长链非编码RNA及其在妇科肿瘤发生发展中的作用

长链非编码RNA(long non-coding RNA,lnc RNA)曾被认为是基因组转录过程中的"噪音"而被科学界忽视。然而,近年来的许多研究表明lnc RNA并非是"垃圾",而是具有功能的RNA分子,故而受到广泛重视。相比于短链非编码RNA,lnc RNA可谓是"后起之秀",相关研究刚崭露头角,但就目前已有的研究表明,lnc RNA在基因表达调控方面发挥着十分重要的作用,在临床疾病尤其是肿瘤的诊治方面有着不可估量的科学价值。综述lnc RNA的功能、作用机制及其在妇科相关肿瘤发生发展中的意义。     

miRNA和lncRNA在早发性卵巢功能不全中的研究进展

微小RNA(miRNA)和长链非编码RNA(lncRNA)都属于非编码RNA(ncRNAs),它们被认为是一类不编码蛋白质的RNA分子,但能以RNA的形式在表观遗传调控、转录调控以及转录后调控等层面上参与蛋白编码基因调控。许多研究表明miRNA和lncRNA在多囊卵巢综合征、子宫内膜异位症、妇科肿瘤中发挥了重要作用,而近年来的研究发现,miRNA和lncRNA在早发性卵巢功能不全(POI)的发生发展中同样有着重要作用,并且随着深入研究,将有利于逐步揭示POI的发病机制,有助于发现新的生物标志物和靶向治疗位点,为POI的预防、早期诊断和治疗提供新思路。     

piRNA在生殖系统中的研究进展

Piwi蛋白相互作用RNA（piRNA）是一类长度约为30个核苷酸的非编码RNA。目前研究提示piRNA通过与PIWI蛋白家族成员结合后,参与异染色质形成、生殖细胞的发育和维持DNA的完整性。piRNA与小干扰RNA（siRNA）及微小RNA（miRNA）均是近些年发现的非编码小RNA,在功能、分布和分子特征等方面存在着显著不同。piRNA在精子的发生过程中起着重要的生理调节作用,目前对于piRNA的研究逐渐深入,认识也进一步提高。例如,piRNA的结构特点,发挥功能相关作用蛋白,在体内的分布与表达,在生物体内与PIWI发挥作用的方式以及功能。piRNA数据库的建立将对此类小分子RNA的研究有很大的促进作用。     

MicroRNA与多囊卵巢综合征关系的研究进展

Micro RNAs(mi RNA)是一类长度为19~25nt的内源性非编码RNA,主要参与调节基因转录后水平的表达。研究发现mi RNAs在卵巢组织中广泛表达,在卵巢功能调节过程中发挥着重要的作用。近年来,越来越多的证据表明,mi RNAs与多囊卵巢综合征(PCOS)关系密切。mi RNA表达水平的变化与PCOS患者甾体激素调节紊乱、高雄激素血症、胰岛素抵抗及不孕症等主要表现有关。将来,mi RNAs有望成为PCOS疾病诊断和预后评估的潜在生物学标志物,可为PCOS的治疗提供新思路。     

微小RNA在女性生殖调节中的研究进展

微小RNA(miRNA)是含有19~22个核苷酸序列的短链非编码RNA,调控内源性基因的转录后表达。随着miRNA在生殖领域不断被发现,miRNA作为女性生殖系统的重要调控因子越来越受到关注。miRNA通过调节靶基因的转录后水平,调控卵巢颗粒细胞的增殖、分化和凋亡并参与控制类固醇激素的分泌。卵巢功能异常,如多囊卵巢综合征(PCOS)和卵巢早衰(POF)均会发生miRNA调控异常。除此之外,miRNA还通过调节目标基因的表达决定胚胎的整倍体性及胚胎发育的命运。miRNA对于维持子宫内膜容受性非常重要,miRNA的异常表达会导致子宫内膜种植窗的偏移,从而导致胚胎种植失败。对miRNA在生殖调节中作用的了解,有助于发现新的预测体外受精成功率的生物指标和靶向治疗的位点,进一步提高辅助生殖的有效性和安全性。     

长链非编码RNA在子宫内膜癌中的研究进展

长链非编码RNA(long non-coding RNA,lncRNA)在机体多种病理生理过程中均发挥极其重要的调控作用,其功能失调与肿瘤发生发展关系密切,因其数量大、种类多且功能多变,成为近年来多种疾病机制研究的热点。子宫内膜癌(endometrial carcinoma)是女性生殖系统最常见的恶性肿瘤之一,和lncRNA之间关系密切。lncRNA在子宫内膜癌中的研究越来越多,随着研究技术和方法不断进步,探明其对子宫内膜癌的调控机制,将有助于深入揭示子宫内膜癌的发生和发展,也可为该疾病的早期诊断和治疗提供新的思路,并有望作为新的预后相关标记物和药物治疗的靶点。现综述lncRNA在子宫内膜癌中的研究进展。     

