LncRNA LINC00958通过靶向miR-490-3p促进甲状腺乳头状癌细胞的增殖、侵袭和迁移

目的:研究LncRNA LINC00958对甲状腺乳头状癌细胞的影响及其作用机制。方法:TPC-1和K-1细胞分别分成Control、shRNA-NC、shRNA-LINC00958-1、shRNA-NC+inhibitor-NC、shRNA-LINC00958-1+inhibitor-NC和shRNA-LINC00958-1+miR-490-3p inhibitor组。TPC-1和K-1细胞移植的荷瘤鼠分别分为shRNA-NC、shRNA-LINC00958-1、shRNA-NC+inhibitor-NC、shRNA-LINC00958-1+inhibitor-NC和shRNA-LINC00958-1+miR-490-3p inhibitor组。用实时荧光定量PCR(RT-qPCR)检测LINC00958和miR-490-3p的表达水平;CCK-8检测细胞活力;Western blot检测蛋白表达水平;细胞划痕实验检测细胞迁移水平,Transwell小室检测细胞侵袭水平,克隆形成实验检测细胞增殖水平;双荧光素酶报告实验检测LINC00958和miR-490-3p的靶向关系。测定肿瘤质量及体积。结果:相比于正常组织,LINC00958在甲状腺癌组织中高表达;相比于Nthy-ori 3-1细胞,LINC00958在TPC-1和K-1细胞中高表达,miR-490-3p低表达。LINC00958沉默后,甲状腺乳头状癌细胞的增殖、侵袭和迁移能力显著降低。干扰miR-490-3p表达逆转LINC00958沉默对甲状腺乳头状癌细胞的增殖、迁移和侵袭效果,破坏LINC00958沉默对肿瘤生长的抑制效果。结论:抑制LINC00958表达可抑制TPC-1和K-1细胞增殖、迁移和侵袭能力,其机制与靶向miR-490-3p表达有关。     

LINC01089通过调控miR-27a-3p/TET1轴抑制胰腺癌细胞的增殖和迁移

目的:本研究旨在探究胰腺癌中长链非编码RNA 01089（LINC01089）/miR-27a-3p/TET1轴在胰腺癌进展中的作用及其机制。方法:通过GEPIA数据库分析LINC01089在胰腺癌中的表达及其与预后的相关性。使用定量实时聚合酶链反应（qRT-PCR）检测LINC01089、miR-27a-3p、TET1 mRNA在胰腺癌组织和正常组织中的表达;CCK-8、BrdU和划痕愈合实验检测LINC01089和miR-27a-3p对细胞增殖和迁移的影响。机制上,我们通过StarBase和TargetScan数据库预测、双荧光素酶报告基因验证LINC01089和miR-27a-3p、miR-27a-3p和TET1之间的调控关系。采用Western blot检测LINC01089和miR-27a-3p对TET1表达的影响。结果:GEPIA数据库显示LINC01089在胰腺癌中低表达,其低表达与患者整体生存期(overall survival,OS)和无病生存期(diease free survival,DFS)相关。本研究中,我们发现与正常组织相比,癌组织中LINC01089表达显著下调,并与患者不良预后相关;功能实验显示LINC01089抑制胰腺癌细胞的增殖和迁移。相关机制实验表明LINC01089通过吸附miR-27a-3p上调TET1的表达在胰腺癌中发挥抑制作用。结论:LINC01089通过靶向调控miR-27a-3p促进TET1的表达,从而抑制胰腺癌细胞的增殖和迁移。     

TFESB调控lncRNA LINC00858/miR-324-5p通路抑制宫颈癌细胞的增殖、迁移和侵袭

目的:探讨半枝莲总黄酮提取物(total fiavonoids extract from Scutellaria barbata D.Don,TFESB)调控长链非编码RNA(lncRNA) LINC00858/miR-324-5p通路对宫颈癌细胞增殖、迁移和侵袭的影响及其机制.方法:qRT-PCR检测20例宫颈癌组织...     

lncRNA LINC000466 Predicts the Prognosis and Promotes the Progression of Triple‐negative Breast Cancer via Modulating miR‐539‐5p

