核糖核酸干扰技术在移植免疫中的应用前景

核糖核酸干扰（RNA interference，RNAi）是一种由双链RNA诱发的基因沉默现象。自从2001年在哺乳动物培养细胞中通过小型干扰RNA（siRNA）成功诱导了特异性靶基因表达沉默后，又有学者报道用病毒载体系统在原代哺乳动物细胞、干细胞和转基因小鼠上都成功实现了RNAi，更拓宽了这项技术的应用范围。虽然目前RNAi在移植学领域中的相关研究几乎处于空白阶段，但我们可以通过从相关学科RNAi的研究进展中触类旁通，探讨其在移植领域中的应用前景。     

RNA干扰技术在大肠癌治疗中的研究进展

RNA干扰（RNAi）是近几年发展起来的一种阻抑内源基因表达的新方法,是由双链RNA诱发的发生在转录后水平的基因沉默。RNAi技术在逆基因分析及基因治疗中已有广泛的应用。大肠癌的发生是一种多基因、多因素相互作用的结果,目前对于大肠癌基因治疗的实验研究已经有了很大的突破。RNAi与传统的基因治疗手段相比,具有沉默效果好、作用稳定等优点。     

RNAi与肿瘤相关基因及在肿瘤治疗中的研究

RNA干扰（RNA interfcrence,RNAi）,是一种在动植物中广泛存在的通过双链RNA分子在mRNA水平上诱导特异性序列基因沉默的过程,随着对RNAi研究的不断深入,其作用机制正在逐步被阐明;同时作为阻断基因表达的新手段,RNAi技术也日趋完善和成熟。而恶性肿瘤基因学说也越来越得到重视,肿瘤相关基因的研究更加细致深入。本文就RNAi技术的基本原理、肿瘤相关基因的概念以及RNAi技术在肿瘤尤其是泌尿系肿瘤相关基因研究中的应用作一综述。     

RNA干扰在器官移植中的基因治疗靶位点

RNA干扰（RNAi）是指双链RNA（dsRNA）分子在mRNA水平关闭相应序列基因的表达或使其沉默的过程，是一种序列特异性的转录后基因沉默。RNAi广泛存在于生物界中，从低等原核生物到植物、真菌、无脊椎动物，甚至近来在哺乳动物中也发现了此种现象。鉴于RNAi技术的高效性和特异性，目前RNAi技术已广泛应用于基因组学、     

RNA干扰技术及其在泌尿外科研究中的应用

RNA干扰(RNA interfcrence，RNAi)，是一种在动植物中广泛存在的通过双链RNA分子在mRNA水平上诱导特异性序列基因沉默的过程。近年来，随着对RNAi研究的不断深入，其作用机制正在逐步被阐明；同时作为阻断基因表达的新手段，RNAi技术也日趋完善和成熟。以其高效性和特异性，RNAi为泌尿外科研究提供了有力的工具。本文就RNAi技术的有关历史、作用机理及其在泌尿外科研究中的应用作一概述。     

RNA干扰技术及其在泌尿外科研究中的应用

RNA干扰 (RNAinterfcrence ,RNAi) ,是一种在动植物中广泛存在的通过双链RNA分子在mRNA水平上诱导特异性序列基因沉默的过程。近年来 ,随着对RNAi研究的不断深入 ,其作用机制正在逐步被阐明 ;同时作为阻断基因表达的新手段 ,RNAi技术也日趋完善和成熟。以其高效性和特异性 ,RNAi为泌尿外科研究提供了有力的工具。本文就RNAi技术的有关历史、作用机理及其在泌尿外科研究中的应用作一概述     