长链非编码RNA与妇科肿瘤研究进展

长链非编码RNA(long non-coding RNA,lnc RNA)是一类长度超过200个碱基的RNA分子。由于不参与编码形成蛋白质分子,早期观点普遍认为lnc RNA分子是基因组转录过程中形成的副产物,被定义为不具备生物学功能的转录组"背景噪音"。然而,近年来越来越多的研究发现,在哺乳动物基因组中,lnc RNA分子首先由数千个基因位点转录形成,随后,经过加工的成熟lnc RNA分子在动物胚胎发育、基因表达调节以及疾病发生和发展中起关键性调控作用。目前已有研究证实,在多种人类肿瘤疾病中,部分特异性lnc RNA分子的表达水平在疾病进展的各时期中发生显著了变化。在妇科恶性肿瘤中lnc RNA分子表达变化以及lnc RNA通过上下游靶向分子对疾病发生和发展的影响也有了许多报道。综述这一领域最新的研究进展。     

miRNA在妇科恶性肿瘤发病机制中的研究进展

MicroRNAs(miRNAs)是一种新的内源性非编码RNA(non-coding RNAs,ncRNAs),可通过靶信使RNA(message RNA,mRNA)在转录后水平发挥基因调控作用。近年研究证实,在包括肿瘤在内的多种疾病表达谱中miRNA表达有所改变,提示miRNA可能在不同疾病的发生过程中发挥了某种调节作用。越来越多的研究表明,miRNA与肿瘤的发生、发展、诊断、治疗和预后密切相关。现将miRNA与妇科恶性肿瘤研究方面的进展作一简要综述。     

长链非编码RNA与妇科恶性肿瘤的相关性研究

近年来,曾经被认为是基因组转录"噪音"的长链非编码RNA(long non-coding RNA,lnc RNA)得到了学者们的广泛关注。lnc RNA是一类长度超过200个核苷酸(nt)的RNA,其本身并不编码蛋白,但其与生物信息学功能有着密切的联系,涉及表观遗传调控、细胞周期调控、m RNA降解、肿瘤耐药等多个方面,对肿瘤的发生、侵袭、转移、诊断及预后均起着至关重要的作用。同时,其在与激素以及P53信号通路的作用机制之间也有着一定的关系。学者们借助多种分子生物学研究方法,不断地对lnc RNA功能及其分子调控机制进行解析,发现肿瘤细胞与正常组织来源细胞间lnc RNA表达谱存在明显差异。而且,lnc RNA的异常表达与多种恶性疾病的发生息息相关。lnc RNA与妇科肿瘤相关性的研究为妇科恶性肿瘤的诊断和治疗提供了一个新思路。     

长链非编码RNA在宫颈癌中的作用

宫颈癌是女性最常见的恶性肿瘤之一,我国的发病率和死亡率一直居高不下。人乳头瘤病毒的持续感染是宫颈癌的主要危险因素,但并非唯一因素,遗传和表观遗传因素与宫颈癌的关系不容忽视。近年有研究发现长链非编码RNA (long non-coding RNA,lncRNA)与宫颈癌的发生、发展有关。许多lncRNA可以作为竞争性内源RNA(competing endogenous RNA,ceRNA)广泛参与调控一些关键的信号通路,如Wnt/β-catenin依赖性途径、丝裂原活化蛋白激酶通路、磷脂酰肌醇3激酶/蛋白激酶B通路和Notch信号通路等,从而在宫颈癌的发生、发展中发挥致癌或抑癌作用,因此许多lncRNA已成为宫颈癌新的诊断和预后生物标志物。综述lncRNA的结构与功能,以及lncRNA在宫颈癌中的作用机制。     

The effect of delay in collection and processing on RNA integrity in human placenta: Experiences from rural Africa

This paper examines the relationship between time to processing and RNA quality in placentas collected from women in a field setting in rural Gambia. Placental samples were collected from the villages and transferred to the laboratory. RNA was extracted using Trizol and integrity assessed using the RNA integrity number (RIN). Values were inversely correlated with delay in processing. Expression levels of candidate genes increased with decreasing RIN. Normalising to a housekeeper gene removed this artefact. We propose a cut-off point of 90 min from delivery, after which samples cannot be used for gene expression analysis.     

