MicroRNA signatures characterize diffuse large B-cell lymphomas and follicular lymphomas

MicroRNAs (miRNA, miR) are negative regulators of gene expression that play an important role in diverse biological processes such as development, cell growth, apoptosis and haematopoiesis, suggesting their association with cancer. Here we analysed the expression signatures of 157 miRNAs in 58 diffuse large B-cell lymphoma (DLBCL), 46 follicular lymphoma (FL) and seven non-neoplastic lymph nodes (LN). Comparison of the possible combinations of DLBCL-, FL- and LN resulted in specific DLBCL- and FL-signatures, which include miRNAs with previously published function in haematopoiesis ( MIRN150 and MIRN155 ) or tumour development ( MIRN210 , MIRN10A , MIRN17-5P and MIRN145 ). As compared to LN, some miRNAs are differentially regulated in both lymphoma types ( MIRN155 , MIRN210 , MIRN106A , MIRN149 and MIRN139 ). Conversely, some miRNAs show lymphoma-specific aberrant expression, such as MIRN9/9* , MIRN301 , MIRN338 and MIRN213 in FL and MIRN150 , MIRN17-5P , MIRN145 , MIRN328 and others in DLBCL. A classification tree was computed using four miRNAs ( MIRN330 , MIRN17-5P , MIRN106a and MIRN210 ) to correctly identify 98% of all 111 cases that were analysed in this study. Finally, eight miRNAs were found to correlate with event-free and overall survival in DLBCL including known tumour suppressors ( MIRN21 , MIRN127 and MIRN34a ) and oncogenes ( MIRN195 and MIRNLET7G ).     

子痫前期患者血清微小RNAs的差异表达研究

目的探讨子痫前期患者血清微小RNAs(miRNAs)不同的表达水平,分析子痫前期相关标志物。方法选择10个子痫前期患者胎盘中差异表达的miRNA,分别收集该院30例子痫前期患者和30例正常妊娠孕妇孕早期(12~14周)、孕中期(20~24周)和孕晚期(28~32周)血清miRNAs,采用SYBR Green qRT-PCR法检测血清中10个miRNAs的表达量。结果子痫前期胎盘中高表达的3个miRNAs(miR-152、miR-183、miR-210)在子痫前期孕中、晚期血清中表达显著升高,而胎盘中高表达的miR-182仅在子痫前期孕晚期表达显著升高;子痫前期胎盘中低表达的6个miRNAs(miR-1、miR-328、miR-363、miR-377、miR-500、miR-584)在子痫前期各个孕期的表达,差异均无统计学意义(P0.05)。结论 miR-152、miR-183、miR-210在子痫前期孕中、晚期血清中表达显著升高,可能是预测子痫前期发生的良好生物标志物。     

miRNA在前列腺癌中的研究进展

前列腺癌(PCa)是男性泌尿系最常见的恶性肿瘤之一,微小RNA(miRNA)是一类内源性的非编码小RNA,研究发现miRNA与PCa的发生和发展密切相关,多种miRNA在PCa中表达异常。本文通过描述miRNA在PCa中表达差异及其与预后的相关性,进一步分析miRNA与放化疗、雄激素受体,以及PCa转移的相关性,来阐明miRNA在PCa发生发展中的的作用。     

