Down-regulation of LncRNA CRNDE aggravates kidney injury via increasing MiR-181a-5p in sepsis

Long non-coding RNA (lncRNA) colorectal neoplasia differentially expressed (CRNDE) is reported to be linked to inflammation and cell apoptosis. However its role in sepsis induced kidney injury remains unclear. This study aims to explore the possible mechanism of CRNDE in kidney injury induced by sepsis. In vivo urine-derived sepsis (US) rat model and in vitro LPS-induced HK-2 and HEK293 cells were established. Kidney function was measured in rats from different groups. Relative levels of tumor necrosis factor-α (TNF-α) and interleukin-1β(IL-1β) in kidney tissue were detected via Enzyme-linked immune sorbent assay (ELISA). Then we up- or down-regulated CRNDE and miRNA-181a-5p expression in the cells. The biological influence of CRNDE and miR-181a-5p on cells was studied using CCK-8 assay and Annexin V assay. Interaction between CRNDE and miR-181a-5p was determined by bioinformatics analysis, RT-PCR, and dual luciferase reporter assay. Peroxisome proliferator-activated receptor-α (PPARα) and cell apoptosis related molecules were detected by western blot. We demonstrated that CRNDE was markedly down-regulated while miR-181a-5p was significantly up-regulated in sepsis models. CRNDE interacted with miR-181a-5p, and negatively regulated its expression level. CRNDE knockdown in rats increased the urea nitrogen and serum creatinine in plasma. Knockdown of CRNDE or transfection of miR-181a-5p significantly inhibited proliferation and promoted apoptosis of HK-2 and HEK293 cells, while overexpression of CRNDE and transfection of miR-181a-5p inhibitors had opposite effects. For mechanism, miR-181a-5p directly targeted the 3′ untranslated region of PPARα, and depressed its protein level, and PPARα was regulated indirectly by CRNDE. We concluded that CRNDE protected renal cell from sepsis-induced injury via miR-181a-5p/PPARα pathway.     

Long non-coding RNA CRNDE deteriorates intrauterine infection-induced neonatal brain injury

BackgroundThis study aimed to test the hypothesis that long non-coding RNA (lncRNA) colorectal neoplasia differentially expressed (CRNDE) could exacerbate brain injury caused by intrauterine infection in neonatal rats.MethodsIntrauterine infection was induced in pregnant rats by lipopolysaccharide (LPS). After delivery, newborn rats with brain injury caused by intrauterine infection were randomly divided into control, control shRNA, and CRNDE shRNA groups. CRNDE expression in serum and amniotic fluid of pregnant rats and neonatal brain tissues were determined by quantitative real-time PCR (qRT-PCR). Morris water maze (MWM) task was used to test the spatial learning and memory ability. Histological examination and apoptosis detection were performed by hematoxylin and eosin (H&E) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, respectively. Immunohistochemistry was conducted to evaluate the activation of astrocytes and microglia.ResultsLncRNA CRNDE was highly expressed in serum and amniotic fluid of maternal rats and in brain tissues of offspring rats. Furthermore, shRNA-mediated CRNDE downregulation could rescue the spatial learning and memory ability, improve brain histopathological changes and cell death, and inhibit the activation of astrocytes and microglia caused by LPS.ConclusionCRNDE silencing possessed a cerebral protective effect in neonatal rats with brain injury caused by interauterine infection.     

CRNDE affects the malignant biological characteristics of human glioma stem cells by negatively regulating miR-186

The long non-coding RNA Colorectal neoplasia differentially expressed (CRNDE) is a novel gene that activated early in colorectal neoplasia, but it is also up-regulated in many other solid tumors. Herein, the function and underlying mechanism of CRNDE in regulating glioma stem cells (GSCs) were investigated. We found that CRNDE expression was up-regulated while miR-186 expression was down-regulated in GSCs. Overexpression of CRNDE could promote the cellular proliferation, migration, invasion and inhibit the apoptosis in GSCs. Overexpression of miR-186 exerted functions of inhibiting the proliferation, migration and invasion of GSCs and promoting apoptosis. And CRNDE decreased the expression levels of XIAP and PAK7 by binding to miR-186 and negatively regulating it. In addition, miR-186 binded to XIAP and PAK7 3′UTR region, and decrease the expression of them, thus regulating the expression levels of downstream target proteins such as caspase 3, BAD, cyclin D1 and MARK2. The in vivo effect of CRNDE and miR-186 showed that the tumor formation rate was minimum in tumor-bearing nude mice with the knockdown of CRNDE and the overexpression of miR-186. In conclusion, CRNDE played an oncogenic role of GSCs through the negative regulation of miR-186. Both CRNDE and miR-186 could be regarded as potential targets in the glioma therapy.     