Aberrant expression of long noncoding RNAs in cumulus cells isolated from PCOS patients

Purpose

To describe the long noncoding RNA (lncRNA) profiles in cumulus cells isolated from polycystic ovary syndrome (PCOS) patients by employing a microarray and in-depth bioinformatics analysis. This information will help us understand the occurrence and development of PCOS.

Methods

In this study, we used a microarray to describe lncRNA profiles in cumulus cells isolated from ten patients (five PCOS and five normal women). Several differentially expressed lncRNAs were chosen to validate the microarray results by quantitative RT-PCR (qRT-PCR). Then, the differentially expressed lncRNAs were classified into three subgroups (HOX loci lncRNA, enhancer-like lncRNA, and lincRNA) to deduce their potential features. Furthermore, a lncRNA/mRNA co-expression network was constructed by using the Cytoscape software (V2.8.3, http://www.cytoscape.org/).

Results

We observed that 623 lncRNAs and 260 messenger RNAs (mRNAs) were significantly up- or down-regulated (≥2-fold change), and these differences could be used to discriminate cumulus cells of PCOS from those of normal patients. Five differentially expressed lncRNAs (XLOC_011402, ENST00000454271, ENST00000433673, ENST00000450294, and ENST00000432431) were selected to validate the microarray results using quantitative RT-PCR (qRT-PCR). The qRT-PCR results were consistent with the microarray data. Further analysis indicated that many differentially expressed lncRNAs were transcribed from chromosome 2 and may act as enhancers to regulate their neighboring protein-coding genes. Forty-three lncRNAs and 29 mRNAs were used to construct the coding-non-coding gene co-expression network. Most pairs positively correlated, and one mRNA correlated with one or more lncRNAs.

Conclusions

Our study is the first to determine genome-wide lncRNA expression patterns in cumulus cells isolated from PCOS patients by microarray. The results show that clusters of lncRNAs were aberrantly expressed in cumulus cells of PCOS patients compared with those of normal women, which revealed that lncRNAs differentially expressed in PCOS and normal women may contribute to the occurrence of PCOS and affect oocyte development.

     

Mechanical stretching induces interleukin-8 gene expression in fetal membranes: a possible role for the initiation of human parturition

Objective: Interleukin-8 (IL-8) is known to play a crucial role in human parturition. We aimed to study the effect of mechanical stretching on the expression of IL-8 in fetal membranes (amniochorion) and decidua. Study design: We examined the expression of IL-8 and its receptor in fetal membranes (amniochorion) and decidua by immunohistochemical staining. Also, we studied the synthesis of IL-8 messenger RNA (mRNA) in the fetal membranes before and after stretching. Results: We found that mechanical stretching within physiological limit increased IL-8 messenger RNA (mRNA) synthesis in fetal membranes and decidua in a time-and load-dependent manner. Application of mechanical force led to markedly increased staining of IL-8 receptor in decidual cells but not in amnion or chorion cells. Conclusion: These results suggested that mechanical stretching was a candidate for one of the signals important for production of IL-8 in fetal membranes and decidua and probably for initiation of a cytokine network at amniochorio-decidual interface through increased expression of IL-8 receptors.     

靶向人类白细胞抗原G1(HLA-G1)的siRNA真核表达载体的构建及表达

目的:构建靶向人类白细胞抗原G1(HLA-G1)的小干扰RNA(siRNA)真核表达载体(pSUPER-HLA-G1),并检测其在滋养细胞系HTR-8/SVneo中的敲减效率。方法:将设计的HLA-G1 siRNA寡聚脱氧核苷酸链与真核表达载体pSUPER连接,构建重组pSUPER-HLA-G1真核表达载体,并用脂质体法将其转染入HTR-8/SVneo细胞系中。pSUPER-HLA-G1质粒转染后采用RT-PCR检测HLA-G1在HTR-8/SVneo细胞中的基因转录情况,Western blotting检测HLA-G1蛋白的表达情况。结果:构建的真核表达载体pSUPER-HLA-G1可在HTR-8/SVneo细胞中表达,HLA-G1 mRNA和其蛋白表达抑制率分别为74.85±7.43%和71.07±6.11%。结论:构建的人HLA-G1 siRNA蛋白真核表达载体pSUPER-HLA-G1有效地沉默了HLA-G1在HTR-8/SVneo滋养细胞中的基因表达及蛋白表达,为今后以HLA-G1为靶点的基因研究奠定了基础。     