BackgroundTriple-negative breast cancer (TNBC) is one of the most malignant subtypes of breast cancer with an unsatisfied prognosis. Effective biomarkers could predict the risk and improve patients’ survival. Whether LINC00466 possesses the potential to serve as a biomarker of the progression and prognosis of TNBC was evaluated.Materials and MethodsA total of 122 TNBC patients were included in this study and provided paired clinical tissues. The expression of LINC00466 in TNBC was evaluated by RT-qPCR and evaluated the clinical significance with a series of statistical analyses. The biological effects and the mechanism were investigated in TNBC cells with CCK8, Transwell, and luciferase reporter assays.ResultsLINC00466 was significantly upregulated in TNBC and showed close association with the clinical features of patients, which indicates the development, and severity of patients. LINC00466 functioned as a prognostic biomarker predicting the survival of patients and a tumor promoter improving the proliferation, migration, and invasion of TNBC cells through sponging miR-539-5p.ConclusionLINC00466 promotes the progression of TNBC via regulating miR-539-5p. The inhibition of LINC00466 might be a novel therapeutic strategy for TNBC.     

MicroRNA-183抑制结直肠癌肿瘤细胞的增殖、侵袭及迁移

目的:探讨microRNA183 （miRNA-183）对结直肠癌肿瘤细胞增殖、侵袭及迁移的影响。方法:使用慢病毒技术,分别转染HT29细胞株,建立稳定过表达细胞株miRNA-183 mimics,抑制miRNA-183表达的稳定细胞株miRNA-183 inhibitor以及阴性对照组negative control。qRT-PCR检测各组细胞miRNA-183的表达,Western blot检测miRNA-183下游靶基因PDCD4蛋白水平表达变化,CCK8增殖实验检测各组细胞的增殖情况,同时以划痕实验和Transwell实验分析转染miRNA-183 mimics和miRNA-183 inhibitor对HT29细胞迁移、侵袭能力的影响。结果:与阴性对照组相比,miRNA-183 mimics组细胞增殖速度明显下降,其迁移和侵袭能力也显著降低,差异均具有统计学意义（P<0.05）;miRNA-183下游靶基因PDCD4蛋白水平表达明显增加。结论:miRNA-183可有效地抑制结直肠癌肿瘤细胞的增殖、侵袭及迁移。     

LINC01503通过miR-342-3p/IGF2R轴促进上皮性卵巢癌的进展

目的：探讨LINC01503 在上皮性卵巢癌（EOC）中的表达水平和生物学功能及其可能的作用机制。方法：收集2015年5月至2016 年5月间在河北医科大学第四医院妇瘤科手术切除并经病理学确诊的85 例EOC患者的肿瘤组织和输卵管组织。常规培养人EOC 细胞A2780、SKOV3、OVCAR3 和OV90 及正常人卵巢上皮细胞IOSE80，将si-LINC01503、si-NC 及miR-342-3p mimic、miR mimic NC分别转染至SKOV3和A2780 细胞，分别作为si-LINC01503 组、si-NC 组、miR-342-3p mimic 组和miR mimic NC组。qPCR 法检测EOC组织和细胞中LINC01503 的表达水平，Kaplan-Meier 法分析LINC01503 表达水平与患者生存的关系。双荧光素酶报告基因实验验证LINC01503/miR-342-3p/IGF2R轴相关分子间的靶向关系。平板克隆、划痕愈合和Transwell 实验分别检测敲低LINC01503 及转染miR-342-3p mimic 对A2780 和SKOV3细胞增殖、迁移和侵袭能力的影响。WB法检测EOC细胞中LINC01503/miR-342-3p 通路对IGF2R蛋白表达的影响。构建A2780 细胞裸鼠移植瘤模型，观察敲低LINC01503 对移植瘤生长的影响。结果：EOC组织和细胞中LINC01503 表达水平分别显著高于输卵管组织和IOSE80 细胞（均P<0.01），LINC01503高表达组患者术后PFS 和OS 均显著短于LINC01503 低表达组患者（均P<0.01）。敲低LINC01503、转染miR-342-3p mimic 均可抑制EOC 细胞的增殖、迁移和侵袭能力（均P<0.01）。敲低LINC01503 可下调IGF2R的表达（P<0.01），这一现象可通过转染miR-342-3p inhibitor 挽救。敲低LINC01503 可抑制A2780 细胞裸鼠移植瘤的生长（P<0.01）。结论：在EOC 组织和细胞中呈高表达的LINC01503，与患者的不良预后密切相关，LINC01503可能通过吸附miR-342-3p影响IGF2R表达进而促进EOC的进展。     

