The microarray identification circular RNA hsa_circ_0105015 up‐regulated involving inflammation pathway in essential hypertension

BackgroundEssential hypertension (EH) is an inflammatory disease, and endothelial dysfunction induced by chronic inflammation is one of the pathogeneses of EH. The expression of some inflammatory mediators may be regulated by the interaction of circular RNAs (circRNAs) and microRNAs (miRNAs).MethodsAn Agilent human circRNA microarray was used to identify the expression profile of circRNAs in EH. qRT‐PCR was used to evaluate the relative expression of circRNAs in 48 pairs of human whole blood samples (sex and age ± 3 years matched) and endothelial cells. TNF‐α was applied to induce endothelial cells inflammation. CircRNA‐miRNA network was predicted by MiRanda software.ResultsThere were 287 circRNAs differentially expressed in the microarray. The top 10 up‐regulated circRNAs in the EH group were hsa_circ_0014243, hsa_circ_0133228, hsa‐circRNA14116‐3, hsa_circ_0079536, hsa‐circRNA13649‐1, hsa_circ_0117886, hsa_circ_0007075, hsa‐circRNA15285‐1, hsa‐circRNA10088‐9, and hsa‐circRNA14119‐10; the top 10 down‐regulated circRNAs were hsa_circ_0100094, hsa_circ_0127342, hsa_circ_0093773, hsa_circ_0096334, hsa_circ_0131618, hsa_circ_0063886, hsa_circ_0097804, hsa_circ_0126640, hsa‐circRNA8935‐1, and hsa_circ_0039978 (fold change in descending order). Hsa_circ_0105015 has two predicted binding sites with hsa‐miR‐637. The relative expression of hsa_circ_0105015 in EH patients was significantly higher than healthy controls (P = .002), and similar results appeared in TNF‐α‐induced endothelial cells. The area under the curve after hsa_circ_0105015 combined with hsa‐miR‐637 was 0.703, P < .001.ConclusionHyperexpression of hsa_circ_0105015 is a significant risk factor of EH and its association with EH involves inflammatory pathways. Hyperexpression of hsa_circ_0105015 combined with hypoexpression of hsa‐miR‐637 indicates vascular inflammation or endothelial dysfunction and has potential as a biomarker for early diagnosis of EH.     

Circ-PRMT5 enhances the proliferation,migration and glycolysis of hepatoma cells by targeting miR-188-5p/HK2 axis

Introduction and objectivesCircular RNA (circRNA) has been demonstrated as a critical regulator in human cancer, including hepatocellular carcinoma (HCC). Nevertheless, the role of circ-PRMT5 in HCC remains largely unknown.Patients or materials and methodsThe real-time quantitative polymerase chain reaction (RT-qPCR) was performed to assess the expression levels of circ-PRMT5, miR-188-5p and anti-Hexokinase II (HK2) in HCC tissues and cells. The cell proliferation, migration and glycolysis were determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazol-3-ium bromide (MTT), transwell migration assay, and indicated kits, respectively. The interaction relationship between miR-188-5p and circ-PRMT5 or HK2 was analyzed by the bioinformatics database, dual-luciferase reporter assay, and RNA immunoprecipitation (RIP) assay. The western blot assay was used to analyze the expression level of HK2. The functional role of circ-PRMT5 in vivo was assessed by a xenograft experiment.ResultsCirc-PRMT5 was elevated in HCC tissues and cells than matched control groups. Furthermore, loss-of-functional experiments revealed that the silencing of circ-PRMT5 could repress proliferation, migration, glycolysis in vitro and tumor growth in vivo. Moreover, we also confirmed that overexpression of circ-PRMT5 abolished the effects on HCC cells induced by upregulating miR-188-5p. In addition, overexpression of miR-188-5p could repress the development of HCC. More importantly, HK2 was a target gene of miR-188-5p, and miR-188-5p regulated proliferation, migration, glycolysis of HCC cells by specifically binding to HK2. Mechanistically, circ-PRMT5 could act as a sponge of miR-188-5p to regulate the expression of HK2.ConclusionIn summary, circ-PRMT5 might play a key role in proliferation, migration, glycolysis of HCC cells via miR-188-5p/HK2 axis, which indicated that circ-PRMT5 might be a potential therapeutic target for HCC treatment.     