RNA干扰技术在以VEGF/VEGFR为靶点的膀胱癌治疗研究中的应用前景

RNA干扰（RNAi）是正常生物体内抑制特定基因表达的一种现象，它是指当细胞中导入与内源性mRNA编码区同源的双链RNA（dsRNA），该mRNA发生降解而导致基因沉默的现象。利用RNAi技术干涉VEGF表达，达到抑制肿瘤发展和转移的研究已较为广泛。膀胱癌生长、转移不仅有VEGF表达增多，同时发现VEGFR表达亦增高，成为RNAi的新靶点。现主要综述以VEGF/VEGFR为靶点的RNAi技术在膀胱癌治疗研究中的优势、发展前景以及存在问题与对策。     

RNAi在膀胱癌研究中的应用

RNA干扰(RNA interference,RNAi)是正常生物体内抑制特定基因表达的一种现象,即当细胞中导入与内源性mRNA编码区同源的双链RNA时,该mRNA发生降解而导致基因表达沉默.近来随着RNAi的机制不断被阐明,其作为一种工具在肿瘤的发病机制及治疗方面的应用也日益广泛,本文就RNAi在膀胱癌研究中的进展作一综述.     

RNA干扰技术在动物体内应用的研究进展

RNA干扰（RNA interference,RNAi）技术可以特异地沉默基因,它在细胞水平上的研究已取得了巨大成功,但由于动物模型建立的困难,在动物水平上的研究尚处于起步阶段。本文对近年来RNAi在动物体内研究的应用现状作一综述。     

利用RNA聚合酶Ⅲ依赖的启动子实现条件性RNA干扰

RNA干扰(RNA interference，RNAi)是指由双链RNA引发的同源mRNA特异性降解过程，目前已成为功能基因组学研究的有力工具，并广泛应用于肿瘤基因治疗研究领域。载体介导的RNAi是向细胞内引入小干扰RNA(small interference RNA)的经济、有效的方法，然而由于用来引导siRNA表达的U6、H1等RNA聚合酶Ⅲ依赖的启动子均没有可诱导性或组织特异性，所以在研究特定基因功能和在基因治疗中的应用均受到一定限制。最近研究表明使用顺页式／反式转录调控元件对PolⅢ启动子进行修饰或通过Cre—LoxP系统控制PolⅢ启动子或shRNA的结构可实现可诱导性、组织特异性等条件性RNAi。这些方法的建立不仅拓宽了RNAi技术的应用范围，而且大大提高了基于RNAi技术的肿瘤基因治疗的安全性，具有良好的临床应用前景。     

RNA干扰技术及其在肝癌治疗中的应用

RNA干扰(RNAi)是指生物体内由双链RNA(dsRNA)介导同源mRNA的特异性降解,从而导致基因沉默的现象.随着研究的不断深入,RNAi在肝癌治疗中的应用也取得了积极的进展,显示着巨大的潜力,本文就此予以综述.     

RNA interference therapeutics in organ transplantation: The dawn of a new era

RNA interference (RNAi) is a natural process through which double‐stranded RNA molecules can silence the gene carrying the same code as the particular RNA of interest. In 2006, the discovery of RNAi was awarded the Nobel Prize in Medicine and its success has accumulated since. Gene silencing through RNAi has been used successfully in a broad range of diseases, and, more recently, this technique has gained interest in the field of organ transplantation. Here, genes related to ischemia‐reperfusion injury (IRI) or graft rejection may be silenced to improve organ quality after transplantation. Several strategies have been used to deliver siRNA, and pretransplant machine perfusion presents a unique opportunity to deliver siRNA to the target organ during ex situ preservation. In this review, the potential of RNAi in the field of organ transplantation will be discussed. A brief overview on the discovery of RNAi, its mechanism, and limitations are included. In addition, studies using RNAi to target genes related to IRI in liver, kidney, lung, and heart transplantation are discussed.     

RNA interference: a practical approach

BACKGROUND: Few new molecular biology techniques have advanced to find practical application as rapidly as RNA interference (RNAi). RNAi denotes the highly specific posttranslational silencing of gene expression that occurs in response to the introduction of double-stranded RNA into a cell. The purpose of this review is to present practical guidelines for designing and executing RNAi experiments. MATERIALS AND METHODS: Review of recent literature. RESULT AND CONCLUSION: We summarize the mechanisms underlying RNAi in mammalian cells and focus on practical advice for investigators conducting RNAi experiments. We suggest criteria to help select a suitable target gene sequence, define the structural characteristics of effective siRNAs, discuss transfection strategies, and describe experimental design, including important control methods. RNAi represents a powerful tool for determining the functions of specific genes.     

