In vivo discovery of a peptide that prevents CUG-RNA hairpin formation and reverses RNA toxicity in myotonic dystrophy models.
摘 要： 目的探讨非小细胞型肺癌组织中CUG结合蛋白1（CUGBP1）mRNA及蛋白的表达以及与预后的关系。方法选取2009年7月至2011年4月在青岛大学附属医院胸外科接受手术治疗的57例非小细胞型肺癌患者，男32例，女25例；年龄43～74（60.6±8.9）岁。通过半定量逆转录一聚合酶链反应（RT-PCR）及免疫组织化学技术检测癌组织及周围正常组织CUGBP1的基因和蛋白表达情况。所有患者术后每2个月电话随访1次，采用肿瘤进展时间（TTP）作为观察评估指标，分析CUGBP1 mRNA及临床病理各指标与预后的关系。采用χ^2检验对CUGBP1 mRNA及蛋白的表达进行统计分析，采用COX单因素及多因素分析CUGBP1 mRNA对于肺癌预后的意义。结果CUGBP1 mRNA及CUGBP1蛋白在肺癌组织中高表达，并且与TNM分期及分化密切相关。通过单因素及多因素分析，CUGBP1 mRNA[P=0.0074，HR=3.701，95％CI（1.420，9.648）]、TNM分期[P〈0.0001，HR=4.043，95％CI（2.098，7.794）]及年龄（P=0.0018，HR=3.207，95％CI（1.544，6.664）]可以作为非小细胞肺癌患者的独立预后因素。结论CUGBP1 mRNA及蛋白在肺癌组织中高表达。作为非小细胞肺癌患者的独立预后因素，CUGBP1 mRNA高表达患者预后较差。
Objective To detect mRNA and protein expression of CUG-binding protein 1(CUGBP1)in non-small cell lung cancer(NSCLC)，and assess the prognostic signifi cance of CUGBP1. Methods Fifty-seven NSCLC patients who received surgical resection at the Department of Thoracic Surgery of Affiliated Hospital of Qingdao University between July 2009 and April 2011 were enrolled in this study. There were 32 male and 25 female patients with their age of 43-74(60.6±8.9) years. The expressions of CUGBP1 mRNA and protein in tumor and adjacent normal tissues were detected by semi-quantitative RT-PCR and immunohistochemistry. All the patients were followed up every 2 months after discharge via the phone. Time to progression(TTP) was used to evaluate the relationship between CUGBP1 mRNA，clinicopathological variables and prognosis. The percentage of CUGBP1 mRNA and CUGBP1 expression was correlated with clinical characteristics using χ2 test. The prognostic signifi cance of CUGBP1 mRNA was assessed by univariate and multivariate analysis using Cox regression analysis. Results The expressions of CUGBP1 mRNA and CUGBP1 were over-expressed in cancer tissue， and were signifi cantly correlated with TNM-stage and differentiation. Univariate and multivariate analysis showed that CUGBP1 mRNA expression(P=0.0074，HR=3.701，95% CI 1.420-9.648)，TNM-stage(P 0.000 1，HR= 4.043，95% CI 2.098-7.794) and age(P=0.0018，HR=3.207，95% CI 1.544-6.664) were independent predictors of postsurgical survival in NSCLC patients. Conclusions CUGBP1 mRNA and CUGBP1 are over-expressed in NSCLC，and over-expression of CUGBP1 mRNA independently predicts a shorter postsurgical survival in NSCLC patients.
