BACKGROUND & AIMS: :Epithelial ovarian cancer (EOC) is the leading cause of death among gynecologic malignancies. Despite great efforts to improve early detection and optimize chemotherapeutic regimens, the 5-year survival rate is only 30% for patients presenting with late-stage ovarian cancer. The high mortality of this disease is due to late diagnosis in over 70% of ovarian cancer cases. A class of small noncoding RNAs, or microRNAs, was found to regulate gene expression at the post-transcriptional level. Some, but not all, of the data indicated that the miR-200 family was dysregulated in a variety of malignancies. In this study, we demonstrated that miR-200a and E-cadherin were significantly upregulated in EOC compared to benign epithelial ovarian cysts and normal ovarian tissues. However, further stratification of the subject indicated that the expression levels of miR-200a were significantly downregulated in late-stage (FIGO III+V) and grade 3 groups compared with early stage (FIGO I+II) and grade 1 to 2 groups. Similarly, relatively low levels of miR-200a were observed in the lymph compared to the node-negative group. E-cadherin expression was found to be absent in normal ovarian tissue and was frequently expressed in benign epithelial ovarian cysts, with absence or low levels observed in late-stage ovarian cancers. There was a significantly positive correlation between miR-200a and E-cadherin in EOC. The biphasic expression pattern suggested that miR-200a levels may serve as novel biomarkers for the early detection of EOC, and miR-200a and E-cadherin are candidate targets for the development of new treatment modalities against ovarian cancer.

背景与目标： : 上皮性卵巢癌 (EOC) 是妇科恶性肿瘤死亡的主要原因。尽管为改善早期检测和优化化疗方案做出了巨大努力，但对于晚期卵巢癌患者而言，5年生存率仅30%。这种疾病的高死亡率是由于超过70% 例卵巢癌病例的晚期诊断。发现一类小的非编码rna或microrna在转录后水平上调节基因表达。一些 (但不是全部) 数据表明，miR-200家族在各种恶性肿瘤中失调。在这项研究中，我们证明了与良性上皮性卵巢囊肿和正常卵巢组织相比，EOC中的miR-200a和E-cadherin显着上调。然而，对受试者的进一步分层表明，与早期 (FIGO I II) 和1至2级组相比，晚期 (FIGO III V) 和3级组的miR-200a表达水平显着下调。同样，与淋巴结阴性组相比，在淋巴中观察到相对较低的miR-200a水平。发现E-cadherin表达在正常卵巢组织中不存在，并且在良性上皮性卵巢囊肿中经常表达，在晚期卵巢癌中观察到不存在或低水平。EOC中miR-200a与E-cadherin之间存在显着正相关。双相表达模式表明，miR-200a水平可能是早期检测EOC的新型生物标志物，miR-200a和E-钙粘蛋白是开发针对卵巢癌的新治疗方式的候选靶标。

BACKGROUND & AIMS: :miRNAs are important regulators of cellular senescence yet the extent of their involvement remains to be investigated. We sought to identify miRNAs that are involved in cytokine-induced premature senescence (CIPS) in endothelial cells. CIPS was established in young human pulmonary microvascular endothelial cells (HMVEC-Ls) following treatment with a sublethal dose (20ng/ml) of tumor necrosis factor alpha (TNF-α) for 15days. In parallel, HMVEC-Ls were grown and routinely passaged until the onset of replicative senescence (RS). Differential expression analysis following miRNA microarray profiling revealed an overlapped of eight deregulated miRNAs in both the miRNA profiles of RS and TNF-α-induced premature senescence cells. Amongst the deregulated miRNAs were members of the miR 17-92 cluster which are known regulators of angiogenesis. The role of hsa-miR-20b in TNF-α-induced premature senescence, a paralog member of the miR 17-92 cluster, was further investigated. Biotin-labeled hsa-miR-20b captured the enriched transcripts of retinoblastoma-like 1 (RBL1), indicating that RBL1 is a target of hsa-miR-20b. Knockdown of hsa-miR-20b attenuated premature senescence in the TNF-α-treated HMVEC-Ls as evidenced by increased cell proliferation, increased RBL1 mRNA expression level but decreased protein expression of p16INK4a, a cellular senescence marker. These findings provide an early insight into the role of hsa-miR-20b in endothelial senescence.

