依托咪酯通过调控LncRNA CDKN2B-AS1/miR-199a-5p表达抑制胶质瘤细胞增殖、迁移和侵袭

目的 探讨依托咪酯对胶质瘤细胞的增殖、迁移和侵袭的影响及其作用机制.方法 用不同浓度的依托咪酯处理胶质瘤细胞U251,采用四甲基偶氮唑蓝(MTT)检测依托咪酯对U251细胞增殖能力的影响;Transwell实验检测依托咪酯对U251细胞迁移及侵袭的影响;实时荧光定量-聚合酶链反应(qRT-PCR)检测胶质瘤组织中长链非...     

Clinical usefulness of gastric adenocarcinoma predictive long intergenic noncoding RNA in human malignancies: A meta-analysis

BackgroundGastric adenocarcinoma predictive long intergenic noncoding RNA (GAPLINC), a newly identified lincRNA, was reported to be aberrantly expressed in several kinds of cancers and played an important role in tumor progression. This study was performed to systematically estimate the prognostic role of GAPLINC expression in cancer patients.MethodsSeveral electronic databases, including PubMed, Embase, Web of Science, China National Knowledge Infrastructure (CNKI), and Wan Fang databases were searched for potential literature (updated to September 3, 2018). The pooled hazard ratios (HRs), odds ratios (ORs) and 95% confidence intervals (CIs) were calculated with a fixed effects model using Stata12.0 software.ResultsThe pooled results indicated that elevated GAPLINC was significantly related to shorter overall survival (OS) (HR = 1.66, 95%CI: 1.40–1.93, p < 0.001), which was further validated using The Cancer Genome Atlas (TCGA) dataset. Furthermore, high GAPLINC expression was correlated with higher tumor grade (OR = 1.91, 95%CI: 1.35–2.70, p < 0.001), positive lymph node metastasis (OR = 2.80, 95%CI: 1.69–4.64, p < 0.001), deeper infiltration depth (OR = 2.44, 95%CI: 1.43–4.17, p = 0.001) and advanced clinical stage (OR = 3.54, 95%CI: 2.13–5.88, p < 0.001).ConclusionsOur results suggest that elevated GAPLINC was associated with poor clinical outcomes and might serve as a promising prognostic biomarker in cancer survivors.     

非编码RNA调节心肌缺血再灌注损伤中自噬的作用及机制

背景:越来越多的研究表明自噬在心肌缺血再灌注损伤中发挥重要作用,适度的自噬有助于维持心脏的正常和代谢功能.非编码RNAs包括长链非编码RNA(lncRNAs)、环状RNA(circRNAs)和微小RNA(miRNAs),能够通过调控自噬参与维持心肌缺血再灌注损伤进程,从而维持心肌细胞稳态和保护心肌细胞.目的:对非编码R...     

LncRNA GAS5通过miR-21调控脊髓损伤后神经细胞凋亡的机制研究

目的 明确长链非编码RNA生长阻滞特异性转录因子5（LncRNA GAS5）与微小RNA-21（miR-21）的关系,阐明LncRNA GAS5调控miR-21参与脊髓损伤后神经细胞凋亡的机制。方法 建立脊髓损伤大鼠和氧糖剥夺复氧（OGD/R）处理的大鼠肾上腺嗜铬细胞瘤细胞（PC-12）模型;沉默和过表达GAS5,构建下调shGAS5表达的慢病毒,通过生物信息学分析预测LncRNA GAS5与miR-21存在结合位点等。通过双荧光素酶报告基因实验等探究GAS5与miR-21的结合位点;采用实时荧光定量聚合酶链式反应（RT-qPCR）检测miR-21、GAS5、同源性磷酸酶-张力蛋白（phosphatase and tensin homolog,PTEN）基因、半胱天冬氨酸蛋白酶3（Caspase 3）、B淋巴细胞瘤（Bcl）-2相关X基因（Bax）、Bcl-2和蛋白激酶B（AKT）的RNA表达水平;Western blot检测Cleaved caspase 3、Bax、Bcl-2、AKT和磷酸化AKT（p-AKT）的蛋白表达水平;TUNEL技术检测神经细胞凋亡程度。结果 大鼠脊髓损伤后脊髓中miR-21、Bcl-2及p-AKT表达水平降低（P＜0.05）,GAS5、PTEN、Bax及Cleaved caspase-3表达均升高（P＜0.05）。PC-12体外培养与OGD/R损伤后出现与大鼠脊髓损伤模型类似结果,且于OGD/R处理后2 h最显著。GAS5可通过碱基互补配对方式结合miR-21,进而调控下游PTEN信号通路。下调GAS5或过表达miR-21可抑制PC-12细胞凋亡（P＜0.05）。结论 GAS5通过结合miR-21,上调PTEN并抑制AKT磷酸化,进而促进脊髓损伤诱导的神经细胞凋亡。     

