沉默lncRNA UCA1通过上调miR-873-5p表达对胶质瘤细胞放射敏感性影响

目的 探讨lncRNA UCA1通过调控miR-873-5p表达对体外培养的胶质瘤SHG-44、U87、U251细胞放射敏感性的影响。方法 采用克隆形成实验检测X线照射0、2、4、6、8Gy SHG-44、U87、U251细胞存活情况,qRT-PCR检测SHG-44、U87、U251细胞中UCA1基因表达。以放射抵抗的U87、U251细胞为研究对象,沉默UCA1表达、过表达miR-873-5p后用克隆形成实验及流式细胞术检测细胞存活分数和细胞凋亡率。双荧光素酶报告基因实验和qRT-PCR检测验证UCA1和miR-873-5p的靶向关系。结果 UCA1在U87、U251细胞中表达上调。沉默UCA1或过表达miR-873-5p可抑制U87、U251细胞存活,并促进细胞凋亡。miR-873-5p是UCA1靶基因,UCA1可负性调控miR-873-5p1的表达。抑制miR-873-5p表达可逆转沉默UCA1对胶质瘤细胞放射敏感性的影响。沉默UCA1增加射线对胶质瘤细胞U251移植瘤的抑制作用。结论 沉默UCA1通过上调miR-873-5p表达,抑制胶质瘤细胞存活并促进其凋亡,从而提高胶质瘤细胞放射敏感性。     

lncRNA DNM3OS在喉鳞状细胞癌组织和细胞中的表达及其临床和生 物学意义

目的：检测lncRNA DNM3OS在喉鳞状细胞癌（laryngeal squamous cell carcinoma,LSCC）组织和LSCC细胞株中的表达及其临床意义,探讨其对LSCC TU177细胞体外增殖、迁移及侵袭的影响,并分析DNM3OS与EMT的关系。方法：从河北医科大学第四医院生物标本库选取2014年3月至2018年12月收治的68例LSCC患者手术切除的癌及癌旁组织标本,应用qPCR法检测DNM3OS在LSCC组织和细胞株中的表达水平。采用siRNA敲低TU177细胞中DNM3OS的表达,应用MTS、克隆形成及Transwell小室等方法分别检测敲低DNM3OS表达对TU177细胞增殖、迁移和侵袭等生物学行为的影响。应用qPCR和WB法检测转染si-DNM3OS后对EMT标志物上皮钙黏素（E-cadherin）、神经钙黏素（N-cadherin）、波形蛋白（vimentin）、扭曲蛋白（twist）、锌指转录因子2（SNAI2）mRNA和蛋白的变化。结果：LSCC组织中DNM3OS表达水平明显高于癌旁组织（P<0.01）,并与患者的TNM分期、淋巴结转移及生存期有关联（P<0.05 或 P<0.01）。DNM3OS在LSCC细胞株（Hep-2、AMC-HN-8、TU177、TU212及TU686）中均呈现不同程度的高表达（P<0.05 或 P<0.01）,转染si-DNM3OS后TU177细胞中DNM3OS的表达显著降低（P<0.01）。与对照组相比,DNM3OS表达敲低可抑制TU177细胞的体外增殖、迁移和侵袭能力（P<0.05 或 P<0.01）,可上调 TU177 细胞中E-cadherin的表达而下调 N-cadherin、vimentin、twist和SNAI2的表达（均P<0.01）。结论：DNM3OS高表达与LSCC的恶性进展有关,其可能为预测LSCC患者预后的潜在指标;DNM3OS可能通过影响EMT进程促进LSCC细胞的侵袭和转移。     