LINC00511靶向miR-497-5p调控胃癌细胞增殖、迁移和侵袭及机制研究

目的:探讨LINC00511对胃癌细胞增殖、迁移和侵袭的影响及其作用机制。方法:将pcDNA、pcDNA-LINC00511、si-NC、si-LINC00511、miR-NC、miR-497-5p分别转染至MGC-803细胞中,分别记为pcDNA组、pcDNA-LINC00511组、si-NC组、si-LINC00511组、miR-NC组、miR-497-5p组;将si-LINC00511质粒分别与anti-miR-NC、anti-miR-497-5p共转染至MGC-803细胞中,分别记为si-LINC00511+anti-miR-NC组、si-LINC00511+anti-miR-497-5p组。实时荧光定量PCR（RT-qPCR）检测miR-497-5p和LINC00511表达水平;蛋白质印迹（Western Blot）法检测细胞周期素D1（cyclin D1,CyclinD1）、p21、基质金属蛋白酶2（matrix metalloproteinase 2,MMP2）、基质金属蛋白酶9（matrix metalloproteinase 9,MMP9）蛋白表达水平;四甲基偶氮唑盐比色法(MTT)检测细胞活性;Transwell检测细胞迁移和侵袭;双荧光素酶报告基因实验检测LINC00511和miR-497-5p的靶向关系。结果:与正常胃黏膜上皮细胞GES-1相比,胃癌细胞MGC-803、MKN-45、AGS中miR-497-5p表达水平显著降低,LINC00511表达水平显著升高。LINC00511靶向调控miR-497-5p的表达。抑制LINC00511表达和miR-497-5p过表达可降低细胞活性和迁移、侵袭数量,降低CyclinD1、MMP2、MMP9蛋白表达水平,提高p21蛋白表达水平。干扰miR-497-5p表达逆转了抑制LINC00511表达对胃癌MGC-803细胞增殖、迁移和侵袭的抑制作用。结论:抑制LINC00511表达可抑制胃癌细胞增殖、迁移和侵袭,其机制可能与miR-497-5p表达有关,将为胃癌的治疗提供新思路和新靶点。     

哈萨克族食管癌组织中microRNA-21的表达及其意义

目的：探讨哈萨克族食管癌组织中microRNA-21（miRNA-21）的表达及其意义。方法收集72例哈萨克族食管癌患者临床样本,采用实时定量PCR检测食管癌组织、周围正常食管组织miRNA-21表达水平;以患者性别、年龄、临床分期、肿瘤部位、分化程度、淋巴结转移、病情是否恶化等作为观察指标,分析miRNA-21与临床资料的关系。再以肿瘤中位生存期作为观察指标,筛选出影响哈萨克族食管癌患者预后的危险因素。结果食管癌组织miRNA-21相对表达量为（7.57±0.14）,周围正常食管组织相对表达量为（1.37±0.08）,食管癌组织相对表达量高于正常组织（P﹤0.05）。72例食管癌患者分为miRNA-21高表达组42例和低表达组30例,miRNA-21表达与临床分期、淋巴结转移、肿瘤进展有关,即淋巴结转移、肿瘤进展及肿瘤临床分期越晚,肿瘤miRNA-21表达也越高（P﹤0.05）;miRNA-21高表达组5年生存率为20.7%,低表达组5年生存率为38.6%,两组5年生存率比较差异具有统计学意义（χ2=4.715,P=0.005）;多因素分析显示,临床分期、肿瘤进展、miRNA-21表达水平是影响哈萨克族食管癌患者预后的独立危险因素（P﹤0.05）。结论 miRNA-21在食管癌组织表达水平高于正常组织,且miRNA-21表达越高,患者预后越差,miRNA-21可以作为哈萨克族食管癌诊断及预后评价的指标之一。     