LncRNA ITGB2-AS1靶向miR-338-3p对肺癌细胞多西他赛耐药性的影响

目的 研究lncRNA ITGB2-AS1对肺癌细胞增殖、凋亡及多西他赛耐药性的影响及潜在的分子机制.方法 用不同浓度(6.25μg/L、12.5μg/L、25μg/L、50μg/L、100μg/L、200μg/L)多西他赛(DTX)处理肺癌A549和多西他赛耐药细胞A549/DTX细胞,CCK8法测定DTX对A549...     

K562细胞DHRS4 L1基因新选择性剪接亚型的克隆及二级结构预测

目的： K562细胞DHRS4L1[ dehydrogenase／reductase （ SDR family） member 4 like 1]的表达及其二级结构分析。方法以K562细胞cDNA为模板, PCR扩增DHRS4[ dehydrogenase／reductase （ SDR fami-ly） member 4]基因簇Ea2转录本。将PCR产物A-T克隆至pGEMT-Easy质粒,对质粒进行DNA Sanger测序。将测序所得序列用NCBI ORF finder分析是否存在编码区,用Clustal Omega分析RNA序列系统树,用RNAfold web server分析RNA最小自由能二级结构。结果用RT-PCR和Sanger测序方法发现, K562表达DHRS4L1 Ea2转录本,未检测到其表达DHRS4L2[dehydrogenase／reductase （SDR family） member 4 like 2] Ea1转录本。 K562表达的DHRS4L1 Ea2至少有8种选择性剪接亚型（ KU058702、 KU058703、 KU058704、KU058705、 KU058706、 KU058707、 KU058708、 KU058709）,其中6种为首次发现。 KU058702、 KU058703、KU058704、 KU058705和KU058707第二外显子受到多种形式的选择性剪接,恰好仅影响第2臂的二级结构,不影响其他臂的二级结构,提示这些不同亚型的二级结构有一定相似性,第二外显子所对应的二级结构臂可能在区分不同 DHRS4L1亚型功能中发挥关键作用。结论研究发现 K562细胞至少表达8种DHRS4L1选择性剪接亚型,为长链非编码RNA。其二级结构分析为后续研究DHRS4L1在白血病细胞中的潜在功能奠定基础。     