背景与目标： : mirna是细胞衰老的重要调节剂，但其参与程度仍有待研究。我们试图鉴定参与细胞因子诱导的内皮细胞过早衰老 (CIPS) 的miRNAs。用亚致死剂量 (20ng/ml) 的肿瘤坏死因子 α (TNF-α) 治疗15天后，在年轻的人肺微血管内皮细胞 (HMVEC-Ls) 中建立了CIPS。同时，HMVEC-Ls生长并常规传代，直到复制性衰老 (RS) 开始。miRNA微阵列分析后的差异表达分析显示，在RS和TNF-α 诱导的早衰细胞的miRNA谱中，八个解除调节的miRNA重叠。在解除管制的mirna中，miR 17-92簇的成员是已知的血管生成调节剂。进一步研究了hsa-miR-20b在TNF-α 诱导的过早衰老 (miR 17-92簇的旁系成员) 中的作用。生物素标记的hsa-miR-20b捕获了视网膜母细胞瘤样1 (RBL1) 的富集转录本，表明RBL1是hsa-miR-20b的靶标。hsa-miR-20b的敲除减弱了TNF-α 处理的HMVEC-Ls中的过早衰老，这可以通过增加细胞增殖，增加RBL1 mRNA表达水平但降低细胞衰老标志物p16INK4a的蛋白表达来证明。这些发现提供了hsa-miR-20b在内皮衰老中的作用的早期见解。

BACKGROUND & AIMS: OBJECTIVES:MicroRNAs (miRNAs) are endogenous small RNAs of 21-25 nucleotides that can pair with sites in 3' untranslated regions in mRNAs of protein-coding genes to downregulate their expression. Recently, miR-499 and other miRNAs released in circulating blood have been reported as promising biomarkers for acute myocardial infarction (AMI). In the present study, we developed a novel reverse-transcription real-time PCR assay for human miR-499 quantification. DESIGN AND METHODS:miR-499 was reverse-transcribed with a 3' portion-specific primer into cDNAs. The cDNAs were further extended with overlap PCR. The extended cDNAs were determined by quantitative, real-time PCR. Synthetic miR-499 was put into the RT reaction over an optimal range to generate standard curves for absolute quantification of miR-499. RESULTS:In the presence of 0.0001 amol/μL to 1.0×10⁶ amol/μL of synthetic miR-499, we observed a linear correlation (R²=0.999) between the logarithm of the amount of input RNA and the CT value. The miR-499 was reliably measured at a detection limit of 0.0001 amol/μL. The miR-499 measurements in spiked plasma samples indicated excellent correlation between the novel qRT PCR and classic stem loop qRT PCR. The qRT PCR analysis demonstrated that miR-499 was detected with higher levels in plasma from the patient with AMI in acute phase (AMI) compared with those from the control groups (P<0.001). CONCLUSIONS:We developed a novel reverse-transcription real-time PCR assay for human miR-499 quantification. The good reproducibility and wide linearity range may permit more use of it in the quantification of other human miRNAs in future.

背景与目标：

BACKGROUND & AIMS: :Patients with secondary acute myeloid leukemia (sAML) arising from myelodysplastic syndromes have a poor prognosis marked by an increased resistance to chemotherapy. An urgent need exists for adjuvant treatments that can enhance or replace current therapeutic options. Here we show the potential of LB100, a small-molecule protein phosphatase 2 A (PP2A) inhibitor, as a monotherapy and chemosensitizing agent for sAML using an in-vitro and in-vivo approach. We demonstrate that LB100 decreases cell viability through caspase activation and G2/M cell-cycle arrest. LB100 enhances daunorubicin (DNR) cytotoxicity resulting in decreased xenograft volumes and improved overall survival. LB100 profoundly upregulates miR-181b-1, which we show directly binds to the 3' untranslated region of Bcl-2 mRNA leading to its translational inhibition. MiR-181b-1 ectopic overexpression further diminishes Bcl-2 expression leading to suppression of sAML cell growth, and enhancement of DNR cytotoxicity. Our research highlights the therapeutic potential of LB100, and provides new insights into the mechanism of LB100 chemosensitization.

背景与目标： : 由骨髓增生异常综合征引起的继发性急性髓细胞性白血病 (sAML) 患者的预后较差，其对化疗的抵抗力增加。迫切需要可以增强或替代当前治疗选择的辅助治疗。在这里，我们使用体外和体内方法显示了小分子蛋白磷酸酶2 a (PP2A) 抑制剂LB100作为sAML的单一疗法和化学增敏剂的潜力。我们证明LB100通过caspase激活和G2/M细胞周期阻滞降低细胞活力。LB100增强柔红霉素 (DNR) 的细胞毒性，从而减少异种移植体积并改善总生存率。LB100深刻上调miR-181b-1，我们显示其与Bcl-2 mRNA的3' 非翻译区直接结合，导致其翻译抑制。MiR-181b-1异位过度表达进一步减少Bcl-2表达，导致sAML细胞生长的抑制和DNR细胞毒性的增强。我们的研究突出了LB100的治疗潜力，并为LB100的化学增敏机制提供了新的见解。