Long Noncoding RNA LINC02163 Accelerates Malignant Tumor Behaviors in Breast Cancer by Regulating the MicroRNA-511-3p/HMGA2 Axis

Long intergenic nonprotein-coding RNA 02163 (LINC02163) has been reported to be upregulated and work as an oncogene in gastric cancer. The aims of the present study were to determine the expression profile and clinical value of LINC02163 in breast cancer. Additionally, the detailed functions of LINC02163 in breast cancer were explored, and relevant molecular events were elucidated. In this study, LINC02163 was upregulated in breast cancer, and its expression level was closely associated with tumor size, lymph node metastasis, and TNM stage. Patients with breast cancer presenting high LINC02163 expression exhibited shorter overall survival than those presenting low LINC02163 expression. Knockdown of LINC02163 resulted in a decrease in breast cancer cell proliferation, migration, and invasion and an increase in cell apoptosis in vitro. In addition, silencing of LINC02163 impeded breast cancer tumor growth in vivo. Mechanistic investigation revealed that LINC02163 served as a competing endogenous RNA for microRNA-511-3p (miR-511-3p) and consequently upregulated the expression of the high-mobility group A2 (HMGA2), a downstream target of miR-511-3p. Intriguingly, miR-511-3p inhibition and HMGA2 restoration counteracted the effects of LINC02163 deficiency on the malignant properties of breast cancer cells. LINC02163 exerts cancer-promoting effects during the initiation and progression of breast cancer via regulation of the miR-511-3p/HMGA2 axis. Our findings add to our understanding of the roles of the LINC02163/miR-511-3p/HMGA2 pathway as a regulator of breast cancer pathogenesis and may be useful in the development of lncRNA-directed cancer diagnosis, prognosis, and therapy.     

miR-885-5p通过靶向CTNNB1抑制胰腺癌细胞增殖、迁移和侵袭

目的:探讨miR-885-5p对胰腺癌细胞增殖、迁移和侵袭的影响,并阐明miR-885-5p和CTNNB1基因在胰腺癌细胞中的作用机制。方法:MTT、细胞划痕和Transwell实验测定各组细胞增殖、迁移与侵袭能力。双荧光素酶报告基因实验分析CTNNB1是否为miR-885-5p的靶基因。使用胰腺癌TCGA数据进行预后分析,并进行miR-885-5p与CTNNB1表达的相关性分析。结果:下调miR-885-5p明显促进胰腺癌细胞增殖、迁移与侵袭,而过表达miR-885-5p则相反。miR-885-5p可靶向CTNNB1 3' UTR并抑制其转录后表达。在下调miR-885-5p的细胞中进行回复性实验,结果表明,miR-885-5p对胰腺癌细胞增殖、迁移与侵袭的抑制作用部分依赖于CTNNB1。TCGA数据库结果显示,miR-885-5p高表达的胰腺癌患者有较好的预后,且与CTNNB1表达呈负相关。结论:miR-885-5p通过靶向CTNNB1抑制胰腺癌细胞增殖、迁移与侵袭,miR-885-5p有望成为胰腺癌治疗的新靶点。     

Hsa-miR-21-3p对食管癌细胞运动与侵袭能力的影响

目的： 研究hsa-miR-21-3p对人食管癌细胞Eca9706侵袭和迁移能力的影响。方法：采用终浓度为30 nmol/L的寡核苷酸hsa-miR-21-3p mimics和阴性对照转染Eca9706细胞48 h后,用实时荧光定量PCR检测转染后hsa-miR-21-3p的表达水平,通过Transwell体外侵袭试验和划痕愈合试验分别观察hsa-miR-21-3p对Eca9706细胞侵袭和迁移能力的影响。结果：与阴性对照组相比较,hsa-miR-21-3p mimics转染组细胞中hsa-miR-21-3p表达明显升高 (P<0.01),是阴性对照的1.1×104倍;Transwell体外侵袭试验结果显示镜下每个视野穿膜细胞数试验组 (76.67±6.11)较阴性对照组 (42.33±2.52)增加了81.1% (P<0.01),表明hsa-miR-21-3p的过表达显著增强食管癌细胞的侵袭能力;划痕愈合试验结果显示,hsa-miR-21-3p试验组的划痕宽度较对照组窄,在相同的时间点,试验组的划痕愈合能力均大于对照组 (P<0.01),说明转染了hsa-miR-21-3p mimics的试验组细胞的迁移能力大于对照组。结论：Hsa-miR-21-3p的高表达可以使人食管癌细胞Eca9706的侵袭和迁移能力增强,提示其可能参与了食管癌进程中的浸润和转移。     