长链非编码RNA OPA相互作用蛋白5反向转录序列1靶向调控微RNA-128-3p对结直肠癌细胞增殖、侵袭和转移的影响

目的 探究长链非编码RNA OPA相互作用蛋白5反向转录序列1(lncRNA OIP5-AS1)在结直肠癌中的表达情况及靶向调控微RNA-128-3p(miR-128-3p)对结直肠癌细胞增殖、侵袭和转移的影响。方法 采用实时荧光定量PCR(qPCR)定量分析2017年1月至2018年12月在郑州大学人民医院住院并进行手术治疗的38例结直肠癌病人癌组织及相应癌旁组织、结直肠癌细胞系（SW480、SW620、HT-29和LoVo）及人正常结直肠黏膜细胞FHC中lncRNA OIP5-AS1和miR-128-3p的表达。将SW620细胞设为si-OIP5-AS1组、si-NC组、miR-128-3p mimic组、mimic-NC组、miR-128-3p inhibitor+si-OIP5-AS1组和inhibitor-NC+siOIP5-AS1组,采用细胞计数试剂盒（CCK-8）实验和克隆形成实验检测细胞增殖能力,采用划痕实验和transwell实验检测细胞侵袭与迁移能力,采用蛋白质印迹法检测E2F1、细胞周期蛋白D1(Cyclin D1)、波形蛋白（Vimentin）、N-钙黏蛋白（N...     

Modulatory Potential of LncRNA Zfas1 for Inflammation and Neuronal Apoptosis in Temporal Lobe Epilepsy

PurposeThis study aimed to elucidate whether lncRNA ZFAS1 is involved in neuronal apoptosis and inflammation in temporal lobe epilepsy (TLE).Materials and MethodsNinety-six TLE patients were recruited, and their peripheral venous blood was gathered to determine Zfas1 expression with polymerase chain reaction. Neurons were separated from hippocampal tissue of newborn SD rats, and si-Zfas1 or pcDNA3.1-Zfas1 was transfected into the neurons. Inflammatory cytokines released by neurons were determined, and neuronal activities were evaluated through MTT assay, colony formation assay, and flow cytometry.ResultsSerum levels of Zfas1 were higher in TLE patients than in healthy controls (p<0.05). Furthermore, Zfas1 expression in neurons was raised by pcDNA3.1-Zfas1 and declined after silencing of Zfas1 (p<0.05). Transfection of pcDNA-Zfas1 weakened the viability and proliferation of neurons and increased neuronal apoptosis (p<0.05). Meanwhile, pcDNA3.1-Zfas1 transfection promoted lipopolysaccharide-induced release of cytokines, including tumor necrosis factor-α, interleukin (IL)-1, IL-6, and intercellular adhesion molecule-1 (p<0.05), and boosted NF-κB activation by elevating the expression of NF-κB p65, pIκBα, and IKKβ in neurons (p<0.05).ConclusionOur results indicated that lncRNA ZFAS1 exacerbates epilepsy development by promoting neuronal apoptosis and inflammation, implying ZFAS1 as a promising treatment target for epilepsy.     

Expression of lncRNA NEAT1 in peripheral blood mononuclear cells of patients with systemic lupus erythematosus and its correlation with Th1/Th2 balance

Objective: This study explored and analyzed the expression of LncRNA NEAT1 in peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE) and its correlation with Th1/Th2 balance. Methods: We chose 97 SLE patients admitted in our hospital from Jun. 2016 to Feb. 2019 as SLE group, and randomly selected 50 healthy volunteers that underwent physical examination in our hospital during the same period as control group. We detected the expression of LncRNA NEAT1 in PBMCs of the two groups of subjects by qRT-PCR, the degree of Th1 and Th2 cells in both groups by flow cytometry, and the expression of TFN-γ and IL-4 in both groups by ELISA. Results: The relative expression of LncRNA NEAT1 in PBMCs of SLE group was higher than that of control group (P<0.05). The proportion of Th1 and the ratio of Th1/Th2 cells in PBMCs were markedly lower in the SLE group than the control group (P<0.05), while the proportion of Th2 was higher in the SLE group than the control group (P<0.05). IFN-γ level in SLE group was much lower than the control group (P<0.05), while IL-4 level was evidently higher in the SLE group than in controls (P<0.05). The expression of LncRNA NEAT1 in PBMCs of SLE group was notably negatively correlated with Th1 proportion and Th1/Th2 ratio (P<0.05), while positively correlated with Th2 proportion (P<0.05). Conclusion: LncRNA NEAT1 in PBMCs of SLE patients is abnormally highly expressed, and this expression is negatively correlated with Th1/Th2 balance. These two factors may interact and jointly affect the occurrence and progression of SLE.