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。     

Long non-coding RNA NEAT1 regulates ferroptosis sensitivity in non-small-cell lung cancer

ObjectivesFerroptosis is caused by iron-dependent lipid peroxide accumulation, the sensitivity of which might be regulated by acyl-CoA synthetase long chain family member 4 (ACSL4). Non-small-cell lung cancer (NSCLC) can resist oxidative stress and reduce the sensitivity of tumor cells to ferroptosis by changing the expression of some proteins. Mechanisms involving ferroptosis sensitivity in NSCLC are not fully understood.MethodsA dual-luciferase reporter assay was used to confirm a targeting relationship between long non-coding (lnc)RNA NEAT1 and ACSL4. Overexpression and silencing assays of NEAT1 function were used to determine its roles in cell death (by TUNEL staining) and lipid peroxidation (by malondialdehyde levels). Expression of ferroptosis-related proteins (SLCA11, GPX4, and TFR4) was evaluated by western blot in NSCLC cells treated or not with the ferroptosis inducer erastin.ResultsErastin-induced cell death was positively correlated with ACSL4 level. NEAT1 regulated levels of ACSL4 and proteins related to the ferroptosis and classical apoptosis pathways. Levels of ACSL4, SLC7A11, and GPX4 were decreased more by NEAT1 silencing plus erastin than by erastin alone.ConclusionNEAT1 regulates ferroptosis and ferroptosis sensitivity, with the latter depending on ACSL4, suggesting that targeting NEAT1 or ACSL4 may be a viable therapeutic approach to the treatment of NSCLC.     

长链非编码RNA与肿瘤诊断

非编码RNA参与了多种疾病尤其是肿瘤发生发展的调控过程,是近期研究热点之一。随着高通量筛选方法的完善,越来越多的lncRNA分子被发现,并有望成为新型肿瘤诊断标志物和肿瘤治疗的靶点。近期研究提示IncRNA在肿瘤诊断和治疗方面具有良好的临床应用前景。本文介绍了lncRNA近期研究进展,相关lncRNA数据库的使用,并着重介绍了lncRNA与肿瘤诊断和预后关系研究情况。     

Correction: Long non-coding RNA MEG3 inhibits cell proliferation,migration, invasion and enhances apoptosis in non-small cell lung cancer cells by regulating the miR-31-5p/TIMP3 axis

Correction for ‘Long non-coding RNA MEG3 inhibits cell proliferation, migration, invasion and enhances apoptosis in non-small cell lung cancer cells by regulating the miR-31-5p/TIMP3 axis’ by Kui Li et al., RSC Adv., 2019, 9, 38200–38208, DOI: 10.1039/C9RA07880K.

In the published paper the Acknowledgements section was omitted; this should read:This work was supported by Major Scientific and Technological Projects of Guangzhou Science and Technology Plan Projects (No. 201802020004).The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers.     

Retracted Article: Long non-coding RNA MEG3 inhibits cell proliferation,migration, invasion and enhances apoptosis in non-small cell lung cancer cells by regulating the miR-31-5p/TIMP3 axis

Non-small cell lung cancer (NSCLC) is a malignant lung cancer and accounts for 80% of lung cancer-related deaths. Long non-coding RNA maternally expressed gene 3 (MEG3) has been identified as a tumor suppressor in multiple cancers. However, the regulatory mechanism of MEG3 in NSCLC development is still largely unknown. The expression levels of MEG3, microRNA-31-5p (miR-31-5p) and tissue inhibitor of metalloproteinase 3 (TIMP3) in NSCLC tumors and cells were measured by quantitative real time polymerase chain reaction (qRT-PCR). Cell viability, apoptosis, migration and invasion were detected by cell counting kit-8 (CCK-8), flow cytometry, western blotting and transwell assays, respectively. Xenograft mouse models were established by subcutaneously injecting NSCLC cells stably transfected with Lenti-pcDNA or Lenti-MEG3. The interaction between miR-31-5p and MEG3 or TIMP3 was validated by luciferase reporter and RNA immunoprecipitation (RIP) assays. MEG3 and TIMP3 levels were up-regulated, whereas miR-31-5p expression was down-regulated in NSCLC tumors and cells compared with normal tissues and cells. Overexpression of MEG3 repressed cell proliferation, migration and invasion, but induced apoptosis in NSCLC cells. More importantly, MEG3 effectively hindered tumor growth in vivo. Next, luciferase reporter and RIP assays confirmed the interaction between miR-31-5p and MEG3 or TIMP3. Pearson''s correlation coefficient revealed that miR-31-5p was inversely correlated with MEG3 or TIMP3. Rescue experiments indicated that MEG3 regulated TIMP3 expression by sponging miR-31-5p in NSCLC cells. Thus, MEG3 inhibited cell proliferation, migration and invasion, but enhanced apoptosis in NSCLC cells through up-regulating TIMP3 expression by regulating miR-31-5p, indicating novel biomarkers for the therapy of NSCLC.

Non-small cell lung cancer (NSCLC) is a malignant lung cancer and accounts for 80% of lung cancer-related deaths.     