甲状腺癌RNA分子研究进展

甲状腺癌是内分泌系统中最常见的恶性肿瘤,约占全身恶性肿瘤的1%。目前在DNA水平研究甲状腺癌的发病机制已经相对成熟,但mRNA、miRNA、lncRNA、circRNA等不同RNA分子在甲状腺癌发病机制中的作用尚不清楚。已有研究表明,mRNA、lncRNA和circRNA等RNA分子共同构成动态的分子网络,主要包括lncRNA-miRNA-mRNA与circRNA-miRNA-mRNA两种调控网络,调节基因表达,参与肿瘤细胞的增殖、浸润、迁移和凋亡等过程,调节甲状腺癌的发生与进展。然而,不同类型的RNA分子在甲状腺癌中的具体作用机制尚不清楚。本文将重点对mRNA、miRNA、lncRNA、circRNA、ceRNA等RNA分子与甲状腺癌的关系进行综述,旨在深入阐明甲状腺癌发病相关RNA分子机制,从而为甲状腺癌的诊断和治疗提供新思路。     

环状RNA HIPK3在上皮性卵巢癌组织中的表达及其与临床病理和预后的关系探究

目的探究上皮性卵巢癌(EOC)组织中环状RNA(circRNA)HIPK3表达与临床病理及预后的关系。方法回顾性分析2011年5月至2014年12月于我院接受手术治疗的80例EOC患者的临床资料。收集所有EOC患者癌组织及对应癌旁正常组织(距癌灶边缘大于5 cm)标本,采用实时荧光定量PCR(qRT-PCR)检测EOC癌组织及癌旁正常组织中circRNA HIPK3的表达情况;根据癌组织中circRNA HIPK3表达水平的平均数分为高表达组(n=31)和低表达组(n=49),分析circRNA HIPK3表达与EOC的相关性;K-M生存曲线及Cox回归模型分析circRNA HIPK3与预后的关系。结果与癌旁正常组织相比,EOC癌组织中circRNA HIPK3表达显著降低(P<0.05);circRNA HIPK3表达量与肿瘤直径、分化程度、国际妇产科联盟(FIGO)分期及淋巴结转移显著相关(P<0.05);circRNA HIPK3高表达组术后5年累积总生存率显著高于低表达组(74.2%vs.38.8%,P=0.001);EOC分化程度(G3)[HR=1.48,95%CI(1.45,3.21)]、FIGO分期(Ⅲ~Ⅳ期)[HR=1.59,95%CI(1.53,3.47)]、淋巴结转移[HR=1.31,95%CI(1.28,2.46)]、术后未辅助化疗[HR=0.57,95%CI(0.45,0.97)]及circRNA HIPK3低表达[HR=0.24,95%CI(0.15,0.98)]是EOC患者预后不良的独立危险因素(P<0.05)。结论circRNA HIPK3低表达是EOC患者不良预后的独立危险因素之一,可作为新的临床肿瘤标志物及治疗靶点应用于临床。     

Hsa_circ_0078607在结直肠癌组织及血清中的异常高表达及其临床意义

目的：探究hsa_circ_0078607 在结直肠癌组织和患者血清中的表达水平及其与结直肠癌患者临床病理特征的关系,评价其能否作为结直肠癌潜在的分子诊断标志物及治疗靶标。方法：收集2018 年6月至2022 年1月于柳州市人民医院胃肠外科接受结直肠癌切除手术患者的58 对癌及癌旁组织标本,收集2020 年1月至2022 年12 月于柳州市人民医院初次确诊的结直肠癌患者、结直肠息肉患者及健康人体检血清共152 例;从结直肠癌差异表达circRNA 谱中挑选特异性高表达的hsa_circ_0078607作为候选标志物,采用qPCR法检测其在结直肠癌细胞、组织、患者血清及结直肠息肉患者血清中的相对表达量,分析其与临床病理特征的关系。采用ROC曲线评估hsa_circ_0078607 对结直肠癌及结直肠息肉的诊断价值。通过Circular RNA Interactome 数据库预测与hsa_circ_0078607 结合的miRNA,并用Cytoscape 3.9.1软件构建circRNA-miRNA-mRNA 调控网络,同时通过GO/KEGG富集分析进一步了解其功能。结果：与癌旁组织或健康人血清相比,hsa_circ_0078607 在结直肠癌细胞、组织和血清及息肉患者血清中呈高表达（P<0.001）,其中有52 例（89.7%）患者癌组织中表达上调,6 例（10.3%）表达下调。结直肠癌组织中hsa_circ_ 0078607 的相对表达量与肿瘤位置（P=0.029）、分化程度（P=0.046）和远处转移（P=0.043）有关联。ROC结果显示,在结直肠癌组织和血清中其诊断结直肠癌的AUC 分别为0.845 7[95%CI（0.772 8 ,0.918 6）,P<0.000 1]和0.868 3[95%CI（0.790 7,0.945 9）, P<0.000 1];在息肉患者血清中,hsa_circ_0078607 诊断结直肠息肉的AUC为0.710 1 [95%CI（0.610 0,0.810 1）]。GO/KEGG 富集分析结果表明,hsa_circ_0078607 下游的miRNA 可能参与RNA聚合酶Ⅱ启动子转录调控、蛋白K48-连锁泛素化、Wnt、Hippo 及MAPK信号通路调控等多个生物过程。结论：Hsa_circ_0078607 在结直肠癌细胞、组织和血清中呈高表达,其在结直肠癌组织中的表达水平与肿瘤位置、分化程度和远处转移有关联,提示其可作为结直肠癌潜在的分子诊断标志物;其还可能介导结直肠癌的发生发展过程,对发现结直肠癌潜在的治疗靶点有重要意义。     