BACKGROUND & AIMS: :Human cytomegalovirus (HCMV) has been reported to be linked to vascular disease through the induction of neovessel formation. We have previously reported that microRNA (miR)-217 and miR-199a-5p enhance endothelial angiogenesis via inhibition of sirtuin 1 (SIRT1) in HCMV-infected human umbilical vein endothelial cells (HUVECs). Here, we found that miR-138 also suppressed the expression of the SIRT1 protein and stimulated phosphorylation of signal transducer and activator of transcription 3 (p-STAT3). Moreover, the regulation of p-STAT3 expression mediated by SIRT1 was found to promote HCMV-induced angiogenesis. These findings revealed that miR-138 might promote angiogenesis of HCMV-infected HUVECs by activating the SIRT1-mediated p-STAT3 pathway, and this could provide novel insights into HCMV-induced angiogenesis.

背景与目标： : 据报道，人类巨细胞病毒 (HCMV) 通过诱导新血管形成与血管疾病有关。我们先前已经报道了microRNA (miR)-217和miR-199a-5p通过抑制HCMV感染的人脐静脉内皮细胞 (huvec) 中的sirtuin 1 (SIRT1) 来增强内皮血管生成。在这里，我们发现miR-138还抑制了SIRT1蛋白的表达并刺激了信号转导子和转录激活子3的磷酸化 (p-STAT3)。此外，发现由SIRT1介导的p-STAT3表达调节可促进HCMV诱导的血管生成。这些发现表明，miR-138可能通过激活SIRT1-mediated p-STAT3途径促进HCMV感染的HUVECs的血管生成，这可能为HCMV诱导的血管生成提供新的见解。

BACKGROUND & AIMS: :The expression pattern and role of circular RNAs (circRNAs) in the pathogenesis of gastric cancer (GC) and their underlying mechanisms remain unresolved. In this study, we identified differentially expressed circRNAs by a circRNA microarray and verified the results by quantitative reverse transcription-polymerase chain reaction using 117 clinical samples. Cell Counting Kit-8, wound healing, Transwell, and tumorsphere formation assays were conducted to assess the effects of circ-CEP85L on cell proliferation and invasion in vitro. Mouse intraperitoneal injection models were used to assess the functions of circ-CEP85L in vivo. Luciferase reporter assays, fluorescence in situ hybridization, and rescue experiments were performed to elucidate the underlying mechanism of circ-CEP85L. We found that circ-CEP85L, which has not been studied in GC, was significantly downregulated in GC tissues and that decreased circ-CEP85L expression correlated significantly with a worse prognosis. The knockdown of circ-CEP85L promoted the proliferation and invasion of GC cells, which was reversed by overexpression of circ-CEP85L. Furthermore, inhibition of circ-CEP85L promoted tumor growth in vivo. Mechanistically, circ-CEP85L was confirmed to be a direct target of miR-942-5p. In addition, rescue experiments indicated that circ-CEP85L is able to inhibit the proliferation and invasion of GC cells by sponging miR-942-5p. Finally, western blot assays verified that the downregulation of miR-942-5p efficiently reversed the inhibition of NFKBIA induced by circ-CEP85L overexpression. Therefore, we conclude that circ-CEP85L promotes NFKBIA expression by acting as a sponge of miR-942-5p; thus, inhibiting GC proliferation and invasion. circ-CEP85L is a potential target in the treatment of GC.

背景与目标： : 环状rna (circRNAs) 在胃癌 (GC) 发病中的表达模式和作用及其潜在机制仍未解决。在这项研究中，我们通过circRNA微阵列鉴定了差异表达的circRNA，并使用117临床样品通过定量逆转录-聚合酶链反应验证了结果。进行细胞计数试剂盒-8，伤口愈合，Transwell和肿瘤球形成试验，以评估circ-CEP85L对体外细胞增殖和侵袭的影响。小鼠腹腔注射模型用于评估体内circ-CEP85L的功能。进行了荧光素酶报告基因测定，荧光原位杂交和抢救实验，以阐明circ-CEP85L的潜在机制。我们发现，尚未在GC中进行研究的circ-CEP85L在GC组织中被显着下调，并且circ-CEP85L表达的降低与预后差显着相关。circ-CEP85L的敲除促进了GC细胞的增殖和侵袭，而circ-CEP85L的过表达则使其逆转。此外，抑制circ-CEP85L促进体内肿瘤生长。从机械上讲，circ-CEP85L被确认为miR-942-5p的直接目标。此外，拯救实验表明，circ-CEP85L能够通过海绵miR-942-5p抑制GC细胞的增殖和侵袭。最后，western blot测定证实了miR-942-5p的下调有效地逆转了circ-CEP85L过表达诱导的NFKBIA的抑制作用。因此，我们得出结论，circ-CEP85L通过充当miR-942-5p海绵来促进NFKBIA表达; 因此，抑制GC增殖和侵袭。circ-CEP85L是GC医治的一个潜在靶点。