lncRNACRNDE 在非小细胞肺癌组织中的表达及其临床意义

目的:探讨结直肠差异表达长链非编码RNA CRNDE (colorectal neoplasia differentially expressed)在非小细胞肺癌(NSCLC)组织标本中的表达及其临床意义.方法:收集2011年1月至2015年12月成都军区总医院行根治性或姑息性切除术的NSCLC患者137例,以相应癌旁组织和29例肺外伤非肺癌患者的正常肺组织作为对照,通过实时荧光定量PCR法检测lncRNACRNDE在癌组织及癌旁组织标本中的表达,分析其表达与患者临床病理特征及预后的关系.结果:lncRNA CRNDE在肺癌癌组织中的平均表达水平为5.283±0.245,与非肿瘤组织(1.098±0.082)比较,差异有统计学意义(t=14.59,P＜0.01).lncRNA CRNDE 在肺癌组织标本中的表达较癌旁组织明显增高.lncRNA CRNDE的表达与患者的年龄、性别、吸烟史、肿瘤大小、淋巴结转移及肿瘤分化无关(均P＞0.05);与疾病分期、远处转移、病理类型及生存状态明显相关(均P＜0.05).lncRNA CRNDE高表达水平与NSCLC中的病理类型相关(P＜0.05),在腺癌组的表达水平明显高于鳞癌及其他类型组的表达水平;此外,高表达lncRNACRNDE的患者的PFS为(20.00±1.72)个月,较低表达者(34.07±1.97)缩短,差异有统计学意义(x2=15.940,P＜0.01);低表达lncRNA CRNDE的患者的OS为(47.50±2.31)个月,较高表达者(32.15±2.19)明显延长,差异有统计学意义(x2=12.40,P＜0.01).Cox多因素回归模型分析显示,lncRNA CRNDE的表达、远处转移及临床分期是NSCLC独立的预后因素(P＜0.05).结论:lncRNA CRNDE参与调节NSCLC的发生发展,可作为潜在的NSCLC诊断和预后评估的生物标志物.     

Highly expressed long non‐coding RNA CRNDE promotes cell proliferation through PI3K/AKT signalling in non‐small cell lung carcinoma

Recently, numerous studies have revealed that long non‐coding RNAs (lncRNAs) play complex roles in various lung diseases, while the colorectal neoplasia differentially expressed (CRNDE) functions in non‐small cell lung carcinomas (NSCLC) remain largely unknown. In the present study, we investigate the role and mechanism of CRNDE in the progression of NSCLC. The mRNA level of CRNDE in NSCLC patients and cells was detected by qRT‐PCR. The influence of CRNDE silencing or over‐expression on NSCLC cell proliferation and growth were assessed by MTT and flow cytometry, respectively. We also investigated the effect of abnormal CRNDE expression on cyclins and PI3K/AKT pathway. Furthermore, si‐CRNDE NSCLC cell lines were injected subcutaneously into nude mice to explore tumour formation in vivo. The expression of CRNDE was significantly upregulated in NSCLC patients and cells. In addition, both loss and gain function assays revealed that CRNDE promoted NSCLC cell proliferation and growth both in vitro and in vivo. Moreover, CRNDE regulated the cell cycle transition from G₀/G₁ stage to S stage and modulated the expression of CDK4, CDK6 and CCNE1. We further illustrated that CRNDE activated PI3K/AKT signalling in NSCLC cell lines. In conclusion, CRNDE was highly expressed in NSCLC malignant tissues and the heightened CRNDE strongly promoted NSCLC cell proliferation and growth through activating PI3K/AKT signalling; our results shed a light on utilizing CRNDE as a potential novel therapeutic target for the treatment of NSCLC.     

