非小细胞肺癌细胞A549顺铂耐药潜在机制的全转录组测序分析

目的通过全转录组测序分析比较野生型A549细胞和顺铂耐药A549细胞(A549/DPP)表达谱的差别,揭示非小细胞肺癌(NSCLC)顺铂耐药潜在机制。方法首先建立A549/DDP细胞系,对A549和A549/DDP进行全转录组测序,分别对lnc RNA-seq,circ RNA-seq和mi RNA-seq数据进行差异表达以及功能富集分析(KEGG和GO分析)。然后进行全转录组数据联合分析以及ce RNA网络的构建。结果与A549细胞系相比,其中4517个lnc RNA、123个circ RNA以及145个mi RNA在A549/DDP细胞中有差异表达。显著富集在与癌症相关的通路上。mi RNA-circ RNA-lnc RNA-m RNA四元网络包含了12个mi RNA,4个circ RNA,23个lnc RNA和9个m RNA节点。经过拓扑学性质分析hsa-mi R-125a-5p和hsa-mi R-125b-5p是顺铂耐药相关的关键mi RNA。结论肿瘤坏死因子信号通路和p53信号通路参与了A549/DPP耐药机制。Hsa-mi R-125a-5p和hsa-mi R-125b-5p可能是逆转顺铂抗性的潜在靶点。     

ZEB2和PTEN在先天性巨结肠发生中的意义分析

目的研究ZEB2和PTEN在先天性巨结肠中的表达情况,探讨两者在先天性巨结肠发生中可能的调控关系。方法采用实时定量PCR和蛋白电泳技术检测64例先天性巨结肠狭窄段和扩张段中ZEB2和PTEN mRNA及蛋白表达情况。采用Pearson相关性检验分析ZEB2和PTEN在先天性巨结肠狭窄段和扩张段中表达的相关性。在体外细胞SH-SY5Y中应用ZEB2 siRNA干扰技术降低ZEB2表达,检测其对PTEN表达的影响,利用Transwell实验、CCK-8实验和流式细胞仪技术检测ZEB2对细胞迁移、增殖、周期和凋亡功能。结果 ZEB2和PTEN mRNA在先天性巨结肠狭窄段的表达均比扩张段显著增高(ZEB2:1.2 823±0.1 323 vs 0.987 7±0.124 9,P=0.007 3;PTEN:0.113 2±0.010 9 vs 0.045 9±0.005 8,P0.001)。ZEB2和PTEN在先天性巨结肠狭窄段和扩张段组织中蛋白表达与mRNA表达一致(ZEB2:0.709±0.035 vs 0.531±0.027,P=0.016 6;PTEN:0.466±0.047 vs 0.234±0.052,P=0.029 3)。ZEB2与PTEN mRNA表达在巨结肠狭窄段(r=0.48,P0.001)和扩张段(r=0.54,P0.001)中均呈显著正相关。干扰ZEB2mRNA表达后,体外细胞SH-SY5Y的PTENmRNA和蛋白表达显著下降,细胞增殖和迁移能力受到显著抑制。结论 ZEB2和PTEN在先天性巨结肠狭窄段中表达显著增加;ZEB2表达增加可能是PTEN通过竞争性内源性RNA机制调控后的继发性的改变。     

Analysis of long non‐coding RNA involved in atrazine‐induced testicular degeneration of Xenopus laevis

Long non‐coding RNA (lncRNA) plays a critical role in male germline development. Atrazine (AZ) as an environmental endocrine disrupting chemical (EDCs) can induce male reproductive toxicity in amphibians. Our previous studies demonstrated that AZ can alter gene and circular RNA (circRNA) expression of damaged testes in Xenopus laevis (X. laevis). We furthered to investigate the lncRNA expression profiling in the testis of X. laevis. Over 3559 lncRNAs were detected by lncRNA sequencing. AZ induced 40 upregulated and 46 downregulated differentially expressed lncRNAs. KEGG analysis showed that AZ‐affected lncRNAs mainly involve in 19 pathways among which 12 pathways are found in circRNA analysis. This study for the first time demonstrated that AZ can alter lncRNAs which may play a role in testicular degeneration through regulating expressions of functional genes in X. laevis. Our data may provide more insights on the mechanism about male reproductive toxicity of EDCs.     