BACKGROUND & AIMS: :Mirtrons are non-canonical miRNAs arising by splicing and debranching from short introns. A plethora of introns have been inferred by computational analyses as potential mirtrons. Yet, few have been experimentally validated and their functions, particularly in relation to their host genes, remain poorly understood. Here, we found that Drosophila larvae lacking either the mirtron miR-1010 or its binding site in the nicotinic acetylcholine receptor β2 (nAcRβ2) 3'UTR fail to grow properly and pupariate. Increase of cortical nAcRβ2 mediated by neural activity elevates the level of intracellular Ca2+, which in turn activates CaMKII and, further downstream, the transcription factor Adf-1. We show that miR-1010 downregulates nAcRβ2. We reveal that Adf-1 initiates the expression of SKIP, the host gene of miR-1010. Preventing synaptic potentials from overshooting their optimal range requires both SKIP to temper synaptic potentials (incoherent feedforward loop) and miR-1010 to reduce nAcRβ2 mRNA levels (negative feedback loop). Our results demonstrate how a mirtron, in coordination with its host gene, contributes to maintaining appropriate receptor levels, which in turn may play a role in maintaining homeostasis.

背景与目标： : Mirtrons是由短内含子剪接和脱支产生的非规范mirna。通过计算分析，已经推断出过多的内含子是潜在的mirtrons。然而，很少有人得到实验验证，它们的功能，特别是与宿主基因有关的功能，仍然知之甚少。在这里，我们发现果蝇幼虫在烟碱乙酰胆碱受体 β2 (nAcRβ2) 3'UTR中缺乏mirtron miR-1010或其结合位点，无法正常生长和腐烂。由神经活动介导的皮质nAcRβ2的增加会提高细胞内Ca2的水平，进而激活CaMKII，并在更下游激活转录因子Adf-1。我们显示miR-1010下调nacrβ2。我们揭示了Adf-1启动miR-1010宿主基因SKIP的表达。防止突触电位超出其最佳范围需要跳过以调节突触电位 (非相干前馈环) 和miR-1010以降低nacrβ2mrna水平 (负反馈环)。我们的结果证明了mirtron如何与其宿主基因协调，有助于维持适当的受体水平，进而可能在维持体内平衡中发挥作用。

BACKGROUND & AIMS: INTRODUCTION:MicroRNAs (miRNAs) are small non-coding single-stranded RNA molecules that regulate gene expression at the post-transcriptional level. In the pathogenesis of chronic lymphocytic leukemia (CLL), miR-15a and miR-16-1 play an important role. These miRNAs are located on chromosome 13 in the 13q14.3 region, which is deleted in more than 55% of CLL patients. This aberration affects expression of miRNAs. OBJECTIVES:The study aimed at performing a molecular genetic analysis of miR-15a and miR-16-1 expression in a group of 39 patients diagnosed with CLL and determining the association between the expression of the two miRNAs and types of deletions in the 13q14 region. METHODS:We used fluorescence in situ hybridiziation (FISH) for determination of mono- or biallelic deletion 13q and quantitative polymerase chain reaction (Q-RT-PCR) to revealed expression miR-15a and miR-16-1 in 39 patients suffering from CLL. RESULTS:The analysis comprised 19 patients with monoallelic 13q14 deletion, 3 patients with biallelic deletion, 9 patients with both monoallelic and biallelic deletions, and 8 patients without 13q14 deletion serving as controls. The results showed different levels of miRNA expression in individual patients. Significantly higher normalized levels of miR-15a expression were found in the control group and patients with monoallelic 13q14 expression compared with patients with biallelic deletion. There was a significantly decreased expression of both miRNAs in patients with biallelic deletion of the 13q14 region but only when deletions were present in 77% or more of cells, as detected by fluorescent in situ hybridization (FISH).