lncRNA Xist/miR-215-5p下调LPAR4表达对乳腺癌细胞增殖与转移的影响

目的:探讨溶血磷脂酸受体4（LPAR4)在乳腺癌组织中的表达水平及与乳腺癌发生发展的关系,探究lncRNA Xist/miR-215-5p对LPAR4的调控作用。方法:通过Targetscan和Starbase预测LPAR4的上游通路lncRNA Xist/miR-215-5p。使用慢病毒转染LPAR4和miR-215-5p;使用质粒转染lncRNA Xist。在MDA-MB-231细胞系中,通过MTT和划痕实验检测细胞的增殖和转移能力。结果:LPAR4在乳腺癌细胞系中高表达,miR-215-5p和lncRNA Xist为低表达。Western blot显示LPAR4在乳腺癌细胞系中高表达。MTT和划痕实验提示上调LPAR4可以促进MDA-MB-231细胞的增殖和转移能力。进一步实验发现lncRNA Xist可以上调miR-215-5p,并且负性调控LPAR4;且lncRNA Xist可以抑制MDA-MB-231细胞的增殖能力。除此之外,在lncRNA Xist下调细胞中,上调miR-215-5p可以抑制lncRNA Xist下调介导的细胞增殖能力增强。结论:lncRNA Xist/miR-215-5p可能通过调控LPAR4的表达,为治疗乳腺癌提供可能的新靶点。LPAR4在乳腺癌细胞中高表达,可能成为乳腺癌潜在的肿瘤标志物。     

LINC00909靶向miR-365a-5p/FGFBP1分子轴调控结肠癌细胞增殖、迁移及侵袭的实验研究

目的 探讨LINC00909对结肠癌细胞增殖、迁移、侵袭的影响及可能机制.方法 购买正常结肠上皮细胞NCM460,结肠癌细胞HCT8、Caco-2及DLD-1;用实时荧光定量PCR(RT-qPCR)及蛋白质印迹法(Western blotting)检测NCM460细胞及结肠癌细胞中LINC00909、miR-365a-...     

lncRNA LINC00909靶向miR-548-3p对结直肠癌细胞放射敏感性的研究

目的 探讨lncRNA LINC00909是否通过靶向miR-548-3p而影响结直肠癌细胞放射敏感性。方法 采用qRT-PCR检测结直肠癌组织、癌旁组织中LINC00909、miR-584-3p的表达量;体外培养结直肠癌细胞SW480、SW620,分别将si-NC、si-LINC00909、miR-NC、miR-584-3p mimics、si-LINC00909与anti-miR-NC、si-LINC00909与anti-miR-584-3p转染至SW480、SW620细胞,用4 Gy照射细胞;克隆形成实验检测细胞存活分数及放射增敏比;MTT检测细胞增殖;Transwell小室实验检测细胞迁移及侵袭;双荧光素酶报告实验验证LINC00909、miR-584-3p的靶向关系。裸鼠皮下移植瘤实验检测干扰LINC00909表达或抑制miR-584-3p表达对照射后移植瘤重量的影响。结果 结直肠癌组织中LINC00909的表达水平显著升高(P<0.05),miR-584-3p的表达水平显著降低(P<0.05);干扰LINC00909表达或miR-584-3p过表达后细胞存活分数明显降低(P<0.05),放射增敏比分别为2.017、1.762,并可抑制增殖、迁移及侵袭(P<0.05);双荧光素酶报告实验证实LINC00909可靶向结合miR-584-3p;干扰LINC00909表达后移植瘤重量显著降低(P<0.05)。共转染anti-miR-584-3p后移植瘤重量显著升高(P<0.05)。结论 干扰LINC00909表达可通过上调miR-548-3p的表达而减弱结直肠癌细胞增殖、迁移及侵袭能力从而增强细胞放射敏感性。     