circRNA作为ceRNA在结直肠癌中的研究进展

环状RNA(circRNA)是由前体mRNA反向剪接而产生的共价闭合RNA分子,在疾病中发挥着重要的调控作用。circRNA有潜力成为新型生物标志物,在肿瘤早期诊断、治疗和预后方面发挥重要作用。circRNA可以作为竞争性内源RNA(ceRNA),通过与miRNA相互作用,进而参与基因的转录后调控。本文重点概述了circRNA作为ceRNA调控结直肠癌发生发展的机制,以求能为结直肠癌的诊断与治疗拓展新视野。     

Comprehensive analysis of lncRNA-associated ceRNA network reveals the novel potential of lncRNA,miRNA and mRNA biomarkers in human rectosigmoid junction cancer

Although accumulating evidence has confirmed the potential biological functions of long non-coding RNAs (lncRNAs) as competitive endogenous RNAs (ceRNAs) in colorectal tumorigenesis and progression, few studies have focused on rectosigmoid junction cancer. In the present study, a comprehensive analysis was conducted to explore lncRNA-mediated ceRNA implications and their potential value for prognosis. lncRNA, microRNA (miR/miRNA) and mRNA expression profiles were downloaded from The Cancer Genome Atlas database. Subsequently, a lncRNA-miRNA-mRNA regulatory network was constructed to evaluate the functions of these differentially expressed genes on overall survival (OS) for rectosigmoid junction cancer. As a result, a rectosigmoid junction cancer-specific ceRNA network was successfully constructed with 7 differentially expressed (DE)lncRNAs, 16 DEmiRNAs and 71 DEmRNAs. Among the network, one DElncRNA (small nucleolar RNA host gene 20) and three mRNAs (sodium- and chloride-dependent taurine transporter, fibroblast growth factor 13 and tubulin polyglutamylase TTLL7) were significantly associated with OS (P<0.05). Additionally, two lncRNAs (KCNQ1OT1 and MIR17HG) interacted with most of the DEmiRNAs. Notably, two top-ranked miRNAs (hsa-miR-374a-5p and hsa-miR-374b-5p) associated networks were identified to be markedly associated with the pathogenesis. Furthermore, four DEmRNAs (caveolin-1, MET, filamin-A and AKT3) were enriched in the Kyoto Encylopedia of Gene and Genomes pathway analysis, as well as being included in the ceRNA network. In summary, the present results revealed that a specific lncRNA-miRNA-mRNA network was associated with rectosigmoid junction cancer, providing several molecules that may be used as novel prognostic biomarkers and therapeutic targets.     

LncRNA HIF1A-AS2 accelerates malignant phenotypes of renal carcinoma by modulating miR-30a-5p/SOX4 axis as a ceRNA

Objective: Several reports have proposed that lncRNAs, as potential biomarkers, participate in the progression and growth of malignant tumors. HIF1A-AS2 is a novel lncRNA and potential biomarker, involved in the genesis and development of carcinomas. However, the molecular mechanism of HIF1A-AS2 in renal carcinoma is unclear.Methods:The relative expression levels of HIF1A-AS2 and miR-30a-5p were detected using RT-qPCR in renal carcinoma tissues and cell lines. Using loss-of-function and overexpression, the biological effects of HIF1A-AS2 and miR-30a-5p in kidney carcinoma progression were characterized. Dual luciferase reporter gene analysis and Western blot were used to detect the potential mechanism of HIF1A-AS2 in renal carcinomas.Results:HIF1A-AS2 was upregulated in kidney carcinoma tissues when compared with para-carcinoma tissues (P < 0.05). In addition, tumor size, tumor node mestastasis stage and differentiation were identified as being closely associated with HIF1A-AS2 expression (P < 0.05). Knockdown or overexpression of HIF1A-AS2 either restrained or promoted the malignant phenotype and WNT/β-catenin signaling in renal carcinoma cells (P < 0.05). MiR-30a-5p was downregulated in renal cancers and partially reversed HIF1A-AS2 functions in malignant renal tumor cells. HIF1A-AS2 acted as a microRNA sponge that actively regulated the relative expression of SOX4 in sponging miR-30a-5p and subsequently increased the malignant phenotypes of renal carcinomas. HIF1A-AS2 showed a carcinogenic effect and miR-30a-5p acted as an antagonist of the anti-oncogene effects in the pathogenesis of renal carcinomas.Conclusions:The HIF1A-AS2-miR-30a-5p-SOX4 axis was associated with the malignant progression and development of renal carcinoma. The relative expression of HIF1A-AS2 was negatively correlated with the expression of miR-30a-5p, and was closely correlated with SOX4 mRNA levels in renal cancers.     