人CCL5基因RNA干扰慢病毒载体的构建

目的 构建人CCL5基因RNA干扰(RNAi)慢病毒载体.方法 根据人CCL5基因信息,构建4个携带RNAi序列的pGCSIL-GFP质粒,与pHelper 1.0、Helper 2.0质粒一起利用293T细胞进行慢病毒包装.用CCL5 RNAi慢病毒载体感染人宫颈癌细胞(Hela),使用RT-PCR方法验证其干扰效率.结果 4个靶点中有3个靶点(a1、a2、a3)在Hela细胞上对CCL5基因的表达都有非常显著的敲减效果,敲减效率均达到95%以上.结论 构建的CCL5 RNAi慢病毒载体在Hela细胞中有较高的敲减效率,提示RNAi技术能够使细胞CCL5基因沉默.     

RNA干扰在破骨细胞分子调控应用的研究进展

骨质疏松症及其相关骨疾病由成骨细胞和破骨细胞比例失调造成的,导致骨折,致使平均寿命预期降低,迫切需要新的治疗方法。RNA干扰(RNA interference,RNAi)是一种序列特异性转录后基因沉默技术,可用于调控蛋白质的表达,目前被用作阐明疾病相关基因的工具。破骨细胞是一类由骨髓造血干细胞来源的多核巨细胞,其主要作用是介导骨组织的吸收。目前对破骨细胞生物学特性、分化调控机制以及相关通路的研究比较多,但是关于RNAi在破骨细胞分子调控方面的研究尚缺乏。本文就RNAi在破骨细胞分子调控应用方面的研究作相关综述分析,介绍RNAi技术敲除过度骨吸收的多种分子靶标,为科研使用RNAi对破骨细胞的研究提供参考,以期更好地发展靶向的治疗策略。     

RNAi 在大肠癌基因治疗中的应用策略和展望

目的 探讨RNA干扰（RNA interference，RNAi）技术在大肠癌基因治疗研究中的应用。方法复习近几年的相关文献并进行综述。结果RNAi能够高效特异地沉默同源基因表达，可在大肠癌发生、发展多个环节发挥重要作用。结论RNAi技术为大肠癌基因治疗研究开辟了新的途径。     

Induction of accommodation model by combined RNA interference targeting 1,3-galactosyltransferase gene and low-dose GS-IB4 lectin in vitro

OBJECTIVE: This study sought to mimic the interaction of xenograft endothelial cells and human serum in vitro after successfully silencing the expression of porcine alpha1,3-galactosyltransferase (alpha1,3GT) gene by RNA interference (RNAi), and to investigate the possibility of inducing accommodation in vitro by stimulation of alpha-Gal-specific binding lectin, Griffonia simplicifolia isolectin B4 (GS-IB4) and RNAi. MATERIALS AND METHODS: Various alpha-Gal expression patterns on a pig endothelial cell immortalized line (PED) was achieved by serial doses of small interfering RNA (siRNA) targeting porcinc alpha1,3GT gene. alpha1,3GT-siRNA transfected PEDs were exposed to increasing doses of GS-IB4 lectin (0.5, 2, and 8 microg/mL) for 4 hours before incubation with normal human serum (NHS). Accommodation phenomenon of PEDs in NHS was observed by 51Cr release and antibody/complement binding assays. RESULTS: With combined RNAi and low-dose GS-IB4 stimulation, PEDs remarkably inhibited complement-mediated cytotoxicity, which showed a better protective effect than using RNAi alone. At a concentration of 2 mug/mL, GS-IB4 exhibited the maximum protective effect. The expression of E-selectin on alpha1,3GT-siRNA transfected PEDs did not differ from that on parental PEDs with heat-inactivated NHS (HINHS) stimulation. Combined with GS-IB4 stimulation, however, it inhibited expression of E-selectin, which was GS-IB4 dose dependent, resulting in mean fluorescence intensity values of 98.5, 42.0, and 36.3 at 0.5, 2, and 8 microg/mL. The mRNA expression of the protective gene HO-1 was significantly up-regulated after treatment with RNAi and low-dose of GS-IB4. CONCLUSIONS: Combined RNAi and low-dose GS-IB4 induced pig endothelial cell accommodation in vitro. The level of alpha-Gal expression played an important role in the induction of accommodation.     