背景与目标：

BACKGROUND & AIMS: :Endothelial progenitor cell (EPC) recruitment and angiogenesis play crucial roles in aneurysm neck endothelialization, but the mechanisms of EPC recruitment and angiogenesis are still unclear. Recent studies have shown that long noncoding RNAs (lncRNAs) can regulate the function and differentiation of cells in various ways. LncRNA TUG1 is involved in liver cancer and glioma-mediated angiogenesis. The aim of this study was to investigate the role of lncRNA TUG1 in regulating EPC migration and differentiation. Overexpression and knockdown of lncRNA TUG1 with lentivirus, scratch assays, Transwell assays and tube formation assays using EPCs isolated from rat bone marrow showed that lncRNA TUG1 overexpression promoted EPC migration, invasion and differentiation. Moreover, ELISAs showed that lncRNA TUG1 overexpression increased VEGF expression. Bioinformatics prediction, luciferase assays, Western blots and RIP assays indicated that lncRNA TUG1 functions as a ceRNA (competing endogenous RNA) for miR-6321 and that miR-6321 inhibits EPC migration and differentiation through its target, ATF2. As a potential therapeutic target, lncRNA TUG1 may play a vital role in the pathogenesis of aneurysms.

背景与目标： 内皮祖细胞 (EPC) 募集和血管生成在动脉瘤颈部内皮化中起着至关重要的作用，但EPC募集和血管生成的机制仍不清楚。最近的研究表明，长链非编码rna (lncRNAs) 可以通过多种方式调节细胞的功能和分化。LncRNA TUG1参与肝癌和神经胶质瘤介导的血管生成。这项研究的目的是研究lncRNA TUG1在调节EPC迁移和分化中的作用。使用从大鼠骨髓分离的EPC对lncRNA TUG1的过表达和敲除，划痕试验，tranwell试验和管形成试验表明，lncRNA TUG1的过表达促进了EPC的迁移，侵袭和分化。此外，ELISAs显示lncRNA TUG1过表达增加了VEGF的表达。生物信息学预测，荧光素酶测定，Western blots和RIP测定表明lncRNA TUG1充当miR-6321的ceRNA (竞争性内源性RNA)，并且miR-6321通过其靶标atf2抑制EPC的迁移和分化。作为潜在的治疗靶点，lncRNA TUG1可能在动脉瘤的发病机制中起着至关重要的作用。

BACKGROUND & AIMS: BACKGROUND:Due to the acquired drug resistance, the potency of cisplatin-based chemotherapy is limited in lung cancer, which is a big obstacle in clinical treatment of lung cancer. Abundant evidence has revealed that circular RNAs (circRNAs) exerted facilitating or suppressive function on the tumorigenesis of multiple cancers. The oncogenic role of circ-ABCB10 in breast cancer and clear cell renal cell carcinoma has been validated in recent researches. However, the regulatory mechanism of circ-ABCB10 and its relation to cellular sensitivity to cisplatin in lung cancer is poorly understood. METHODS:The expression and characteristic of circ-ABCB10 were analyzed by RT-qPCR and nucleic acid electrophoresis. CCK-8, colony formation, TUNEL and transwell assays were applied to probe the role of FOXD3-AS1 in lung cancer. The interactions of miR-556-3p with circ-ABCB10 and AK4 were testified by luciferase reporter and RIP assays. RESULTS:Circ-ABCB10 was markedly upregulated and featured with loop structure in lung cancer. Circ-ABCB10 depletion suppresses lung cancer progression and sensitizes lung cancer cells to cisplatin. Molecular mechanism assays manifested that circ-ABCB10 bound with miR-556-3p and negatively modulated miR-556-3p expression. Additionally, AK4 was testified to be the downstream target of miR-556-3p. More importantly, rescue assays clarified that upregulation of AK4 could reverse the cisplatin-sensitizing and tumor-suppressing effect of circ-ABCB10 knockdown on lung cancer cells. CONCLUSIONS:Circ-ABCB10 knockdown enhances sensitivity of lung cancer cells to cisplatin by targeting miR-556-3p/AK4 axis.

背景与目标：

BACKGROUND & AIMS: :Neutrophils are implicated in the pathogenesis of atherosclerosis but are seldom detected in atherosclerotic plaques. We investigated whether neutrophil-derived microvesicles may influence arterial pathophysiology. Here we report that levels of circulating neutrophil microvesicles are enhanced by exposure to a high fat diet, a known risk factor for atherosclerosis. Neutrophil microvesicles accumulate at disease-prone regions of arteries exposed to disturbed flow patterns, and promote vascular inflammation and atherosclerosis in a murine model. Using cultured endothelial cells exposed to disturbed flow, we demonstrate that neutrophil microvesicles promote inflammatory gene expression by delivering miR-155, enhancing NF-κB activation. Similarly, neutrophil microvesicles increase miR-155 and enhance NF-κB at disease-prone sites of disturbed flow in vivo. Enhancement of atherosclerotic plaque formation and increase in macrophage content by neutrophil microvesicles is dependent on miR-155. We conclude that neutrophils contribute to vascular inflammation and atherogenesis through delivery of microvesicles carrying miR-155 to disease-prone regions.