LncRNA CRNDE靶向miR-384影响结直肠癌细胞放射敏感性的研究

目的 研究长链非编码RNA （Lnc RNA）结直肠肿瘤差异表达基因（CRNDE）对结直肠癌细胞放射敏感性的影响及其机制。方法 以结直肠癌HT-29细胞作为体外研究对象,转染CRNDE shRNA,实时定量PCR测定干扰效果。以8 Gy X射线照射转染CRNDE shRNA后的HT-29细胞,四甲基偶氮唑盐（MTT）比色法和流式细胞术分别检测细胞增殖和凋亡水平。平板克隆实验检测放射敏感性。生物信息学软件预测CRNDE与miR-384有互补结合位点,荧光素酶报告系统鉴定靶向关系。将CRNDE shRNA和miR-384 inhibitor共转染至HT-29细胞中,以8 Gy剂量照射处理,MTT和流式细胞术检测细胞增殖和凋亡变化。结果 CRNDE shRNA能够降低HT-29细胞中CRNDE表达水平（1.00±0.08 vs. 0.42±0.06,t=10.051,P<0.05）。CRNDE shRNA和放射均可以抑制HT-29细胞增殖并诱导细胞凋亡,并且二者联合具有协同作用[凋亡率：（2.27±0.13）%、（23.58±2.35）%、（26.91±2.81）%、（36.84±3.24）%,F=24.66,P<0.05;吸光度（A）值：0.45±0.06、0.30±0.02、0.28±0.03、0.20±0.02,F=106.21,P<0.05]。CRNDE shRNA转染后可以提高HT-29细胞放射敏感性,放射增敏比为1.374。CRNDE靶向负调控miR-384表达。miR-384 inhibitor能够拮抗CRNDE shRNA对放射处理的结直肠癌细胞增殖抑制和凋亡促进的作用。结论 下调LncRNA CRNDE表达可增强结直肠癌细胞的放射敏感性,其作用机制与靶向负调控miR-384表达有关。     

长链非编码RNA CRNDE 通过调控NPHS1的表达促进糖尿病肾病足细胞损伤

目的 探讨长链非编码 RNA（lncRNA）在糖尿病肾病（DN）发生发展中的作用。方法 收集临床肾活组织检查标本,采用RNA-seq 测序技术检测DN组与正常对照组（NC组）肾组织中差异表达的lncRNA及mRNA。通过 GO、KEGG数据库分析差异表达mRNA的生物学功能,并通过共表达网络分析预测差异表达lncRNA的相互作用基因。采用实时荧光定量 PCR（qRT-PCR）检测目标lncRNA、mRNA 在DN肾组织中的相对表达水平。结果 RNA-seq 测序结果表明,与NC组相比,DN组共有353个差异表达的lncRNA,其中224个表达上调,129个表达下调。qRT-PCR 结果显示,DN组中CRNDE、PVT1和 BLZF2P 相对表达水平较NC组升高（P均< 0.001）,而WT1-AS、TARID 和 ST13P6相对表达水平较NC组降低（P均< 0.001）。共表达网络分析及双变量相关分析显示lncRNA CRNDE 与 NPHS1（编码的 nephrin 是足细胞结构完整性和发挥功能的决定性关键蛋白）呈负相关。结论 lncRNA CRNDE 在DN肾组织中表达明显升高,且与 NPHS1存在负相关关系,lncRNA CRNDE 可能通过调控 NPHS1 的表达促进DN足细胞损伤。     

长链非编码RNA CRNDE检测在乳腺癌诊断中的意义

目的 探讨长链非编码RNA(lncRNAs) CRNDE在乳腺癌患者血浆中的表达及临床意义。 方法 采用实时荧光定量聚合酶链反应方法检测76例乳腺癌患者组织、远端癌旁组织及血浆lncRNA CRNDE的表达情况。分析血浆中lncRNA CRNDE表达与乳腺癌的临床病理特征关系, 采用受试者工作特征曲线(ROC)评价血浆lncRNA CRNDE水平对乳腺癌的诊断效能。同时收集同期80例健康体检者为对照组检测其血浆中血浆lncRNA CRNDE表达情况。 结果 lncRNA CRNDE 在乳腺癌组织中的表达(2.73±1.66)明显高于癌旁组织(2.06±1.67),且在患者血浆中的表达水平(1.79±1.05)高于健康对照者(1.40±1.12),差异均有统计学意义(均P＜0.05);血浆中lncRNA CRNDE的表达水平与患者临床病理特征无关(P＞0.05);ROC曲线分析发现AUC为 0.66。 结论 血浆中高表达的lncRNA CRNDE可作为乳腺癌诊断的一个潜在的新型生物标志物。     

长链非编码RNA CRNDE调控胶质瘤细胞凋亡

目的 探讨长链非编码RNA CRNDE在胶质瘤细胞凋亡中发挥的功能和可能的作用机制.方法 将CRNDE基因干扰siRNA转染人胶质瘤U87细胞系,利用PE Annexin-V/7-AAD双染,流式细胞仪检测细胞凋亡情况,并利用Western方法检测细胞凋亡关键蛋白的表达变化.结果 siRNA干扰CRNDE表达的胶质瘤细胞中,细胞凋亡程度增加,凋亡相关蛋白Caspase3、Caspase7、Caspase9、PARP表达量明显升高.结论 CRNDE可能通过降低凋亡相关蛋白活性及表达水平,抑制胶质瘤细胞凋亡能力.