2021年表观遗传学技术资讯 --- 第二期 RNA pull down技术及最新成果（20210101-0124）_

2021年表观遗传学技术资讯 --- 第二期 RNA pull down技术及最新成果（20210101-0124）

发布时间：2021-01-27| BY ：伯信官网

本期给大家带来表观遗传学研究中使用RNA pull down技术的最新研究成果；本期从PubMed中检索收集最新发布的使用RNA pull down技术的文献共计78篇，下面我们一起来看看，最近的研究中哪些技术路线选用了RNA pull down技术以及对结果分析起到什么样的作用。检索式：(RNA pull down[Title/Abstract])

1、miR-552-3p调节FXR和LXR的转录活性以改善肝糖脂代谢异常。

标题：miR-552-3p modulates transcriptional activities of FXR and LXR to ameliorate hepatic glycolipid metabolism disorder.

杂志：J Hepatol

影响因子：20.582

通讯作者：陈静，金仁（中国科学院上海药物研究所药物安全评价研究中心）和谢庆（上海交通大学医学院附属瑞金医院传染科）

miRNA的核定位已经发现了十多年了，但是miRNA在细胞核中的确切机制和功能仍未完全阐明。研究者发现，核内miR-552-3p在基因转录中具有抑制作用，以及其特有的aggtca样序列，即NR1亚家族的顺式元件,他们旨在探讨其对NR1s的潜在作用以及在改善NR1中的可能应用肝糖脂代谢。研究后发现MiR-552-3p在体外可抑制代谢基因的表达，对体内糖脂代谢具有有益作用。核内miR-552-3p主要调节LXRα和FXR途径。这是通过与AGGTCA的互补序列结合来调节LXRα和FXR的转录活性而实现的。此外，LXRα和FXR配体可以恢复miR-552-3p对基因表达和糖脂代谢的影响。研究结果表明miR-552-3p调节LXRα和FXR的机制揭示了miRNA介导的基因调控的新方法。此外，miR-552-3p的体内有益作用和临床意义提示其可能成为治疗糖脂代谢疾病的潜在靶点。

文中运用RNA Pulldown 技术来验证结合推测。

2、环状RNA circSDHC充当miR-127-3p的海绵，通过CDKN3 / E2F1轴促进肾细胞癌的增殖和转移。

标题：Circular RNA circSDHC serves as a sponge for miR-127-3p to promote the proliferation and metastasis of renal cell carcinoma via the CDKN3/E2F1 axis.

杂志：Mol Cancer

影响因子：15.302

通讯作者：张嘉兴（中山大学附属第一医院肿瘤科）、陈伟和罗俊航（中山大学附属第一医院泌尿外科）

越来越多的证据表明，环状RNA（circRNA）在癌症的发生和发展中具有重要的调控作用。但是，circRNA在肾细胞癌（RCC）中的表达模式和生物学功能仍然难以捉摸。通过研究发现，临床上circSDHC的高表达与RCC患者的TNM分期晚期和生存不良有关。另外，circSDHC在体内和体外均促进肿瘤细胞的增殖和侵袭。研究者对circSDHC在RCC中作用的潜在机制的分析表明，它与miR-127-3p竞争性结合并阻止其抑制下游基因CDKN3和E2F1途径，从而导致RCC恶性进展。此外，敲除circSDHC会导致CDKN3表达降低和E2F1途径抑制，这可以通过用miR-127-3p抑制剂治疗来挽救。该研究数据首次表明circSDHC / miR-127-3p / CDKN3 / E2F1轴在RCC进展中起着至关重要的作用。circSDHC有望成为RCC患者的新治疗靶点。

文中运用RNA Pulldown 技术来确定circSDHC, miR-127-3p 及其靶基因之间的相互作用。

3、来源于低氧胶质瘤干细胞样细胞的细胞外囊泡通过传递miR-30b-3p对胶质母细胞瘤产生替莫唑胺耐药性

标题：Extracellular vesicles derived from hypoxic glioma stem-like cells confer temozolomide resistance on glioblastoma by delivering miR-30b-3p