背景与目标： 中性粒细胞与动脉粥样硬化的发病机制有关，但很少在动脉粥样硬化斑块中检测到。我们调查了中性粒细胞来源的微泡是否会影响动脉病理生理学。在这里，我们报告了通过暴露于高脂饮食 (一种已知的动脉粥样硬化危险因素) 来提高循环中性粒细胞微泡的水平。中性粒细胞微泡在暴露于紊乱血流模式的动脉易患病区域积聚，并在鼠模型中促进血管炎症和动脉粥样硬化。使用暴露于扰动流的培养的内皮细胞，我们证明中性粒细胞微泡通过递送miR-155促进炎症基因表达，增强NF-κ b活化。同样，中性粒细胞微泡增加miR-155，并在体内流动紊乱的易发部位增强NF-κ b。中性粒细胞微泡对动脉粥样硬化斑块形成的增强和巨噬细胞含量的增加取决于miR-155。我们得出的结论是，中性粒细胞通过将携带miR-155的微泡递送到易患疾病的区域而导致血管炎症和动脉粥样硬化。

BACKGROUND & AIMS: :Acute myocardial infarction (AMI) results from long-term diminished blood supply diminishment (ischemia) to the heart, and the main reason for ischemia is hypoxia. BCL2 interaction protein 3 (BNIP3) can be upregulated by hypoxia and participates in the mediation of hypoxia-activated apoptosis in cardiac myocyte death. The purpose of this study was to interrogate the mechanism of BNIP3 in hypoxia-activated cardiac myocyte injury. Cell viability and apoptosis were evaluated by Cell counting kit 8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), TdT-mediated dUTP Nick-End Labeling (TUNEL), and caspase-3 activity assays. Molecular interactions were assessed by RNA immunoprecipitation (RIP) and pull-down assays. Gene levels were assessed via quantitative real-time PCR and western blot. BNIP3 expression was upregulated by hypoxia in H9c2 cells. We found that circ-BNIP3 (hsa_circ_0005972), whose annotated gene was BNIP3, was induced by hypoxia and positively regulated BNIP3 expression. Knockdown of BNIP3 or circ-BNIP3 reversed the effect of hypoxia in attenuating H9c2 cell viability and inducing apoptosis. circ-BNIP3 sponged miRNA-27a-3p (miR-27a-3p) to upregulate BNIP3 expression. Moreover, eukaryotic translation initiation factor 4A3 (EIF4A3) bound with the upstream region of the circ-BNIP3 mRNA transcript and induced circ-BNIP3 expression in H9c2 cells. EIF4A3-induced circ-BNIP3 aggravated hypoxia-caused injury of H9c2 cells through targeting miR-27a-3p/BNIP3 pathway, indicating circ-BNIP3 as a new target for relieving hypoxia-induced injury of cardiac myocytes.

背景与目标： : 急性心肌梗死 (AMI) 是由于心脏的长期血液供应减少 (缺血) 所致，而缺血的主要原因是缺氧。BCL2相互作用蛋白3 (BNIP3) 可被缺氧上调，并参与缺氧激活的心肌细胞凋亡的介导。目的探讨BNIP3在缺氧激活心肌细胞损伤中的作用机制。通过细胞计数试剂盒8 (CCK-8)，5-乙炔基-2 '-脱氧尿苷 (EdU)，TdT介导的dUTP缺口末端标记 (TUNEL) 和caspase-3活性测定来评估细胞活力和凋亡。通过RNA免疫沉淀 (RIP) 和下拉测定法评估分子相互作用。通过定量实时PCR和western blot评估基因水平。缺氧在H9c2细胞中BNIP3表达上调。我们发现低氧诱导了BNIP3基因的circ-BNIP3 (hsa_circ_0005972)，并正向调节了BNIP3的表达。敲除BNIP3或circ-BNIP3可逆转缺氧在减弱H9c2细胞活力和诱导凋亡中的作用。circ-BNIP3海绵miRNA-27a-3p (miR-27a-3p) 上调BNIP3表达。此外，真核翻译起始因子4A3 (EIF4A3) 与circ-BNIP3 mRNA转录物的上游区域结合，并在H9c2细胞中诱导circ-BNIP3表达。EIF4A3-induced circ-BNIP3通过靶向miR-27a-3p/BNIP3通路加重H9c2细胞缺氧损伤，提示circ-BNIP3是缓解缺氧诱导的心肌细胞损伤的新靶点。