雷公藤甲素抑制类风湿关节炎患者的成纤维样滑膜细胞的炎症和迁移： 基于circRNA 0003353/JAK2/STAT3信号通路

目的 探讨雷公藤甲素（TPL）通过调控环状非编码RNA（circRNA）0003353对类风湿关节炎（RA）成纤维样滑膜细胞（FLS）炎症和细胞迁移的作用机制。方法 纳入我院风湿科住院RA患者50例和正常人30例,收集外周血单个核细胞（PBMCs）及血清,检测circRNA 0003353的表达、免疫炎症指标[类风湿因子（RF）、C反应蛋白（CRP）、红细胞沉降率（ESR）、anti-CCP、IgA、IgG、IgM、C3、C4]和计算DAS28评分;采用最佳浓度10 ng/mL TPL处理RA-FLS,并转染circRNA 0003353过表达质粒。采用CCK-8法和Transwell实验检测RA-FLS细胞活力、迁移能力;ELISA法检测细胞因子IL-4、IL-6、IL-17;RT-qPCR法检测circRNA 0003353、Western blot检测p-JAK2、p-STAT3、JAK2、STAT3蛋白。结果 circRNA 0003353在RA患者PBMCs中表达升高,在协助诊断RA方面有良好效能（AUC=90.5%,P<0.001,95% CI：0.83-0.98）,circRNA 0003353与ESR、RF、DAS28呈正相关（P<0.05）;TPL呈时间依赖性降低circRNA 0003353表达,抑制RA-FLS的细胞活力和迁移能力,降低促炎细胞因子IL-6、IL-17,升高抑炎细胞因子IL-4（P<0.01）;TNF-α刺激后,RA-FLS中p-JAK2/JAK2、p-STAT3/STAT3比值升高,而TPL干预后降低（P<0.01）;在TPL干预基础上,转染circRNA 0003353过表达质粒,RA-FLS中circRNA 0003353表达升高,细胞活力和迁移能力升高,IL-4降低,IL-6、IL-17升高,p-JAK2/JAK2、p-STAT3/STAT3比值升高（P<0.01）。结论 circRNA 0003353在RA-PBMCs和RA-FLS中表达均升高,TPL可通过circRNA 0003353,调控JAK2/STAT3信号通路,抑制RA-FLS炎症和细胞迁移。     

环状RNA circUGGT2对结直肠癌细胞增殖、迁移和侵袭的影响

目的 探讨环状RNA circUGGT2在结直肠癌（colorectal cancer,CRC）中的表达及其对CRC细胞增殖、迁移和侵袭的影响。 方法 收集粤北人民医院胃肠外科2018年9月至2019年3月45例手术切除的CRC组织及相应癌旁组织标本,采用qRT-PCR检测组织及人正常结肠上皮细胞株NCM460和CRC细胞株HT29、HCT116、SW480、SW620中circUGGT2的表达水平。绘制受试者工作特征（ROC）曲线并计算曲线下面积（AUC）评估circUGGT2的诊断效能。将si-circUGGT2和si-NC分别转染至SW480细胞,采用CCK-8和平板克隆形成实验检测细胞增殖情况,划痕实验和Transwell小室实验检测细胞迁移和侵袭情况。结果 与癌旁组织比较,circUGGT2在CRC组织中高表达（P<0.001）,且与患者肿瘤大小、浸润深度和TNM分期有关（均P<0.05）;circUGGT2诊断CRC的AUC为0.702。circUGGT2在4种CRC细胞中的表达水平均高于NCM460细胞（均P<0.001）,尤其在SW480细胞中的表达水平最高。与si-NC组比较,沉默circUGGT2后SW480细胞增殖速率和划痕愈合速率降低（均P<0.05）,克隆形成数目及迁移、侵袭细胞数明显减少（均P<0.05）。结论 circUGGT2在CRC中高表达,沉默circUGGT2可抑制CRC细胞增殖、迁移和侵袭。circUGGT2可能是CRC诊断和治疗的潜在分子靶点。