Radiosensitivity of lncrna linc00909 targeting mir-548-3p on colorectal cancer cells

Objective To investigate whether lncRNA LINC00909 affected the radiosensitivity of colorectal cancer cells by targeting miR-548-3p. Methods The expression levels of LINC00909 and miR-584-3p in colorectal cancer and adjacent tissues were detected by qRT-PCR. The colorectal cancer cells SW480 and SW620 were cultured in vitro, and transfected with si-NC, si-LINC00909, miR-NC, miR-584-3p mimics, si-LINC00909, and anti-miR-NC and si-LINC00909, and anti-miR-584-3p, respectively. The cells were irradiated with a dose of 4 Gy. The cell survival fraction and sensitization enhancement ratio (SER) were detected by clone formation assay. Cell proliferation was detected by MTT assay. Cell migration and invasion were assessed by Trans well chamber assay. The targeting relationship between LINC00909 and miR-584-3p was confirmed by dual luciferase reporter assay. The effect of interfering with the expression of LINC00909 or inhibiting the expression of miR-584-3p on the weight of the xenograft tumor after irradiation was evaluated by subcutaneous xenograft experiment in nude mice. Results The expression level of LINC00909 in colorectal cancer tissues was significantly up-regulated (P<0.05), whereas the expression level of miR-584-3p was significantly down-regulated (P<0.05). After interfering with the expression of LINC00909 or miR-584-3p overexpression, the cell survival fraction score was significantly reduced (P<0.05), the SERs were 2.017 and 1.762, and cell proliferation, migration and invasion were suppressed (all P<0.05). Dual luciferase reporter assay confirmed that LINC00909 could target and bind to miR-584-3p. After interfering with the expression of LINC00909, the weight of the transplanted tumor was significantly reduced (P<0.05), whereas the weight of the transplanted tumor was significantly increased after co-transfection with anti-miR-584-3p (P<0.05). Conclusion Interfering with the expression of LINC00909 can inhibit the proliferation, migration and invasion ability of colorectal cancer cells by up-regulating the expression of miR-548-3p, thereby enhancing the cell radiosensitivity.     

miR-586靶向TLR7调控食管癌细胞CaES-17的凋亡和增殖

目的:探讨miRNA调控食管癌细胞CaES-17的凋亡和增殖的潜在机制。方法:纳入2019年06月至2020年06月在我院接受治疗的食管癌患者6例,采集其食管癌组织和癌旁组织后抽取总RNA进行miRNA高通量测序。在食管癌细胞CaES-17中过表达食管癌组织和癌旁组织中的差异miRNA,检测食管癌细胞CaES-17的凋亡和增殖情况。通过TargentScan7.2在线分析和荧光素酶报告实验分析miRNA的靶标mRNA。结果:6例患者的食管癌组织和癌旁组织通过高通量测序发现hsa-miR-30d-5p、hsa-miR-6818-5p、hsa-miR-525-5p、hsa-miR-1909-3p、hsa-miR-212-5p、hsa-miR-586、hsa-miR-1286、hsa-miR-365b-3p、hsa-miR-30e-5p、hsa-miR-4317在食管癌组织和癌旁组织中的表达差异在8倍以上,并且食管癌组织均高于癌旁组织。干扰上述miRNA后,发现干扰miR-586能够抑制食管癌细胞CaES-17的增殖水平和迁移水平,提高凋亡水平。过表达miR-586后,食管癌细胞CaES-17的凋亡水平下降,增殖水平和迁移水平上升。TargentScan7.2在线分析和荧光素酶报告实验发现miR-586靶向TLR7的3' 端非编码区。敲低TLR7后,食管癌细胞CaES-17的凋亡水平下降,增殖水平和迁移水平上升。过表达TLR7后,食管癌细胞CaES-17的凋亡水平上升,增殖水平和迁移水平下降。但是,同时干扰miR-586和敲低TLR7后,相比于对照食管癌细胞,食管癌细胞CaES-17的凋亡水平和增殖水平无显著差异。结论:miR-586通过靶向TLR7的3' 端非编码区,降解了TLR7的mRNA,抑制了TLR7的蛋白翻译,最终抑制了食管癌细胞CaES-17的凋亡、促进了食管癌细胞CaES-17的增殖。