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2021年表觀遺傳學技術資訊 --- 第二期 RNA pull down技術及最新成果（20210101-0124）

發布時間：2021-01-27| BY ：伯信官網

本期給大家帶來表觀遺傳學研究中使用RNA pull down技術的最新研究成果；本期從PubMed中檢索收集最新發布的使用RNA pull down技術的文獻共計78篇，下面我們一起來看看，最近的研究中哪些技術路線選用了RNA pull down技術以及對結果分析起到什么樣的作用。檢索式：(RNA pull down[Title/Abstract])

1、miR-552-3p調節FXR和LXR的轉錄活性以改善肝糖脂代謝異常。

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

雜志：J Hepatol

影響因子：20.582

通訊作者：陳靜，金仁（中國科學院上海藥物研究所藥物安全評價研究中心）和謝慶（上海交通大學醫學院附屬瑞金醫院傳染科）

miRNA的核定位已經發現了十多年了，但是miRNA在細胞核中的確切機制和功能仍未完全闡明。研究者發現，核內miR-552-3p在基因轉錄中具有抑制作用，以及其特有的aggtca樣序列，即NR1亞家族的順式元件,他們旨在探討其對NR1s的潛在作用以及在改善NR1中的可能應用肝糖脂代謝。研究后發現MiR-552-3p在體外可抑制代謝基因的表達，對體內糖脂代謝具有有益作用。核內miR-552-3p主要調節LXRα和FXR途徑。這是通過與AGGTCA的互補序列結合來調節LXRα和FXR的轉錄活性而實現的。此外，LXRα和FXR配體可以恢復miR-552-3p對基因表達和糖脂代謝的影響。研究結果表明miR-552-3p調節LXRα和FXR的機制揭示了miRNA介導的基因調控的新方法。此外，miR-552-3p的體內有益作用和臨床意義提示其可能成為治療糖脂代謝疾病的潛在靶點。

文中運用RNA Pulldown 技術來驗證結合推測。

2、環狀RNA circSDHC充當miR-127-3p的海綿，通過CDKN3 / E2F1軸促進腎細胞癌的增殖和轉移。

標題：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.

雜志：Mol Cancer

影響因子：15.302

通訊作者：張嘉興（中山大學附屬第一醫院腫瘤科）、陳偉和羅俊航（中山大學附屬第一醫院泌尿外科）

越來越多的證據表明，環狀RNA（circRNA）在癌癥的發生和發展中具有重要的調控作用。但是，circRNA在腎細胞癌（RCC）中的表達模式和生物學功能仍然難以捉摸。通過研究發現，臨床上circSDHC的高表達與RCC患者的TNM分期晚期和生存不良有關。另外，circSDHC在體內和體外均促進腫瘤細胞的增殖和侵襲。研究者對circSDHC在RCC中作用的潛在機制的分析表明，它與miR-127-3p競爭性結合并阻止其抑制下游基因CDKN3和E2F1途徑，從而導致RCC惡性進展。此外，敲除circSDHC會導致CDKN3表達降低和E2F1途徑抑制，這可以通過用miR-127-3p抑制劑治療來挽救。該研究數據首次表明circSDHC / miR-127-3p / CDKN3 / E2F1軸在RCC進展中起著至關重要的作用。circSDHC有望成為RCC患者的新治療靶點。

文中運用RNA Pulldown 技術來確定circSDHC, miR-127-3p 及其靶基因之間的相互作用。

3、來源于低氧膠質瘤干細胞樣細胞的細胞外囊泡通過傳遞miR-30b-3p對膠質母細胞瘤產生替莫唑胺耐藥性

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

雜志：Theranostics

影響因子：8.579

通訊作者：尤永平、張俊霞（南京醫科大學附屬第一醫院神經外科）

膠質母細胞瘤(Glioblastoma，GBM)是成人最常見的原發性腦惡性腫瘤。盡管目前采用了最大限度的手術切除、放療和化療，新診斷的GBM患者的預后仍然很差，中位生存期不到兩年。替莫唑胺(TMZ)是治療GBM的一線化療藥物。然而，膠質瘤干細胞(GSCs)，一種能夠自我更新的癌細胞亞群，由于其治療耐藥性而限制了GBMs的有效治療。GSC的產生和維護需要一個支持性的利基環境。周圍的基質細胞，如反應性星形膠質細胞、免疫細胞和內皮細胞，被認為是GSC利基的組成部分。缺氧被認為是幾種惡性腫瘤對放化療耐藥的誘因。缺氧誘導因子(HIF)蛋白的穩定和激活，特別是HIF1α和HIF2α被認為是細胞對缺氧的重要反應。HIF1α和HIF2α在惡性腫瘤的發生和發展中起著基因激活劑的作用。神經干細胞通常存在于低氧微環境中，并假設低氧支持其未分化狀態和治療抵抗的維持。然而，低氧維持GSC莖性的潛在機制尚不清楚。腫瘤細胞排出的細胞外囊泡(EV)是細胞間通訊的重要媒介，因此影響腫瘤的發生。EV介導的腫瘤細胞間的通信在該領域得到了越來越多的關注，許多研究都集中在腫瘤或基質細胞來源的EVS的影響上。EVS也被證明參與了微環境的重新編程。然而，從GSC釋放的細胞外囊泡的作用卻鮮為人知。因此研究者試圖研究EV介導的GSCs在不同微環境中的通訊。研究者假設維持在低氧環境中的GSCs會釋放含有特定成分的EVS，這些成分可能會對GBM產生治療抗性。

