12 种 microRNA 在卵巢癌中的表达及临床意义

摘要： 目的 利用公共数据库分析卵巢癌组织中 12 种 microRNA（miR）的表达及其临床意义。方法 筛选在 GSE14407 数据集和 TCGA 数据库中均具有表达信息的 microRNA, 得到 miR-10B、 miR-1244、 miR-622、 miR-21、 miR-503、 Let-7D、 miR-155、 miR-30C、 miR-17、 miR-101-1、 miR-186 以及 miR-770 共 12 种 microRNA, 在这 12 种 microRNA 中寻找在卵巢癌（卵巢癌组）和癌旁组织（癌旁组）中表达有差异的基因。利用在 TCGA 数据库中筛选得 到的 505 例卵巢癌患者数据, 年龄按照中位数分为高年龄（≥59 岁）组和低年龄（＜59 岁）组; 临床分期分为早期组 （Ⅰ期+Ⅱ期）、 晚期组 （Ⅲ期+Ⅳ期） 及未知; 肿瘤分级分为低级别 （G1+G2） 组和高级别 （G3+G4） 组; 病发位置分为单侧 组、 双侧组; 肿瘤残余分为<10 mm 组、 ≥10 mm 组; microRNA 按照表达量的中位数分为高表达组和低表达组, 进行 Kaplan-Meier 单因素生存分析和多因素 Cox 回归分析。结果 癌旁组 Let-7D 和 miR-101-1 表达水平高于卵巢癌 组, 而 miR-155 和 miR-770 表达水平低于卵巢癌组 （均 P < 0.05）。生存分析显示, 年龄≥59 岁、 临床分期 （Ⅲ+ Ⅳ） 期 和 Let-7D 低表达的患者生存情况较差 （P < 0.05）。Let-7D 低表达是卵巢癌患者生存的独立危险因素。结论 Let- 7D 的表达与卵巢癌预后相关, 可能是卵巢癌预后的独立影响指标。     

5-Fluorouracil derivatives: a patent review (2012 – 2014)

Introduction: 5-Fluorouracil (5-FU)-based chemotherapy is the most widely prescribed treatment for gastrointestinal solid tumors, but there are several drawbacks such as toxicities, lack of selectivity and effectiveness as well as the development of resistance that need to be overcome.

Areas covered:In this review, the authors present the latest innovations in 5-FU derivatives or combinations with: i) other chemotherapeutic drugs; ii) novel targeted compounds; iii) radiotherapy; iv) mAbs; v) siRNA strategies; and vi) traditional Chinese medicine extracts. Moreover, advances to overcome or determine 5-FU adverse effects and effectiveness are described. Finally, the authors introduce the ongoing clinical trials and highlight the main challenges to be addressed in the future.

Expert opinion: Although in the past few years there has been a great advancement in the antitumor effectiveness and selectivity of 5-FU-based therapies, it is envisaged that future approaches using ‘omics’ technologies that could determine the tumor heterogeneity may help in identifying additional candidate genes, microRNAs or cytokines involved in both the path mechanisms of 5-FU-related toxicity and its therapeutic efficacy. Moreover, the development of novel targeted 5-FU derivatives or 5-FU-based therapies tailored to individual patients opens up new possibilities in the improvement of the quality of life and survival for those suffering from this devastating disease.     

miR-10b、miR145及miR155在乳腺癌患中的表达及临床意义

目的 探讨microRNA-10b(miR-10b)、microRNA-145(miR-145)及microRNA-155(miR-155)在乳腺癌患者病理组织及血清中的表达水平,进而评价其在乳腺癌诊断中的价值.方法 收集100例正常健康志愿者,150例乳腺癌患者血液和39例乳腺癌患者的癌组织和癌旁组织,提取血液和组织中的microRNA,采用实时荧光定量PCR检测miR-10b、miR-145及miR-155表达水平,分析其与乳腺癌患者临床病理参数之间的相关性,并绘制ROC曲线分析其在乳腺癌诊断中的价值.结果 治疗前乳腺癌患者血清中的miR-10b,miR-145和miR-155表达水平高于对照组(P＜0.05),三者表达水平均与肿瘤转移显著相关,与年龄、组织学特征、ER表达无相关性(P＞0.05)miR-10 b与临床分期显著相关(P＜0.05),miR-10b的ROC曲线下面积为0.936,miR-145的ROC曲线下面积为0.904,miR-155的ROC曲线下面积为0.91.miR-10b,miR-145,miR155在Ⅲ级乳腺癌患者肿瘤组织的表达水平明显高于Ⅰ～Ⅱ级乳腺癌患者癌组织的表达水平(P＜0.05).结论 血清中miR-10 b、miR-145及miR-155有可能作为乳腺癌早期诊断的肿瘤标志物.     

