药物转运体表观遗传调控机制的研究进展

药物转运体在体内药物吸收、分布和排泄过程中发挥着重要的作用。转运体在各组织器官的分布和表达受到表观遗传修饰调控,导致某些药物体内处置过程出现明显的个体差异。随着表观遗传学的发展,基于表观遗传修饰（如DNA甲基化、组蛋白修饰、microRNA干预等）调控药物转运体的相关研究越来越多。对表观遗传调控药物转运体研究进行综述。     

Pharmacoepigenetics: an element of personalized therapy?

Introduction: Epigenetics is a rapidly growing field describing heritable alterations in gene expression that do not involve DNA sequence variations. Advances in epigenetics and epigenomics have influenced pharmacology, leading to the development of a new specialty, pharmacoepigenetics, the study of the epigenetic basis for the individual variation in drug response.

Areas covered: We present an overview of the major epigenetic mechanisms and their effects on the expression of drug metabolizing enzymes and drug transporters, as well as the epigenetic status of drug protein targets affecting therapy response. Recent advances in the development of pharmacoepigenetic biomarkers and epidrugs are also discussed.

Expert opinion: There is growing evidence that pharmacoepigenetics has the potential to become an important element of personalized medicine. Epigenetic modifications influence drug response, but they can also be modulated by drugs. Moreover, they can be monitored not only in the affected tissue, but also in body fluids. Nevertheless, there are very few examples of epigenetic biomarkers implemented in the clinical setting. Explanation of the interplay between genomic and epigenomic changes will contribute to the personalized medicine approach. Ultimately, both genetic biomarkers and epigenetic mechanisms should be taken into consideration in predicting drug response in the course of successful personalized therapy.     

苦参碱通过抑制microRNA-192调控顺铂耐受肺腺癌A549细胞对顺铂的敏感性

目的探讨苦参碱通过调控microRNA影响顺铂耐受A549(A549/DDP)细胞对顺铂的敏感性。方法采用生物信息学手段对GEO数据库中的相关microRNA芯片数据(访问编号GSE43249)进行数据挖掘,筛选出与A549细胞顺铂敏感性相关的microRNA。体外培养对顺铂敏感的A549细胞及A549/DDP,使用实时荧光定量PCR(qRT-PCR)法在体外培养的细胞中验证生物信息学的结果,并考察苦参碱对A549/DDP细胞中microRNA水平的影响。通过转染micro RNA-192(mi R-192)模拟序列上调A549/DDP细胞的mi R-192水平。CCK-8细胞活力实验考察苦参碱处理及(或)mi R-192上调对A549/DDP细胞顺铂敏感性的影响。结果芯片数据挖掘显示与顺铂敏感的A549细胞相比,A549/DDP细胞中共有11个microRNA存在显著变化(P0.05),其中mi R-192及mi R-194变化在体外培养的细胞中得到了验证。与未经处理的A549/DDP细胞相比,苦参碱处理后的A549/DDP细胞中mi R-192水平显著降低(P0.05)。苦参碱处理可以增加A549/DDP细胞对顺铂的敏感性,miR-192的上调可以消除这种效果。上调miR-192水平可诱导A549细胞的顺铂耐受。结论苦参碱可能通过抑制mi R-192调控A549对顺铂的敏感性。     

MiR-663a Inhibits Radiation-Induced Epithelium-to-Mesenchymal Transition by Targeting TGF-β1

Objective miR-663 a has been reported to be downregulated by X-ray irradiation and participates in radiation-induced bystander effect via TGF-β1. The goal of this study was to explore the role of mi R-663 a during radiation-induced Epithelium-to-mesenchymal transition(EMT).Methods TGF-β1 or IR was used to induce EMT. After mi R-663 a transfection, cell migration and cell morphological changes were detected and the expression levels of mi R-663 a, TGF-β1, and EMT-related factors were quantified.R...     

海藻糖低温保存血小板miRNA差异表达分析

目的 探讨海藻糖低温保存血小板过程中miRNA的变化.方法 收集3份来自相同血型无偿献血员的单采血小板,少量混匀后,平均分为2组.对照组:新鲜血小板组(22℃,100％血浆储存的新鲜血小板2h);实验组:海藻糖低温保存血小板组(10℃,70％海藻糖保存液储存5d);采用高通量测序方法对两组样品进行了Small RNA测序,进行生物信息分析,主要包括已知miRNA鉴定、miRNA差异表达分析及富集分析.结果 2组样品经Small RNA高通量测序,平均每个样品获得了11兆净读值(Clean reads),经过生物信息分析,分别获得了821个已知miRNA.通过差异表达分析,在已知的miRNA中共有460个差异表达miRNA,其中194个miRNA表达上调,266个miRNA表达下调,其中显著差异表达已知miRNA共44个,分别在钙离子通路、MAPK通路、凋亡路径、轴突导向等储存损伤相关路径中富集程度较高.相对于对照组,实验组中hsa-miR-4449、hsa-miR-1296表达显著上调(P ＜0.01,且差异倍数＞2),hsa-miR-665表达被抑制.结论 血小板经过海藻糖低温保存后的miRNA呈现出特征性表达变化,提示miRNA活动参与了低温时海藻糖对血小板储存损伤的保护作用.     

MicroRNA-539靶向膜相关的鸟苷酸激酶蛋白1抑制甲状腺癌细胞的侵袭与迁移

目的 观察microRNA-539（miR-539）的表达对甲状腺癌细胞侵袭与迁移的影响,探究miR-539与膜相关的鸟苷酸激酶蛋白1（CARMA1）在甲状腺癌中相互作用的机制。方法 构建miR-539与CARMA1高表达及低表达的细胞模型后通过transwell及MTT法检测两种不同的甲状腺癌细胞的侵袭、迁移及增殖能力。Western blot及RT-PCR检测细胞中蛋白及mRNA表达。结果  miR-539抑制甲状腺癌细胞的侵袭与迁移。通过荧光素酶报告检测发现miR-539通过靶向CARMA1的3’-UTR端从而起到抑制甲状腺癌中CARMA1的作用。进一步的研究证实,CARMA1可显著促进甲状腺癌细胞的侵袭与迁移。而调节CARMA1的表达,miR-539随之变化。研究还发现,与对照组比较,甲状腺癌中miR-539的表达量降低而CARMA1的表达量增加。结论  miR-539靶向CARMA1抑制甲状腺癌细胞的侵袭与迁移。

     

microRNA定量检测方法研究现状

MicroRNA(miRNA)是一类由18～24 nt核酸构成的非编码内源性小分子RNA,主要参与真核生物的细胞分化、增殖与凋亡等转录后水平的基因表达调控.研究发现,miRNA与肿瘤、代谢调控、疾病发生及诊断等方面有密切的联系,因此准确且灵敏地进行miRNA的定量检测是深入研究miRNA功能的前提.目前,miRNA定量检测原理主要基于核酸杂交或扩增,包括Northern印迹、微阵列芯片、实时定量PCR、滚环扩增等.本文通过对传统miRNA检测方法、高灵敏度新型miRNA检测方法进行阐述与分析,为筛选合适的miRNA定量检测方法提供参考.