膠質瘤干細胞樣細胞（GSC）有助于膠質瘤中替莫唑胺（TMZ）的耐藥性，盡管其機制尚未闡明。采用體外功能實驗(集落形成實驗、流式細胞儀分析、TUNEL法)檢測缺氧GSC胞外囊泡(EV)對膠質母細胞瘤(GBM)細胞TMZ耐藥性的影響。采用RNA測序和定量逆轉錄聚合酶鏈反應(QRT-PCR)方法鑒定缺氧性EV中具有功能的miRNA。染色質免疫沉淀分析HIF1、α和STAT3對miRNAs的轉錄調控。RIP和RNA下拉試驗用于驗證hnRNPA2B1介導的miRNA包裝到EVS中。通過體內實驗和臨床標本分析，證實了缺氧GSCEV miR-30b-3p的功能。研究結果表明低氧GSC來源的EVS對GBM化療耐藥的影響大于常氧GSC來源的EVS。MiRNA圖譜顯示，缺氧GSCs的EVS中miR-30b-3p表達顯著上調。此外，HIF1α和STAT3在轉錄水平誘導miR-30b-3p的表達。RNA免疫沉淀和RNA下拉實驗結果表明，miR-30b-3p與hnRNPA2B1結合促進了miR-30b-3p向EVS的轉移。EV包裝的miR-30b-3p(EV-miR-30b-3p)可直接靶向RHOB，在體內外均能減少細胞凋亡，促進細胞增殖。研究結果證明了腦脊液中的miR-30b-3p可能成為預測TMZ耐藥性的潛在生物標志物。研究者得到的結論是靶向EV-miR-30b-3p可為GBM提供一種潛在的治療策略。