RNA结合蛋白Rbpms 2在小鼠卵母细胞及早期胚胎中的表达及意义

目的:检测分析多重剪接RNA结合蛋白2(RNA binding protein with multiple splicing 2,Rbpms2)在小鼠卵母细胞及早期胚胎中的表达及分布情况。方法:运用生物信息学资源分析Rbpms2基因表达的组织分布情况以及Rbpms2蛋白的功能结构域;通过免疫印迹法、免疫组织化学及免疫荧光检测,分析Rbpms2在小鼠卵母细胞及早期胚胎中的表达和分布。结果:生物信息学检索显示Rbpms2 mRNA在卵细胞及受精卵中高表达,其蛋白具有一个RNA识别结构域。免疫印记法证实Rbpms2蛋白在小鼠卵母细胞中高表达,同时免疫组织化学显示阳性信号主要集中在卵母细胞胞质中,免疫荧光显示在早期胚胎中,Rbpms2的信号集中在细胞核中。结论:Rbpms2在小鼠卵母细胞及早期胚胎中呈高水平表达,提示其可能在卵母细胞和早期胚胎的发育过程中发挥重要作用。     

Maternal hepatitis C (HCV) infection and Anti-D immunoglobulin therapy: study testing antibodies,RNA and Genotype of HCV in Baghdad

Introduction: Hepatitis C virus (HCV) infection is a serious health problem. It is a major contributor to end-stage liver disease. Worldwide, 1–8% of all pregnant women were infected. Women with viral hepatitis may be at an increased risk of pregnancy complications. There are several obstetrics intervention acts as risk factors, which are specific to women pertaining the HCV infection; anti-D immunoglobulin (Ig) therapy may be one of them. Our objectives were to estimate the prevalence of HCV antibodies (anti-HCV), RNA, and genotype distribution among women with anti-D Ig therapy.

Materials and methods: A cross sectional study was conducted. A sample of 154 Rhesus negative (Rh???ve) pregnant women regardless of the anti-D Ig therapy was collected. Anti-HCV were tested using third generation enzyme immunoassay (EIA-3) and immunoblot assay (Lia Tek-111), subsequently. In addition, 89 serum samples were subjected to molecular analysis using RT-PCR and DNA enzyme immunoassay (DEIA) method for the detection of HCV-RNA and genotypes.

Results: Anti-HCV, and HCV-RNA seroprevalence were significantly higher (17.1, 35.5%) among women with anti-D Ig than their counter group (6.4, 13.16%), p?=?.038, .018, respectively. Significant direct positive dose response correlation (r?=?0.78, p?=?.005) had been seen between number of anti-D Ig therapy and anti-HCV seropositive rate. Anti-D Ig therapy act as a risk factor (odds ratio (OR)?=?3.01, 95%CI: 1.01–8.9) especially from the third dose onward. Women with anti-D Ig therapy were at higher risk (3.6 times more) of positive HCV-RNA (OR =3.6, 95%CI =1.19–10.837). Genotype HCV-1b showed higher prevalent (52.9%) among the recipients of anti-D Ig therapy while genotype HCV-3a (6.6%) was the lowest.

Conclusions: Our study showed that Anti-D immunoglobulin therapy acts as a risk factor for acquiring HCV infection. Screening for HCV should be recommended for all recipients of anti-D Ig. Not only HCV antibodies but HCV-RNA detection being recommended for the diagnosis of HCV infection.

A brief rational: Pregnant women with HCV infection are at risk of adverse obstetric outcome. Anti-D Ig therapy may be a risk factor for HCV infection. Hence, we conducted a cross sectional study with the objectives to estimate the prevalence of HCV antibodies (anti-HCV), RNA, and genotype distribution among women with anti-D Ig therapy. We found that anti-HCV and HCV-RNA seroprevalence were significantly higher in women with anti-D Ig. In addition, women with anti-D Ig therapy were 3.6 times more at risk of positive HCV-RNA with genotype HCV-1b showed higher prevalence. Therefore, anti-D Ig therapy is a risk factor for acquiring HCV infection and we recommend screening for HCV for all recipients of anti-D Ig. In addition, the diagnosis of HCV infection, should be made with HCV antibodies and HCV-RNA detection.     

siRNA沉默乳腺癌靶基因及递送治疗的研究进展

乳腺癌是最常见的女性恶性肿瘤之一,其治疗仍然存在多药耐药、复发和转移等问题。近年来,小干扰RNA(small interfering RNA,siRNA)在乳腺癌的研究中应用广泛,并有望成为新型的靶向性基因药物。本文检索近十年来siRNA在乳腺癌治疗研究方面的相关文献,着重从siRNA沉默乳腺癌相关的靶基因,siRNA的递送载体以及siRNA联合化疗药物治疗乳腺癌的研究3个方面进行综述,以期为乳腺癌的研究提供新方法和新思路。