BACKGROUND & AIMS: :Retinopathy of prematurity is a major cause of childhood blindness worldwide. Hence, exploring the proper treatment methods is a must in tacking this disease. qRT-PCR and western blot were used to detect the expression of genes and proteins, respectively. The proliferation of human retinal vascular endothelial cells (HRECs) was ensured by MTT assay. The luciferase activity was measured through luciferase assay. The inverted phase-contrast light microscope was used to observe the formation of a vascular tube. In the present study, our data demonstrated that circPDE4B was downregulated, while hypoxia-inducible factor-1α (HIF-1α) and VEGFA were upregulated in the retinopathy of prematurity model in vitro and in vivo. CircPDE4B increasing remarkably inhibited the expression of HIF-1α and VEGFA in hypoxia-induced HRECs and subsequent repressed cell proliferation and pathological angiogenesis. We further found that miR-181c suppressed the expression of von Hippel-Lindau (VHL), while circPDE4B could promote VHL expression via binding to miR-181c. Finally, our results revealed that circPDE4B inhibited the expression of VEGFA and pathological angiogenesis via facilitating VHL-mediated ubiquitin degradation of HIF-1α. In conclusion, circPDE4B suppressed the expression of VEGFA and pathological angiogenesis via promoting VHL-mediated ubiquitin degradation of HIF-1α through binding to miR-181c. Our study indicated that circPDE4B might be an effective therapeutic target of retinopathy of prematurity.

背景与目标： : 早产儿视网膜病变是全球儿童失明的主要原因。因此，探索正确的治疗方法是治疗这种疾病的必要条件。分别使用qRT-PCR和western blot检测基因和蛋白质的表达。通过MTT法确保人视网膜血管内皮细胞 (HRECs) 的增殖。荧光素酶活性通过荧光素酶法测定。使用倒置相差光学显微镜观察血管管的形成。在本研究中，我们的数据表明，在体外和体内的早产儿视网膜病变模型中，circPDE4B被下调，而缺氧诱导因子-1α (HIF-1α) 和VEGFA被上调。CircPDE4B的增加显着抑制了缺氧诱导的hrec中HIF-1α 和VEGFA的表达，随后抑制了细胞增殖和病理性血管生成。我们进一步发现miR-181c抑制了von Hippel-Lindau (VHL) 的表达，而circPDE4B可以通过与miR-181c结合来促进VHL的表达。最后，我们的结果表明，circPDE4B通过促进VHL介导的HIF-1α 的泛素降解来抑制VEGFA的表达和病理性血管生成。总之，circPDE4B通过与miR-181c结合促进VHL介导的HIF-1α 的泛素降解来抑制VEGFA的表达和病理性血管生成。我们的研究表明，circPDE4B可能是早产儿视网膜病的有效治疗靶标。

BACKGROUND & AIMS: :MicroRNA (miR)-802 has been discovered to be involved in the occurrence and development of numerous types of tumor; however, studies into the role of miR‑802 in cervical cancer are limited. Therefore, the present study aimed to investigate the regulatory effects of miR‑802 in cervical cancer cells. miR‑802 expression levels in cervical cancer tissue and cells were analyzed using reverse transcription‑quantitative (RT‑q)PCR, a dual‑reporter luciferase activity assay was used to identify the direct target gene of miR‑802, and RT‑qPCR and western blotting were performed to determine the relationship between miR‑802 and basic transcription factor 3 (BTF3). Cell viability, and migration and invasion were analyzed using Cell Counting Kit‑8 and Transwell assays, respectively. Finally, the expression levels of metastasis‑associated proteins, N‑cadherin and E‑cadherin, were determined using RT‑qPCR and western blotting. Decreased expression levels of miR‑802 were found in cervical cancer tissues and cells, and the overexpression of miR‑802 inhibited cell viability, migration and invasion. Moreover, miR‑802 was discovered to directly target BTF3 to inhibit its expression. Notably, the overexpression miR‑802 markedly reversed the promotive effect of BTF3 on cell viability, in addition to the migratory and invasive abilities of the cells. Simultaneously, the overexpression of miR‑802 significantly suppressed epithelial‑mesenchymal transition, and the expression levels of matrix metallopeptidase (MMP)2 and MMP9 in cells through regulating BTF3. In conclusion, the present study revealed that miR‑802 may suppress cervical cancer progression by decreasing BTF3 expression levels, indicating that it may represent a potential therapeutic target for the treatment and prognosis of patients with cervical cancer.