microRNA-1271靶向调控白血病细胞CYLD蛋白的表达

目的探讨人白血病细胞中微小RNA(microRNA)-1271对CYLD蛋白表达的调控作用。方法 qRT-PCR检测microRNA-1271在不同的人白血病细胞系、临床初诊未治的几种类型原代白血病细胞、正常人外周血单个核细胞中的表达差异,利用 Targetscan 信息学预测软件预测 microRNA-1271靶向CYLD基因,构建携带靶基因野生型及突变型3’非翻译区(3’UTR)(缺失了整段预测的microRNA-1271结合序列)的双荧光素酶报告基因质粒,采用脂质体 Lipo-fectamine 3000包裹双荧光素酶重组质粒及 microRNA-1271模拟物(mimic)或阴性对照,共转染HEK293A细胞,应用双荧光素酶报告基因检测试剂盒测定荧光素酶活性。 FAM标记的microRNA-1271抑制物和阴性对照分别核转染至人白血病细胞系K562细胞,Western blot法检测CYLD蛋白水平的表达变化。结果 microRNA-1271在不同人白血病细胞系以及临床初诊未治的原代白血病细胞中的表达均明显高于正常人外周血单个核细胞,双荧光素酶报告基因实验验证CYLD是 microRNA-1271的潜在靶基因;K562细胞中转染microRNA-1271抑制物下调 microRNA-1271后, Western blot法检测结果显示CYLD蛋白水平明显上调。结论人白血病细胞中microRNA-1271的表达上调,下调microRNA-1271后可以促进靶基因CYLD蛋白的表达, microRNA-1271有可能成为白血病治疗的新靶点。     

MicroRNA基因递送系统抗激素非依赖型前列腺癌增殖作用研究

目的：采用分枝状聚乙烯亚胺（BPEI）、线型的聚乙烯亚胺（LPEI）和树枝状聚合物聚酰胺-胺（PAMAM）与具有抑制激素非依赖型前列腺癌细胞PC3增殖的miRNA-15a和miRNA-16-1质粒结合,构建了MicroRNA基因递送系统。考察形成的三种纳米复合物的粒径、zeta电位、细胞内摄取,以及对PC3细胞的抑制作用。方法：利用粒径分析仪测定形成的三种纳米复合物的粒径和电位,凝胶电泳实验测定三种材料对miRNA的结合能力; 利用FAM标记的NC-miRNA考察PC3细胞对三种复合的摄取情况;通过CCK8 法测定三种材料与miRNA-15a和miRNA-16-1形成的复合物对PC3细胞的抑制作用;PCR法测定三种材料与miRNA-15a和miRNA-16-1结合后对PC3细胞中Bcl-2,Cylin D1和Wnt3a表达的阻滞作用。结果：BPEI,LPEI和PAMAM三种材料与miRNA可结合形成稳定的纳米级复合物,在N/P=5时,PC3细胞对BPEI/miRNA-FAM的摄取高于LPEI/miRNA-FAM和PAMAM/miRNA-FAM,具有统计学意义（P<0.01）。BPEI、LPEI和PAMAM都可以携带miRNA-15a和miRNA-16-1进入PC3细胞,并且阻滞PC3细胞中Bcl-2,Cylin D1和Wnt3a蛋白的表达。结论：BPEI,LPEI和PAMAM三种材料包裹miRNA-15a和miRNA-16-1可对激素非依赖型前列腺癌细胞PC3细胞产生抑制作用。并可以有效阻滞PC3细胞中Bcl-2,Cylin D1和Wnt3a蛋白的表达。     

大明胶囊对STZ诱导的1型糖尿病大鼠胰腺microRNA表达谱的影响及其靶点分析

目的 研究大明胶囊(Daming capsule,DM)对链脲佐菌素(streptozocin,STZ)诱导的1型糖尿病大鼠胰腺microRNA (miRNA)表达谱的影响,预测miRNA的靶点并进行分析.方法 将45只成年雄性Sprague-Dawley大鼠随机分为3组:空白组、模型组及DM组.DM组大鼠灌胃给予200 mg/(kg·d)的DM,空白组和模型组给予同等体积的生理盐水,每天2次.2周后,模型组及DM组腹腔注射STZ 65mg/kg建立1型糖尿病模型,注射STZ后DM组大鼠按上述剂量继续给予DM,空白组和模型组给予同等体积的生理盐水,每天2次.STZ注射后3天、7天检测大鼠空腹血糖,并于STZ注射后7天取大鼠胰腺组织.microRNA芯片技术检测各组大鼠胰腺组织miRNA表达,实时荧光定量PCR(real-time PCR,RT-PCR)验证miRNA芯片结果.Targetscan数据库预测miRNA靶点.结果 大鼠腹腔注射STZ 3天和7天后,大鼠空腹血糖值由(6.1±0.6)上升至(21.9±3.1)和(24.6 ±2.4) mmol/L(P ＜0.01).DM组大鼠的空腹血糖为(6.5±0.8) mmol/L,STZ后3天和7天的血糖分别为(14.1±5.1)和(12.4±4.8)mmol/L(P ＜0.01),明显低于同期模型组血糖水平(P＜0.01).在STZ大鼠胰腺,有47个miRNAs表达上调大于2倍,32个miRNAs表达下调大于2倍;DM大鼠胰腺组织,有35个miRNAs表达上调大于2倍,34个miRNAs表达下调大于2倍.其中在STZ大鼠胰腺组织表达上调的21个miRNAs及下调的8个miRNAs的表达被DM逆转;随机选择miR-200b、let-7b和miR-375进行RT-PCR验证的结果显示,这些miRNAs的表达与芯片结果一致;对DM纠正的miRNAs靶点分析发现这些靶点参与了胰腺β细胞胰岛素的生成、分泌和葡萄糖代谢及胰岛细胞凋亡.结论 DM降低了STZ诱导的1型糖尿病大鼠的空腹血糖,并改变了大鼠胰腺组织miRNA表达谱;miRNAs通过调节胰岛素生成、分泌、葡萄糖代谢和胰岛细胞的凋亡参与了DM的降血糖作用.     

