenic allele.