体外构建的小片段发夹RNA对肿瘤细胞端粒酶基因表达的干扰

目的:探讨针对端粒酶hTERT基因的小片段发夹RNA对人肿瘤细胞（HeLa）端粒酶基因的干扰及对肿瘤端粒酶的抑制作用。方法:体外合成制备shRNA，并用磷酸钙转染法转染HeLa细胞，分别用TRAP－银染法和PCR－EIA法检测端粒酶活性。结果:将制备的46个碱基的小片段发夹RNA转染HeLa细胞后端粒酶活性明显下降。结论:shRNA对肿瘤端粒酶hTERT基因表达有明显的干扰作用, 可望为临床开展对其他肿瘤端粒酶基因干扰抑制的实验研究奠定基础。     

慢病毒介导的RNA干扰对大鼠神经元PKCγ mRNA及蛋白表达的影响

目的 探讨慢病毒介导的RNA干扰(RNAi)对大鼠神经元PKCymRNA及蛋白表达的影响.方法 孕16 d的SD大鼠,原代培养大鼠胚胎皮层神经元,随机分为3组,每组6孔,对照组(C组):不行任何处理;阴性对照组(NC组):每孔加入阴性对照慢病毒3×105个;RNAi组:每孔中加入编码PKCγ shRNA的慢病毒载体(LV-PKCγ shRNA)颗粒3×105个.5 d后采用RT-PCR和Western blot法检测PKCγ mRNA和蛋白的表达水平,并计算干扰效率.结果 与C组比较,RNAi组大鼠神经元PKCγ mRNA及其蛋白表达水平均降低(P<0.05),NC组上述指标差异无统计学意义(P>0.05).基因干扰效率99.3%,蛋白干扰效率85.2%.结论 慢病毒介导的RNAi可有效地下调大鼠原代神经元PKCγ基因及其蛋白的表达.     

Ubiquitin-specific protease 2 regulates hepatic gluconeogenesis and diurnal glucose metabolism through 11β-hydroxysteroid dehydrogenase 1

Hepatic gluconeogenesis is important for maintaining steady blood glucose levels during starvation and through light/dark cycles. The regulatory network that transduces hormonal and circadian signals serves to integrate these physiological cues and adjust glucose synthesis and secretion by the liver. In this study, we identified ubiquitin-specific protease 2 (USP2) as an inducible regulator of hepatic gluconeogenesis that responds to nutritional status and clock. Adenoviral-mediated expression of USP2 in the liver promotes hepatic glucose production and exacerbates glucose intolerance in diet-induced obese mice. In contrast, in vivo RNA interference (RNAi) knockdown of this factor improves systemic glycemic control. USP2 is a target gene of peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), a coactivator that integrates clock and energy metabolism, and is required for maintaining diurnal glucose homeostasis during restricted feeding. At the mechanistic level, USP2 regulates hepatic glucose metabolism through its induction of 11β-hydroxysteroid dehydrogenase 1 (HSD1) and glucocorticoid signaling in the liver. Pharmacological inhibition and liver-specific RNAi knockdown of HSD1 significantly impair the stimulation of hepatic gluconeogenesis by USP2. Together, these studies delineate a novel pathway that links hormonal and circadian signals to gluconeogenesis and glucose homeostasis.