miR －139－5p 在胃癌中的表达及意义

目的：分析miR－139－5p在胃癌中的表达及其与临床病理特征的关系。方法采用逆转录实时定量 PCR 方法检测 miR －139－5p 在胃癌及其癌旁正常组织中的表达量,并分析其与临床病理特征关系。此外检测4株胃癌细胞与正常人胃上皮细胞中 miR －139－5p 的表达。结果与癌旁正常组织比较,miR －139－5p 在胃癌组织中表达下调;有无淋巴结转移以及 TNM 分期间 miR －139－5p 的表达差异有统计学意义（P ＜0．05）;而不同性别、年龄、肿瘤大小以及肿瘤分化程度间 miR －139－5p 的表达差异无统计学意义（P ＞0．05）。 miR －139－5p在胃癌细胞株中表达下调。结论miR －139－5p 在胃癌组织和细胞中表达下调,并与胃癌的淋巴结转移及 TNM分期密切相关,其可能参与了胃癌的转移过程。     

High miR-196a and low miR-367 cooperatively correlate with unfavorable prognosis of high-grade glioma

Identification of microRNAs (miRNAs) could be beneficial for the diagnosis and prognosis of glioma. Therefore, we attempted to identify and develop specific miRNAs as prognostic and predictive markers for glioma patients. We compared the expression profiles of 365 miRNAs between 4 glioblastomas (GBMs, WHO grade IV) and 4 anaplastic astrocytomas (AAs, WHO grade III) using miRNA qPCR Array. MiR-196a (P = 0.004, fold change = 289.86) and miR-367 (P = 0.044, fold change = 0.03) were identified as the most up-regulated and down-regulated miRNAs in GBMs compared with AAs, respectively. We subsequently examined miR-196a and miR-367 expression levels in an independent series of 63 gliomas including 50 GBMs and 13 AAs, as well as 10 non-neoplastic brain tissues, and statistically analyzed the associations between miRNA expression and clinicopathological characteristics and survivals of these glioma patients. MiR-196a and miR-367 showed significant increased and decreased expression in high-grade gliomas relative to non-neoplastic brains, as well as in GBMs versus AAs, respectively. Additionally, high-miR-196a and low-miR-367 expression, alone or in combination, statistically correlated with aggressive clinicopathological features of gliomas. Furthermore, overall survivals of glioma patients with high-miR-196a, low-miR-367 and high-miR-196a/low-miR-367 expression tended to be shorter than the corresponding control groups (all P ≤ 0.001). Moreover, multivariate analysis indicated high-miR-196a/low-miR-367 as an independent prognostic indicator for glioma patients (P = 0.005, risk ratio = 1.8). Our results suggested that both high-miR-196a and low-miR-367 expression may be associated with aggressive progression and unfavorable clinical outcome in glioma patients. And combination of high-miR-196a and low-miR-367 expression may be a novel biomarker in identifying a poor prognosis group of high-grade glioma.