lncRNA HULC在膀胱癌组织中的表达及其对5637细胞恶性生物学行为的影响

目的:探讨长链非编码RNA(lncRNA)HULC在膀胱癌组织中的表达及其与患者临床病理特征的关系,以及沉默HULC对膀胱癌5637细胞增殖、凋亡、迁移与侵袭的影响。方法:选取2014年6月至2017年12月郑州人民医院手术切除的102例膀胱癌患者的癌及癌旁组织标本,以及人膀胱癌细胞株5637和人正常膀胱上皮细胞株SV-HUC-1,用qPCR法检测膀胱癌组织及细胞中HULC的表达,并分析HULC表达与膀胱癌患者临床病理特征的相关性,通过Kaplan-Meier生存曲线评估HULC对预后的影响。利用siRNA干扰技术将si-HULC、si-NC质粒转染进5637细胞中,分别采用CCK-8法、流式细胞术、划痕愈合实验及Transwell小室法检测沉默HULC对5637细胞增殖、凋亡、迁移及侵袭的影响。结果:膀胱癌组织中HULC的表达水平显著高于癌旁组织(P<0.01),HULC表达水平与患者肿瘤分级、肿瘤分期及淋巴结转移相关联(均P<0.05),HULC高表达患者的OS与PFS明显低于低表达者(均P<0.05)。5637细胞中HULC的表达水平明显高于SV-HUC-1细胞(P<0.01),沉默HULC后的5637细胞的增殖、迁移和侵袭能力均显著降低(均P<0.01)、细胞凋亡率明显升高(P<0.01)。结论:lncRNA HULC在膀胱癌组织及5637细胞中均高表达,沉默HULC表达可抑制膀胱癌细胞的增殖、迁移和侵袭能力并促进凋亡。     

lncRNA FAM95B1在胶质瘤中的表达及其影响LN382细胞增殖和迁移 的可能机制

目的：探讨lncRNA FAM95B1 对胶质瘤细胞增殖和迁移能力的影响并探究其相关作用机制。方法：选取2018年1月至2020年8月于合肥市第三人民医院行手术治疗的38例胶质瘤患者的胶质瘤组织及癌旁组织标本,利用qPCR检测胶质瘤组织与4种细胞系中FAM95B1的表达水平,以表达最低的胶质瘤LN382细胞为研究对象,转染空载质粒（对照组）或pcDNA3.1-FAM95B1质粒（实验组）。MTT法和划痕实验检测FAM95B1对LN382细胞增殖和迁移能力的影响。生物信息学分析技术和双荧光素酶基因报告实验预测并验证FAM95B1与miR-26a-5p及PTEN之间的相互作用机制,应用qPCR和WB法检测FAM95B1对miR-26a-5p和PTEN表达的影响。结果：FAM95B1在胶质瘤组织中的表达明显低于癌旁组织（P<0.01）。FAM95B1在多种胶质瘤细胞系中的表达均明显低于正常脑胶质细胞（均P<0.01）。过表达FAM95B1可以下调LN382细胞的增殖（P<0.05）和迁移能力（P<0.01）。FAM95B1 能够靶向结合 miR-26a-5p（P<0.01）,miR-26a-5p 能够靶向结合 PTEN mRNA（P<0.01）。过表达FAM95B1可下调LN382细胞中miR-26a-5p的表达（P<0.01）,促进PTEN mRNA的表达（P<0.01）。结论：在胶质瘤组织和细胞系中异常低表达的FAM95B1通过发挥竞争性内源RNA的功能抑制miR-26a-5p的表达而增强PTEN蛋白表达,抑制胶质瘤细胞系LN382的增殖和迁移。     

LncRNA在骨重建及骨相关疾病中的作用

骨重建可保持骨生物力学特性稳定,对维持骨强度具有重要意义。正常骨骼的生长和发育需要转录调控网络、信号传导通路、力学生物学及生物力学因素的紧密协调,而力学环境的改变,多种信号途径失调,影响骨的发育。长链非编码RNA(long non-coding RNA, lncRNA)是一类长度大于200 nt、生物来源广泛、2级及3级结构高度保守的RNA分子。研究显示,许多lncRNA参与骨骼系统的正常发育或平衡,调控成骨细胞的分化,以及参与骨肉瘤的发生。LncRNA表达失调与关节炎、骨质疏松症和骨癌等多种骨疾病密切相关,有望作为预测骨疾病的生物标志物。综述lncRNA的特征、参与骨重塑的lncRNA及其可能的作用机理,并讨论lncRNA作为生物标志物应用于治疗包括骨关节炎、骨质疏松症、骨癌等骨骼系统疾病的可能性,旨在为更好地理解和研究lncRNA在生物体内的作用提供参考。     