杂志：Theranostics

影响因子：8.579

通讯作者：尤永平、张俊霞（南京医科大学附属第一医院神经外科）

胶质母细胞瘤(Glioblastoma，GBM)是成人最常见的原发性脑恶性肿瘤。尽管目前采用了最大限度的手术切除、放疗和化疗，新诊断的GBM患者的预后仍然很差，中位生存期不到两年。替莫唑胺(TMZ)是治疗GBM的一线化疗药物。然而，胶质瘤干细胞(GSCs)，一种能够自我更新的癌细胞亚群，由于其治疗耐药性而限制了GBMs的有效治疗。GSC的产生和维护需要一个支持性的利基环境。周围的基质细胞，如反应性星形胶质细胞、免疫细胞和内皮细胞，被认为是GSC利基的组成部分。缺氧被认为是几种恶性肿瘤对放化疗耐药的诱因。缺氧诱导因子(HIF)蛋白的稳定和激活，特别是HIF1α和HIF2α被认为是细胞对缺氧的重要反应。HIF1α和HIF2α在恶性肿瘤的发生和发展中起着基因激活剂的作用。神经干细胞通常存在于低氧微环境中，并假设低氧支持其未分化状态和治疗抵抗的维持。然而，低氧维持GSC茎性的潜在机制尚不清楚。肿瘤细胞排出的细胞外囊泡(EV)是细胞间通讯的重要媒介，因此影响肿瘤的发生。EV介导的肿瘤细胞间的通信在该领域得到了越来越多的关注，许多研究都集中在肿瘤或基质细胞来源的EVS的影响上。EVS也被证明参与了微环境的重新编程。然而，从GSC释放的细胞外囊泡的作用却鲜为人知。因此研究者试图研究EV介导的GSCs在不同微环境中的通讯。研究者假设维持在低氧环境中的GSCs会释放含有特定成分的EVS，这些成分可能会对GBM产生治疗抗性。

胶质瘤干细胞样细胞（GSC）有助于胶质瘤中替莫唑胺（TMZ）的耐药性，尽管其机制尚未阐明。采用体外功能实验(集落形成实验、流式细胞仪分析、TUNEL法)检测缺氧GSC胞外囊泡(EV)对胶质母细胞瘤(GBM)细胞TMZ耐药性的影响。采用RNA测序和定量逆转录聚合酶链反应(QRT-PCR)方法鉴定缺氧性EV中具有功能的miRNA。染色质免疫沉淀分析HIF1、α和STAT3对miRNAs的转录调控。RIP和RNA下拉试验用于验证hnRNPA2B1介导的miRNA包装到EVS中。通过体内实验和临床标本分析，证实了缺氧GSCEV miR-30b-3p的功能。研究结果表明低氧GSC来源的EVS对GBM化疗耐药的影响大于常氧GSC来源的EVS。MiRNA图谱显示，缺氧GSCs的EVS中miR-30b-3p表达显著上调。此外，HIF1α和STAT3在转录水平诱导miR-30b-3p的表达。RNA免疫沉淀和RNA下拉实验结果表明，miR-30b-3p与hnRNPA2B1结合促进了miR-30b-3p向EVS的转移。EV包装的miR-30b-3p(EV-miR-30b-3p)可直接靶向RHOB，在体内外均能减少细胞凋亡，促进细胞增殖。研究结果证明了脑脊液中的miR-30b-3p可能成为预测TMZ耐药性的潜在生物标志物。研究者得到的结论是靶向EV-miR-30b-3p可为GBM提供一种潜在的治疗策略。

文中运用RNA Pulldown 技术来验证hnRNPA2B1介导的miRNA包装到EVS中。

参考文献列表

序号	标题	杂志	影响因子
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7	Long noncoding RNA LINC00671 exacerbates osteoarthritis by promoting ONECUT2-mediated Smurf2 expression and extracellular matrix degradation.	Int Immunopharmacol	3.943
8	Circ_0020397 regulates the viability of vascular smooth muscle cells by up-regulating GREM1 expression via miR-502-5p in intracranial aneurysm.	Life Sci	3.647
9	Long noncoding RNA ATB promotes ovarian cancer tumorigenesis by mediating histone H3 lysine 27 trimethylation through binding to EZH2.	J Cell Mol Med	4.486
10	LncRNA HOTAIR Participates in Microglia Activation and Inflammatory Factor Release by Regulating the Ubiquitination of MYD88 in Traumatic Brain Injury.	J Mol Neurosci	2.678
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