文中運用RNA Pulldown 技術來驗證hnRNPA2B1介導的miRNA包裝到EVS中。

參考文獻列表

序號	標題	雜志	影響因子
1	miR-552-3p modulates transcriptional activities of FXR and LXR to ameliorate hepatic glycolipid metabolism disorder.	J Hepatol	20.582
2	Hsa_circ_0002060 Knockdown Ameliorates Osteoporosis by Targeting MiR-198-5p.	Biol Pharm Bull	1.863
3	CircSFMBT2 facilitates vascular smooth muscle cell proliferation by targeting miR-331-3p/HDAC5.	Life Sci	3.647
4	Long noncoding RNA SNHG14 regulates ox-LDL-induced atherosclerosis cell proliferation and apoptosis by targeting miR-186-5p/WIPF2 axis.	Hum Exp Toxicol	2.067
5	Long Noncoding RNA SNHG16 Facilitates Abdominal Aortic Aneurysm Progression through the miR-106b-5p/STAT3 Feedback Loop.	J Atheroscler Thromb	3.876
6	Long non-coding RNA CCAT1 promotes non-small cell lung cancer progression by regulating the miR-216a-5p/RAP2B axis.	Exp Biol Med (Maywood)	3.139
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
11	Circular RNA circDLC1 inhibits MMP1-mediated liver cancer progression via interaction with HuR.	Theranostics	8.579
12	PfKsgA1 functions as a transcription initiation factor and interacts with the N-terminal region of the mitochondrial RNA polymerase of Plasmodium falciparum.	Int J Parasitol	3.53
13	MicroRNA-150 affects endoplasmic reticulum stress via MALAT1-miR-150 axis-mediated NF-κB pathway in LPS-challenged HUVECs and septic mice.	Life Sci	3.647
14	LncRNA TUG1 regulates the balance of HuR and miR-29b-3p and inhibits intestinal epithelial cell apoptosis in a mouse model of ulcerative colitis.	Hum Cell	3.463
15	Circular RNA RBM33 contributes to cervical cancer progression via modulation of the miR-758-3p/PUM2 axis.	J Mol Histol	2.531
16	LncRNA BCYRN1-induced autophagy enhances asparaginase resistance in extranodal NK/T-cell lymphoma.	Theranostics	8.579
17	Oscillating lncRNA Platr4 regulates NLRP3 inflammasome to ameliorate nonalcoholic steatohepatitis in mice.	Theranostics	8.579
18	Long non-coding RNA MEG3 silencing and microRNA-214 restoration elevate osteoprotegerin expression to ameliorate osteoporosis by limiting TXNIP	J Cell Mol Med	4.486
19	PGM5-AS1 impairs miR-587-mediated GDF10 inhibition and abrogates progression of prostate cancer	J Transl Med	4.124
20	Circular RNA circLMO7 acts as a microRNA-30a-3p sponge to promote gastric cancer progression via the WNT2/β-catenin pathway	J Exp Clin Cancer Res	7.068
21	CircWHSC1 serves as an oncogene to promote hepatocellular carcinoma progression.	Eur J Clin Invest	3.481
22	Aerobic exercise improves VCI through circRIMS2/miR-186/BDNF-mediated neuronal apoptosis.	Mol Med	4.096
23	ZFPM2-AS1 facilitates cell proliferation and migration in cutaneous malignant melanoma through modulating miR-650/NOTCH1 signaling.	Dermatol Ther	2.327
24	Exosomal miR-186 derived from BMSCs promote osteogenesis through hippo signaling pathway in postmenopausal osteoporosis.	J Orthop Surg Res	1.777
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26	Extracellular vesicles derived from hypoxic glioma stem-like cells confer temozolomide resistance on glioblastoma by delivering miR-30b-3p.	Theranostics	8.579
27	Linc00887 suppresses tumorigenesis of cervical cancer through regulating the miR-454-3p/FRMD6-Hippo axis.	Cancer Cell Int	4.175
28	Circ-CCDC66 upregulates REXO1 expression to aggravate cervical cancer progression via restraining miR-452-5p.	Cancer Cell Int	4.175
29	Hypermethylation-mediated downregulation of lncRNA PVT1 promotes granulosa cell apoptosis in premature ovarian insufficiency via interacting with Foxo3a.	J Cell Physiol	5.546
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37	Circ-FOXM1 knockdown suppresses non-small cell lung cancer development by regulating the miR-149-5p/ATG5 axis.	Cell Cycle	3.699
38	Downregulation of lncRNA TSLNC8 promotes melanoma resistance to BRAF inhibitor PLX4720 through binding with PP1α to re-activate MAPK signaling.	J Cancer Res Clin Oncol	3.656
39	Linc01559 Served as a Potential Oncogene and Promoted Resistance of Hepatocellular Carcinoma to Oxaliplatin by Directly Sponging miR-6783-3p.	Anticancer Agents Med Chem	2.049
40	Knockdown of RMST Impedes Neuronal Apoptosis and Oxidative Stress in OGD/R-Induced Ischemic Stroke Via Depending on the miR-377/SEMA3A Signal Network.	Neurochem Res	3.038
41	CircRNA DNA methyltransferase 1 silence inhibits breast cancer development by regulating micoRNA-485-3p/zinc finger E-box binding homeobox 1 axis.	J Obstet Gynaecol Res	1.392
42	Diagnostic, prognostic, and therapeutic significance of long non-coding RNA MALAT1 in cancer.	Biochim Biophys Acta Rev Cancer	7.365
43	LncRNA NEAT1 regulates the proliferation and production of the inflammatory cytokines in rheumatoid arthritis fibroblast-like synoviocytes by targeting miR-204-5p.	Hum Cell	3.463
44	Silenced lncRNA H19 and up-regulated microRNA-129 accelerates viability and restrains apoptosis of PC12 cells induced by Aβ25-35 in a cellular model of Alzheimer's disease.	Cell Cycle	3.699
45	LncRNA HOTTIP leads to osteoarthritis progression via regulating miR-663a/ Fyn-related kinase axis.	BMC Musculoskelet Disord	1.879
46	Upregulation of amplified in breast cancer 1 contributes to pancreatic ductal adenocarcinoma progression and vulnerability to blockage of hedgehog activation.	Theranostics	8.579
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48	Astragaloside IV alleviates atherosclerosis through targeting circ_0000231/miR-135a-5p/CLIC4 axis in AS cell model in vitro.	Mol Cell Biochem	2.795
49	Circular RNA circ_0020123 Promotes Non-Small Cell Lung Cancer Progression Through miR-384/TRIM44 Axis.	Cancer Manag Res	2.886
50	Baicalin suppresses the progression of preeclampsia via regulating miR-155-5p/FOXO3 axis.	J Recept Signal Transduct Res	1.466
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52	LncRNA HOTTIP facilitates cell proliferation, invasion, and migration in osteosarcoma by interaction with PTBP1 to promote KHSRP level.	Cell Cycle	3.699
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65	LncRNA HOXA-AS2 promotes glioblastoma carcinogenesis by targeting miR-885-5p/RBBP4 axis.	Cancer Cell Int	4.175
66	Circ_0004771 Accelerates Cell Carcinogenic Phenotypes via Suppressing miR-1253-Mediated DDAH1 Inhibition in Breast Cancer.	Cancer Manag Res	2.886
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