Long noncoding RNAs could be potential key players in the pathophysiology of Sjögren's syndrome

Long noncoding RNAs (lncRNAs) are a recently discovered class of noncoding functional RNAs encoded by metazoan genomes. Recent studies suggest a larger regulatory role for lncRNAs in critical biological and disease processes. Mounting evidence on the role of lncRNAs in regulating key processes of the immune system prompted us to hypothesize the role of lncRNAs as key regulators of the pathophysiology of Sjögren's syndrome (SS). We used two similar approaches based on reanalysis of microarray expression datasets and curation of lncRNA‐protein coding gene interactions from literature to derive support for our hypothesis. We also discuss potential caveats to our approach and suggest approaches to validate the hypothesis. Our analysis suggests the potential larger and hitherto unknown role of lncRNA regulatory networks in modulating the expression of key genes involved in the pathogenesis of SS and thereby modulating the pathophysiology of SS.     

Combined inhibition of EGFR and c-ABL suppresses the growth of triple-negative breast cancer growth through inhibition of HOTAIR

Triple-negative breast cancer (TNBC) does not express conventional therapeutic targets and is the only type of malignant breast cancer for which no designated FDA-approved targeted therapy is available. Although overexpression of epidermal growth factor receptor (EGFR) is frequently found in TNBC, the therapeutic effect of EGFR inhibitors in TNBC has been underwhelming. Here we show that co-treatment with clinically validated inhibitors of c-ABL (imatinib) and EGFR (lapatinib) results in synergistic growth inhibition in TNBC cells. The dual treatment leads to synergistic repression of the long non-coding RNA (lncRNA) HOTAIR (HOX Antisense Intergenic RNA). HOTAIR has been known to induce tumor growth and metastasis in breast cancer. Depleting HOTAIR alone phenocopies the dual treatment in growth suppression. We show that expression of HOTAIR is regulated by β-catenin through a LEF1/TCF4-binding site. The dual treatment blocks nuclear expression of β-catenin and prevents its recruitment to the HOTAIR promoter. Consistently, forced expression of β-catenin rescued HOTAIR expression and cell viability in the presence of both drugs. Upregulation of HOTAIR is associated with TNBC in cell lines and a cohort of primary tumors. This study elucidates a previously unidentified mechanism in TNBC linking signaling with lncRNA regulation which may be exploited for therapeutic gain.     

Upregulated lncRNA-UCA1 contributes to progression of hepatocellular carcinoma through inhibition of miR-216b and activation of FGFR1/ERK signaling pathway

The long non-coding RNA (lncRNA) urothelial carcinoma-associated 1 (UCA1) has been recently shown to be dysregulated, which plays an important role in the progression of several cancers. However, the biological role and clinical significance of UCA1 in the carcinogenesis of hepatocellular carcinoma (HCC) remain unclear. Herein, we found that UCA1 was aberrantly upregulated in HCC tissues and associated with TNM stage, metastasis and postoperative survival. UCA1 depletion inhibited the growth and metastasis of HCC cell lines in vitro and in vivo. Furthermore, UCA1 could act as an endogenous sponge by directly binding to miR-216b and downregulation miR-216b expression. In addition, UCA1 could reverse the inhibitory effect of miR-216b on the growth and metastasis of HCC cells, which might be involved in the derepression of fibroblast growth factor receptor 1 (FGFR1) expression, a target gene of miR-216b, and the activation of ERK signaling pathway. Taken together, our data highlights the pivotal role of UCA1 in the tumorigenesis of HCC. Moreover, the present study elucidates a novel lncRNA- miRNA-mRNA regulatory network that is UCA1-miR-216b-FGFR1-ERK signaling pathway in HCC, which may help to lead a better understanding the pathogenesis of HCC and probe the feasibility of lncRNA-directed diagnosis and therapy for this deadly disease.