反式-7,8-二羟-9,10-环氧苯并芘诱导恶变细胞中miR-494和miR-22的功能及PTEN的表达

目的分析反式-7,8-二羟-9,10-环氧苯并芘(anti-BPDE)诱导转化人支气管上皮细胞第30代(16HBE-T30)及其前期第15代细胞(16HBE-T15)miR-494及miR-22的表达水平,探讨其在化学致癌过程中的作用及对靶基因PTEN抑癌基因的调控。方法以16HBE-T30和16HBE-T15细胞为实验组,溶剂处理组分别为其相应的对照组细胞(16HBE-N30和16HBE-N15),用茎环结构逆转录引物定量RT-PCR检测四组细胞miR-494及miR-22成熟体表达水平。运用定量RT-PCR和Western blotting分别检测四组细胞PTEN mRNA和蛋白表达水平。运用miRNA前体及抑制剂对miNRA表达进行调控后,再检测PTEN的表达改变,并进行报告基因信号改变、细胞凋亡、软琼脂集落形成率等分析。结果 16HBE-T30中miR-494和miR-22表达水平分别是16HBE-N30的12.6和2.3倍,而16HBE-T15中miR-494、miR-22表达水平较16HBE-N15下降。16HBE-T30中PTEN蛋白表达量低于16HBE-N30。与前体阴性对照组相比,上调的miR-494和miR-22分别将PTEN蛋白的表达量降低至0.8和0.5倍,而降低的miR-494和miR-22分别将PTEN蛋白的表达量上调至1.3和1.5倍。在16HBE-T30细胞中,miR-494和miR-22表达下调均增强caspase-3/7活力,miR-494和miR-22表达上调则减弱其活力。miR-494和miR-22下调能够降低转化细胞的恶性程度。结论 anti-BPDE恶性转化细胞中miR-494和miR-22在转录后水平反向调控靶基因PTEN的表达,这两种miRNA在anti-BPDE致癌过程中发挥类癌基因作用。     

组织与脱落细胞中的microRNA表达量检测指示miR-203在口腔扁平苔藓中的潜在作用

口腔扁平苔藓（OLP）是一种癌前病变和慢性炎症,其损伤具有组织学异质性,多种类型的细胞参与其发生与恶性转化,而其分子机制尚不明确。本研究检测了来自于OLP患者和健康志愿者口腔组织和脱落细胞中miR。96／182／183簇、miR-203、miR-375、miR-769—5p的表达水平,并通过双标图法评价了显著差异表达的miRNA与OLP的相关性。研究发现。相对于志愿者正常组织,miR．203在患者病损区组织中显著高表达,其变化程度明显高于其它miRNA。与此相反,miR-203及miR-96／182／183簇的表达在脱落细胞中则没有显著差异,提示OLP病损区中不同类型细胞对miRNA表达的调控不同,可能与OLP的组织学异质性相关。双标图分析表明miR-203与miR-96／182／183簇的表达量在患者病损区组织中呈正相关。本研究结果表NmiR-203不仅在分子层次显示TOLP的组织学异质性,还可能成为潜在的诊断标志物和治疗药物靶标。     

微RNA-196a和微RNA-210在胰腺癌中的表达及其与生存时间的关系

目的分析微RNA-196a(miR-196a)和miR-210在胰腺癌中的表达,及其与生存时间的关系。方法选择26例胰腺癌患者(A组)、20例良性胰腺病患者(B组)和24例健康受试者(C组)为研究对象。用实时定量荧光聚合酶链式反应法检测3组患者血清中miR-196a和miR-210表达水平。分析3组血清miR-196a和miR-210表达差异,miR-196a和miR-210水平与胰腺癌患者生存时间的关系。结果A,B,C组的miR-196a表达水平分别为(1.79±0.95),(1.24±0.92)和(0.71±0.68),miR-210表达水平(2.32±0.54),(1.25±0.36)和(0.98±0.22),A组的上述指标与B,C组比较,差异均统计学意义(均P<0.05)。在胰腺癌患者中,半年生存率为61.54%,1年生存率为46.15%,2年生存率为3.85%。在胰腺癌患者中,miR-196a高表达组和低表达组的生存时间分别为(7.58±3.43)和(12.38±5.17)个月,miR-210高表达组和低表达组的生存时间分别为(6.92±3.01)和(10.88±4.28)个月,差异均有统计学意义(均P<0.05)。miR-196a的敏感度、特异性及准确率分别为76.92%,93.18%和70.10%,miR-210的敏感度、特异性及准确率分别为68.00%,93.18%和61.18%,联合检测的敏感度、特异性及准确率分别为88.46%,95.45%和83.92%,联合检测的上述指标与miR-196a、miR-210单独检测比较,差异均有统计学意义(均P<0.01)。结论胰腺癌患者血清中特异性的表达miR-196a和miR-210,二者联合检测诊断胰腺癌可提高其诊断准确率,且miR-196a、miR-210表达水平与胰腺癌患者生存时间有一定关系。     