海南人咀嚼槟榔引起的口腔鳞状细胞癌ceRNA网络分析

目的:构建咀嚼槟榔引起的口腔鳞状细胞癌（OSCC）的ceRNA网络,探究其表达异常可能影响的调节通路改变。方法:对咀嚼槟榔的OSCC及癌旁组织进行miRNA测序和lncRNA微阵列分析,并建立了ceRNA网络。结果:lncRNA-miRNA-mRNA网络由308个lncRNA,41个miRNA和115个mRNA组成。结论:GO和KEGG分析表明,ceRNA网络参与了MAPK、凋亡、p53和PI3K/Akt等不同的信号通路。     

不同基因作为竞争性内源RNA参与膀胱癌调控的研究进展

竞争性内源RNA（competing endogenous RNA,ceRNA）是一种新的转录后RNA相互调控机制,越来越多的研究发现长链非编码RNA（long non-coding RNA,lncRNA）、环状RNA（circular RNA,circRNA）和假基因（pseudogenes）可以与微小RNA（microRNA,miRNA）竞争,影响靶RNA的稳定性或翻译,从而调控基因表达。近年来,不同基因作为ceRNA参与膀胱癌调控的研究已引起学者的广泛关注。为此,本文综述了lncRNA、circRNA和假基因作为ceRNA在膀胱癌中作用的研究进展,以及不同基因作为ceRNA调控膀胱癌的信号转导通路。从ceRNA分类的角度综述各类ceRNA在膀胱癌发生发展过程中的研究进展。     

Long noncoding RNA TMEM147-AS1 serves as a microRNA-326 sponge to aggravate the malignancy of gastric cancer by upregulating SMAD5

The abnormal expression of long noncoding RNAs (lncRNAs) is frequently observed in gastric cancer (GC) and considered an important driving force in GC progression. However, little is known regarding the involvement of TMEM147-AS1 in GC. Therefore, we examined TMEM147-AS1 expression in GC and determined its prognostic value. In addition, TMEM147-AS1 expression was depleted to identify the functional changes in response to TMEM147-AS1 deficiency. Using the cancer genome atlas dataset and our own cohort, we identified a strong expression of TMEM147-AS1 in GC. Increased TMEM147-AS1 levels in GC showed a significant association with poor prognosis. TMEM147-AS1 interference resulted in the inhibition of GC cell proliferation, colony-forming, migration, and invasion in vitro. Additionally, depletion of TMEM147-AS1 restricted the growth of GC cells in vivo. Mechanistically, TMEM147-AS1 functioned as a microRNA-326 (miR-326) sponge. Furthermore, SMAD family member 5 (SMAD5) was experimentally validated as the functional effector of miR-326. TMEM147-AS1 was demonstrated to sequester miR-326 away from SMAD5; consequently, knocking down TMEM147-AS1 downregulated SMAD5 levels in GC cells. The functional suppression of miR-326 or reintroduction of SMAD5 effectively reversed the attenuated behavior of GC cells caused by TMEM147-AS1 downregulation. In summary, TMEM147-AS1 exhibits tumorigenic activities in GC, which is likely the result of an altered miR-326/SMAD5 axis. Therefore, targeting TMEM147-AS1/miR-326/SMAD5 may represent a target for the treatment of GC.