Long non-coding RNA DLX6-AS1/miR-141-3p axis regulates osteosarcoma proliferation,migration and invasion through regulating Rab10

Long non-coding RNA (lncRNAs) DLX6-AS1 plays significant roles in various types of malignant tumors, including osteosarcoma (OS), the most prevalent primary malignant bone tumor. However, the role and mechanism of DLX6-AS1 have not been fully illuminated in OS. Here, we aimed to find a novel mechanism for DLX6-AS1 in regulating the development of OS through sponging microRNA (miRNA). According to the luciferase reporter assay, RNA immunoprecipitation and RNA pull-down assay, miRNA (miR)-141-3p can physically interact with DLX6-AS1 and Rab10. The expressions of DLX6-AS1 and Rab10 were upregulated and miR-141-3p was downregulated in OS tissues and cells (MG-63 and U2OS), as described by RT-qPCR and western blotting. Moreover, there was a negative correlation between the expression of miR-141-3p and either DLX6-AS1 or Rab10, and a positive correlation between DLX6-AS1 and Rab10. Functionally, cell proliferation, migration and invasion were evaluated by utilizing the MTT assay and transwell assays. As a result, DLX6-AS1 knockdown suppressed OS cell proliferation, migration and invasion in MG-63 and U2OS cells, which was abolished by the downregulation of miR-141-3p. Similarly, the upregulation of Rab10 not only promoted OS cell progression in vitro, but also blocked the inhibitory effect of miR-141-3p overexpression in OS cells. Notably, DLX6-AS1 knockdown could, in turn, reverse the promoting effect of Rab10 on OS cell progression. Xenograft experiments depicted that DLX6-AS1 knockdown restrained the tumor growth of MG-63 cells in vivo. In conclusion, the knockdown of DLX6-AS1 might suppress OS progression via sponging miR-141-3p and downregulating Rab10, suggesting a novel DLX6-AS1/miR-141-3p/Rab10 pathway in OS progression.

Long non-coding RNA (lncRNAs) DLX6-AS1 plays significant roles in various types of malignant tumors, including osteosarcoma (OS), the most prevalent primary malignant bone tumor.     

An epigenetically distinct breast cancer cell subpopulation promotes collective invasion

Tumor cells can engage in a process called collective invasion, in which cohesive groups of cells invade through interstitial tissue. Here, we identified an epigenetically distinct subpopulation of breast tumor cells that have an enhanced capacity to collectively invade. Analysis of spheroid invasion in an organotypic culture system revealed that these “trailblazer” cells are capable of initiating collective invasion and promote non-trailblazer cell invasion, indicating a commensal relationship among subpopulations within heterogenous tumors. Canonical mesenchymal markers were not sufficient to distinguish trailblazer cells from non-trailblazer cells, suggesting that defining the molecular underpinnings of the trailblazer phenotype could reveal collective invasion-specific mechanisms. Functional analysis determined that DOCK10, ITGA11, DAB2, PDFGRA, VASN, PPAP2B, and LPAR1 are highly expressed in trailblazer cells and required to initiate collective invasion, with DOCK10 essential for metastasis. In patients with triple-negative breast cancer, expression of these 7 genes correlated with poor outcome. Together, our results indicate that spontaneous conversion of the epigenetic state in a subpopulation of cells can promote a transition from in situ to invasive growth through induction of a cooperative form of collective invasion and suggest that therapeutic inhibition of trailblazer cell invasion may help prevent metastasis.     

Imatinib facilitates gemcitabine sensitivity by targeting epigenetically activated PDGFC signaling in pancreatic cancer

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长链非编码RNA DBH-AS1通过下调AKT1表达抑制胰腺癌进展

目的与背景 本研究旨在检测长链非编码RNA DBH-AS1在胰腺癌中的表达情况,探讨DBH-AS1在胰腺癌进展中的潜在分子作用。方法 采用实时定量PCR检测基因表达。细胞增殖、成克隆生长和迁移侵袭能力分别通过CCK-8、克隆形成和transwell检测。蛋白质水平用免疫印迹法测定。结果GEPIA数据库和定量PCR结果证实,DBH-AS1在胰腺癌组织中表达下调,与癌旁正常胰腺组织相比差异均有统计学意义(P 均＜0.05)。在胰腺癌肿瘤组织中DBH-AS1的低表达与肿瘤分化差、TNM分期晚期、淋巴结转移、以及预后不良(无瘤生存时间和总体生存时间)有关(P均＜0.05)。DBH-AS1基因敲除可促进胰腺癌细胞的增殖、克隆形成、迁移和侵袭能力,与对照组相比差异均有统计学意义(P均＜0.05)。机制研究表明,在胰腺癌中DBH-AS1通过下调AKT1表达抑制mTOR信号通路。结论DBH-AS1通过降低AKT1的表达抑制胰腺癌的进展。     