背景与目标： : 已发现MicroRNA (miR)-802与多种类型的肿瘤的发生和发展有关; 但是，关于mir-802在宫颈癌中的作用的研究有限。因此，本研究旨在探讨mir-802在宫颈癌细胞中的调节作用。使用逆转录定量 (rt ‑ q)PCR分析宫颈癌组织和细胞中mir-802的表达水平，使用双报告荧光素酶活性测定法鉴定mir-802的直接靶基因，进行rt ‑ qpcr和western blotting以确定mir-802和碱性转录因子3 (BTF3) 之间的关系。细胞活力、迁移和侵袭分别使用细胞计数kit-8和Transwell检测进行分析。最后，转移相关蛋白、n-钙粘蛋白和e-钙粘蛋白的表达水平，使用rt ‑ qpcr和western blotting进行测定。在宫颈癌组织和细胞中发现mir-802的表达水平降低，并且mir-802的过表达抑制了细胞活力，迁移和侵袭。此外，发现mir-802直接靶向BTF3以抑制其表达。值得注意的是，超表达mir-802显著逆转了BTF3对细胞活力的促进作用，同时，mir-802的过表达通过调节BTF3显著抑制上皮间质转化，以及细胞中基质金属肽酶 (MMP)2和MMP9的表达水平。总之，本研究表明，mir-8 5月02日通过降低BTF3表达水平来抑制宫颈癌的进展，表明它可能是宫颈癌患者治疗和预后的潜在治疗靶标。

BACKGROUND & AIMS: :This study aimed to identify the roles of the long non-coding RNA LINC00114 in colorectal cancer (CRC) development. The expression levels of LINC00114 and miR-133b in CRC were determined by reverse transcription (RT)-polymerase chain reaction (PCR) and the functions of LINC00114 in CRC were evaluated in vitro and in vivo. Methylation-specific PCR assay was performed to detect the miR-133b promoter methylation in CRC cells. Bioinformatics analysis, RNA immunoprecipitation, dual luciferase assay, RNA pull-down, co-immunoprecipitation (IP), and chromatin IP (ChIP) assays were used to elucidate whether LINC00114 could recruit EZH2/DNMT1 and bind to the miR-133b promoter region, leading to dysregulated methylation and the depression of miR-133b. The expression levels of DNA methyltransferases (DNMTs), EZH2, and nucleoporin 214(NUP214) were analyzed by western blotting. Data showed that LINC00114 was highly expressed, whereas miR-133b was downregulated in the CRC tissues and cells. In vitro, silencing LINC00114 inhibited cell proliferation and impeded cell cycle at the G1/S phase by upregulating miR-133b. In vivo, LINC00114 knockdown reduced tumor growth. Further analysis showed that the methylation in miR-133b promoter region was increased in the CRC and silencing LINC00114 increased miR-133b expression through depressing methylation of its promoter region. ChIP-PCR experiments demonstrated that EZH2 and DNMT1 could bind to the miR-133b promoter region and it was abolished by LINC00114 knockdown. sh-EZH2 reversed the overexpression of DNMTs and CRC cell cycle progression induced by the LINC00114 upregulation. LINC00114 could regulate the NUP214 protein expression by sponging miR-133b. These results demonstrated that LINC00114 suppressed miR-133b expression via EZH2/DNMT1-mediated methylation of its promoter region, indicating that LINC00114 might be a potential novel target for CRC diagnosis and treatment.

背景与目标： : 这项研究旨在确定长链非编码RNA LINC00114在结直肠癌 (CRC) 发展中的作用。通过逆转录 (RT)-聚合酶链反应 (PCR) 测定LINC00114和miR-133b在CRC中的表达水平，并在体外和体内评估LINC00114在CRC中的功能。进行甲基化特异性PCR测定以检测CRC细胞中的miR-133b启动子甲基化。使用生物信息学分析，RNA免疫沉淀，双荧光素酶测定，RNA下拉，共免疫沉淀 (IP) 和染色质IP (ChIP) 测定来阐明LINC00114是否可以募集EZH2/DNMT1并结合到miR-133b启动子区域，导致甲基化失调和miR-133b抑制。通过蛋白质印迹分析DNA甲基转移酶 (DNMTs)，EZH2和核孔蛋白214(NUP214) 的表达水平。数据显示LINC00114高表达，而miR-133b在CRC组织和细胞中被下调。在体外，沉默LINC00114通过上调miR-133b抑制细胞增殖并阻碍G1/S期的细胞周期。在体内，LINC00114敲低减少肿瘤生长。进一步的分析表明，miR-133b启动子区域的甲基化在CRC中增加，沉默LINC00114通过抑制其启动子区域的甲基化而增加miR-133b表达。ChIP-PCR实验表明EZH2和DNMT1可以结合miR-133b启动子区域，并被LINC00114敲低取消。sh-EZH2逆转了由LINC00114上调诱导的DNMTs的过表达和CRC细胞周期进程。LINC00114可以通过海绵miR-133b调节NUP214蛋白的表达。这些结果表明LINC00114通过其启动子区域的EZH2/DNMT1-mediated甲基化抑制miR-133b表达，表明LINC00114可能是CRC诊断和治疗的潜在新靶标。