Recent evidence suggests that a certain type of () is hierarchically organized by a subset of cells with stem cell features (stem cells; ). Although normal stem cells and are considered to share similar self-renewal programs， it remains unclear whether differentiation programs are also maintained in and effectively used for eradication. In this study， we investigated the effect of oncostatin M ()， an -related cytokine known to induce the differentiation of hepatoblasts into hepatocytes， on liver . receptor expression was detected in the majority of epithelial molecule-positive (EpCAM(+)) with stem/progenitor cell features. treatment resulted in the induction of hepatocytic differentiation of EpCAM(+) cells by inducing signal transducer and activator of 3 activation， as determined by a decrease in stemness-related gene expression， a decrease in EpCAM， alpha-fetoprotein and protein expressions， and an increase in albumin protein expression. -treated EpCAM(+) cells showed enhanced with expansion of the EpCAM-negative non-population. Noticeably， combination of treatment with the chemotherapeutic agent (5-FU)， which eradicates EpCAM-negative non-， dramatically increased the number of apoptotic cells in vitro and suppressed in vivo compared with either saline control， ， or 5-FU treatment alone. Taken together， our data suggest that could be effectively used for the differentiation and active of dormant EpCAM(+) liver ， and the combination of and conventional chemotherapy with 5-FU efficiently eliminates by targeting both and non-.
摘 要： 目的：探讨CUG连接蛋白1（CUG-Binding Protein 1，CUGBP1）基因在前列腺癌的表达及其蛋白功能。方法：用定量RT-PCR法在前列腺癌组织和癌旁组织中检测CUGBP的mRNA水平。Western blot法检测4种前列腺癌细胞株（22RV1，PC-3，DU145，LNCaP clone FGC）中CUGBP1的表达。利用RNA干扰技术检测PC-3细胞在CUGBP1基因沉默后细胞的增殖与凋亡状态。利用定量RT-PCR法确定其下游调控基因的mRNA水平。结果：CUGBP1在PC-3和22RV1细胞株中高表达。CUGBP1的mRNA在前列腺癌组织中较癌旁组织显著增高。CUGBP1基因shRNA导致CUGBP1基因沉默后抑制了PC-3细胞株的增殖并诱导其凋亡;同时，使前列腺癌细胞在细胞周期上发生G1期阻滞。CUGBP1基因被干扰以后，PC-3细胞株中CDKN1A的mRNA水平发生明显上调，同时BCL2、PCNA、MCM2和MCM3基因的mRNA水平发生明显下调。结论：CUGBP1基因和前列腺癌相关。CUGBP1基因可能通过上调/下调下游基因来调节前列腺癌细胞的凋亡，改变细胞的增殖和细胞周期。
Abstract Myotonic dystrophy is the most common form of adult-onset muscular dystrophy， originating in a CTG repeat expansion in the DMPK gene. The expanded CUG transcript sequesters MBNL1， a key regulator of alternative splicing， leading to the misregulation of numerous pre-mRNAs. We report an RNA-targeted agent as a possible lead compound for the treatment of myotonic dystrophy type 1 (DM1) that reveals both the promise and challenges for this type of small-molecule approach. The agent is a potent inhibitor of the MBNL1–rCUG complex with an inhibition constant ( K i ) of 25±8 n m ， and is also relatively nontoxic to HeLa cells， able to dissolve nuclear foci， and correct the insulin receptor splicing defect in DM1 model cells. Moreover， treatment with this compound improves two separate disease phenotypes in a Drosophila model of DM1： adult external eye degeneration and larval crawling defect. However， the compound has a relatively low maximum tolerated dose in mice， and its cell uptake may be limited， providing insight into directions for future development.
Abstract Background ： CUG-BP， Elav-like family member 1 (CELF1) is a multifunctional RNA binding protein found in a variety of adult and embryonic tissues. In the heart， CELF1 is found exclusively in the myocardium. However， the roles of CELF1 during cardiac development have not been completely elucidated. Results ： Myofibrillar organization is disrupted and proliferation is reduced following knockdown of CELF1 in cultured chicken primary embryonic cardiomyocytes. In vivo knockdown of Celf1 in developing Xenopus laevis embryos resulted in myofibrillar disorganization and a trend towards reduced proliferation in heart muscle， indicating conserved roles for CELF1 orthologs in embryonic cardiomyocytes. Loss of Celf1 also resulted in morphogenetic abnormalities in the developing heart and gut. Using optical coherence tomography， we showed that cardiac contraction was impaired following depletion of Celf1， while heart rhythm remained unperturbed. In contrast to cardiac muscle， loss of Celf1 did not disrupt myofibril organization in skeletal muscle cells， although it did lead to fragmentation of skeletal muscle bundles. Conclusions ： CELF1 is required for normal myofibril organization， proliferation， morphogenesis， and contractile performance in the developing myocardium. This article is protected by copyright. All rights reserved.