血清长链非编码RNA LSINCT5在乳腺癌中的表达与临床意义

目的探讨长链非编码RNA(long-noncoding RNA,lnc RNA)LSINCT5在乳腺癌患者血清中的表达水平及其临床应用价值。方法分别搜集90例乳腺癌患者、88例乳腺良性疾病患者及94例体检健康者血清标本,以及35例乳腺癌患者术前和术后血清标本;荧光定量PCR(q RT-PCR)法检测各组血清中LSINCT5的表达水平,并分析其与临床病理参数的相关性;采用电化学发光免疫测定法(ECLI)检测各组血清中CA153、CEA的表达水平,并进行多元Logistic回归分析;绘制ROC曲线评估血清LSINCT5对乳腺癌的诊断效能。结果乳腺癌组血清LSINCT5表达水平[1.45(0.76,3.16)]明显高于良性疾病组[1.08(0.66,1.45),H=3.188,P=0.004]和健康人对照组[1.21(0.80,1.44),H=2.626,P=0.026];术后血清中LSINCT5表达水平[0.85(0.49,1.31)]较术前明显降低[1.10(0.67,2.23),U=0.764,P=0.005];此外,血清LSINCT5水平与乳腺癌患者TNM分期、淋巴结转移、PR、和Ki-67有关(P0.05),而与年龄、病理类型、ER和Her2无关(P0.05);血清LSINCT5单独诊断乳腺癌的ROC曲线下面积(AUC~(ROC))为0.60,敏感性和特异性分别为39.7%和100%,均高于传统标志物CA153(AUC~(ROC)=0.59,25.0%,95.6%)和CEA(AUC~(ROC)=0.54,33.8%,82.4%),且3项指标联合后其诊断效能(AUC~(ROC)=0.66,45.6%,100%)高于各项单独检测。结论乳腺癌患者血清中LSINCT5高表达,可能作为乳腺癌诊断的一个潜在的生物学标志物。     

长链非编码RNA TTN-AS1通过miR-3928调节p38MAPK参与宫颈癌进展的机制研究

目的 探究长链非编码RNA(LncRNA)TTN-AS1通过miR-3928调节p38MAPK参与宫颈癌进展的机制.方法 分析肿瘤基因组图谱数据库中宫颈癌患者TTN-AS1水平与MAPK14水平和患者生存之间的关系.双荧光素酶报告验证靶向关系.通过转染miR-3928模拟物和TTN-AS1构建miR-3928和/或TT...     

长链非编码RNA ZFAS1在非小细胞肺癌患者组织中表达及作用

目的检测非小细胞肺癌(NSCLC)患者癌组织中锌指结构反义转录本1(zinc finger antisense 1,ZFAS1)表达水平,探讨ZFAS1在NSCLC进展中的生物学作用。方法实时荧光定量RT-PCR检测ZFAS1在NSCLC患者癌组织和癌旁组织中的表达水平。RNA干扰ZFAS1在A549细胞中表达,细胞计数和克隆形成实验检测细胞增殖情况,流式分析检测细胞周期和细胞凋亡,Transwell迁移和基质胶侵袭实验检测细胞迁移和侵袭能力,实时荧光定量RT-PCR检测Cyclin D1、Bcl2、N-cadherin、ZEB1、Slug和Twist基因表达水平变化。结果 ZFAS1在NSCLC患者癌组织中的平均表达水平[0.01(0.002,0.054)]较癌旁组织[0.002(0.001,0.012)]明显升高(Z=-2.638,P0.01)。ZFAS1基因敲减后,A549细胞增殖能力明显减弱(P0.01);A549细胞周期G1期比例升高,S期比例下降(P0.01);A549细胞凋亡比例明显增加(P0.01);A549细胞迁移和侵袭能力明显下降(P均0.01);A549细胞中Cyclin D1、Bcl2、N-cadherin、ZEB1、Slug和Twist基因表达水平均降低(P均0.05)。结论 ZFAS1在NSCLC患者癌组织中呈高表达。ZFAS1基因敲减诱导细胞周期阻滞、凋亡和抑制上皮-间质转变(EMT),减弱NSCLC细胞增殖、迁移和侵袭能力。     

PVT1 induces NSCLC cell migration and invasion by regulating IL-6 via sponging miR-760

Non-small-cell lung carcinoma (NSCLC) accounts for approximately 80% of lung cancers with a high metastatic potential. Elucidating the mechanism of NSCLC metastasis will provide new promising targets for NSCLC therapy and benefit its prognosis. Plasmacytoma variant translocation 1 (PVT1) has been proven to be overexpressed in NSCLC. Although the oncogenic role of PVT1 in NSCLC has been reported, its mechanism remains unclear. Here, we verified that the knockdown of PVT1 inhibited NSCLC cell migration and invasion, and that its inhibitory role on A549 cells and H1299 cells was antagonized by interleukin-6 (IL-6) treatment. The results revealed that PVT1 regulates IL-6 by sponging miR-760 and identified the binding site of miR-760 in the 3′-UTR of IL-6. In conclusion, a new mechanism was revealed, wherein PVT1 regulates NSCLC cell migration and invasion via miR-760/IL-6, suggesting PVT1/miR-760/IL-6 as promising prognostic biomarkers and therapeutic targets for NSCLC metastasis.