Circulating microRNA 122 in the methionine and choline‐deficient mouse model of non‐alcoholic steatohepatitis

Non‐alcoholic steatohepatitis (NASH) is a progressive form of non‐alcoholic fatty liver disease (NAFLD) and is a major cause of liver cirrhosis and hepatic failure. The methionine choline‐deficient diet (MCD) is a frequently used hepatotoxicity animal model of NASH that induces hepatic transaminase (ALT, AST) elevations and hepatobiliary histological changes similar to those observed in human NASH. Liver‐specific microRNA‐122 (miR‐122) has been shown as a key regulator of cholesterol and fatty acid metabolism in adult liver, and has recently been proposed as a sensitive and specific circulating biomarker of hepatic injury. The purpose of this study was to assess miR‐122 serum levels in mice receiving an MCD diet for 0, 3, 7, 14, 28 and 56 days and compare the performance vs. routine clinical chemistry when benchmarked against the histopathological liver findings. MiR‐122 levels were quantified in serum using RT‐qPCR. Both miR‐122 and ALT/AST levels were significantly elevated in serum at all timepoints. MiR‐122 levels increased on average by 40‐fold after 3 days of initiating the MCD diet, whereas ALT and AST changes were 4.8‐ and 3.3‐fold, respectively. In general, miR‐122 levels remained elevated across all time points, whereas the ALT/AST increases were less robust but correlated with the progressive severity of NASH as assessed by histopathology. In conclusion, serum levels of miR‐122 can potentially be used as a sensitive biomarker for the early detection of hepatotoxicity and can aid in monitoring the extent of NAFLD‐associated liver injury in mouse efficacy models. Copyright © 2013 John Wiley & Sons, Ltd.     

Human Milk Exosomal MicroRNA: Associations with Maternal Overweight/Obesity and Infant Body Composition at 1 Month of Life

Among all the body fluids, breast milk is one of the richest sources of microRNAs (miRNAs). MiRNAs packaged within the milk exosomes are bioavailable to breastfeeding infants. The role of miRNAs in determining infant growth and the impact of maternal overweight/obesity on human milk (HM) miRNAs is poorly understood. The objectives of this study were to examine the impact of maternal overweight/obesity on select miRNAs (miR-148a, miR-30b, miR-29a, miR-29b, miR-let-7a and miR-32) involved in adipogenesis and glucose metabolism and to examine the relationship of these miRNAs with measures of infant body composition in the first 6 months of life. Milk samples were collected from a cohort of 60 mothers (30 normal-weight [NW] and 30 overweight [OW]/obese [OB]) at 1-month and a subset of 48 of these at 3 months of lactation. Relative abundance of miRNA was determined using real-time PCR. The associations between the miRNAs of interest and infant weight and body composition at one, three, and six months were examined after adjusting for infant gestational age, birth weight, and sex. The abundance of miR-148a and miR-30b was lower by 30% and 42%, respectively, in the OW/OB group than in the NW group at 1 month. miR-148a was negatively associated with infant weight, fat mass, and fat free mass, while miR-30b was positively associated with infant weight, percent body fat, and fat mass at 1 month. Maternal obesity is negatively associated with the content of select miRNAs in human milk. An association of specific miRNAs with infant body composition was observed during the first month of life, suggesting a potential role in the infant’s adaptation to enteral nutrition.     

The biology,function, and applications of exosomes in cancer

Exosomes are cell-derived nanovesicles with diameters from 30 to 150 nm, released upon fusion of multivesicular bodies with the cell surface. They can transport nucleic acids, proteins, and lipids for intercellular communication and activate signaling pathways in target cells. In cancers, exosomes may participate in growth and metastasis of tumors by regulating the immune response, blocking the epithelial–mesenchymal transition, and promoting angiogenesis. They are also involved in the development of resistance to chemotherapeutic drugs. Exosomes in liquid biopsies can be used as non-invasive biomarkers for early detection and diagnosis of cancers. Because of their amphipathic structure, exosomes are natural drug delivery vehicles for cancer therapy.