摘 要： 目的：探讨CUG连接蛋白1（CUG-binding protein 1，CUGBP1）在膀胱癌组织中的表达及其表达对膀胱癌BIU-87细胞株增殖及迁移能力的影响。方法：采用免疫组化及western blot检测65例膀胱癌及相应的癌旁正常组织中CUGBP1的表达，并探讨CUGBP1表达与膀胱癌患者临床病理特征的关系。同时，采用siRNA特性干扰片段下调CUGBP1在膀胱癌细胞株BIU-87中的表达.探讨CUGBP1下调对细胞增殖及侵袭能力的影响。结果：免疫组化结果显示CUGBP1在膀胱癌组织中的阳性表达率为58.5％，高于癌旁正常组织（9.2％），差异具有统计学意义（P〈0.01）；Western blot结果也显示CUGBP1在膀胱癌组织中的表达高于癌旁正常组织。同时。CUGBP1在膀胱癌组织中的表达与淋巴结转移、TNM分期及分化程度有关（P〈0.05）。此外，BIU-87细胞株中CUGBP1表达明显下调后，细胞的增殖及侵袭能力均明显下降（P〈0.05）。结论：CUGBP1表达与膀胱癌的形成相关。其表达下调可明显抑制细胞增殖及侵袭。
Abstract OBJECTIVE To identify linear determinants of human aquaporin 4 (hAQP4) in the context of HLA-DRB1*03：01. DESIGN In this controlled study with humanized experimental animals， HLA-DRB1*03：01 transgenic mice were immunized with whole-protein hAQP4 emulsified in complete Freund adjuvant. To test T-cell responses， lymph node cells and splenocytes were cultured in vitro with synthetic peptides 20 amino acids long that overlap by 10 amino acids across the entirety of hAQP4. The frequency of interferon γ， interleukin (IL) 17， granulocyte-macrophage colony-stimulating factor， and IL-5-secreting CD4+ T cells was determined by the enzyme-linked immunosorbent sport assay. Quantitative immunofluorescence microscopy was performed to determine whether hAQP4281-300 inhibits the binding of anti-hAQP4 recombinant antibody to surface full-length hAQP4. SETTING Academic neuroimmunology laboratories. SUBJECTS Humanized HLA-DRB1*03：01+/+ H-2b-/- transgenic mice on a B10 background. RESULTS Peptide hAQP4281-300 generated a significantly (P <.01) greater TH1 and TH17 immune response than any of the other linear peptides screened. This 20mer peptide contains 2 dominant immunogenic 15mer peptides. hAQP4284-298 induced predominantly an IL-17 and granulocyte-macrophage colony-stimulating factor TH cell phenotype， whereas hAQP4285-299 resulted in a higher frequency of TH1 cells. hAQP4281-300 did not interfere with recombinant AQP4 autoantibody binding. CONCLUSIONS hAQP4281-330 is the dominant linear immunogenic determinant of hAQP4 in the context of HLA-DRB1*03：01. Within hAQP4281-330 are 2 dominant immunogenic determinants that induce differential TH phenotypes. hAQP4 determinants identified in this study can serve as diagnostic biomarkers in patients with neuromyelitis optica and may facilitate the monitoring of treatment responses to pharmacotherapies.
CUG2蛋白，又称着丝粒蛋白W，是一种近期发现的核蛋白，研究发现CUG2为细胞分裂、细胞凋亡等过程所必需，在许多癌组织中都存在高表达，如结肠直肠 癌及胃癌，被预测是一种致癌基因.它位于核仁上或者着丝粒上，能和着丝粒蛋白T相互结合，共同参与着丝粒染色质结构的形成.现概要介绍CUG2基因的发 现、结构特点、生物学功能及其与肿瘤之间关系的最新进展.