miR-369-5p抑制肝细胞癌细胞增殖、迁移及侵袭

目的:探讨miR-369-5p在肝细胞癌(hepatocellular carcinoma,HCC)中的表达情况、临床意义,观察miR-369-5 p对HCC细胞增殖、迁移及侵袭的影响及其作用机制.方法:实时定量PCR检测miR-369-5p在HCC组织与相应癌旁组织、正常肝细胞(L02)与HCC细胞系中的表达情况;C...     

Long noncoding RNA LINC02568 sequesters microRNA-874-3p to facilitate malignancy in breast cancer cells via cyclin E1 overexpression

Increasing numbers of long noncoding RNAs (lncRNAs) are implicated in breast cancer oncogenicity. However, the contribution of LINC02568 toward breast cancer progression remains unclear and requires further investigation. Herein, we evaluated LINC02568 expression in breast cancer and clarified its effect on disease malignancy. We also investigated the mechanisms underlying the pro-oncogenic role of LINC02568. Consequently, LINC02568 was upregulated in breast cancer samples, with a notable association with worse overall survival. Functionally, depleted LINC02568 suppressed cell proliferation, colony formation, and metastasis, whereas LINC02568 overexpression exerted the opposite effects. Our mechanistic investigations suggested that LINC02568 was physically bound to and sequestered microRNA-874-3p (miR-874-3p). Furthermore, miR-874-3p mediated suppressive effects in breast cancer cells by targeting cyclin E1 (CCNE1). LINC02568 positively controlled CCNE1 expression by sequestering miR-874-3p. Rescue experiments revealed that increased miR-874-3p or decreased CCNE1 expression recovered cell growth and motility functions induced by LINC02568 in breast cancer cells. In conclusion, the tumor-promoting functions of LINC02568 in breast cancer cells were enhanced by sequestering miR-874-3p and consequently over-expressing CCNE1. Our data may facilitate the identification of novel therapeutic targets in clinical settings.     

microRNA-627-5p inhibits colorectal cancer cell proliferation,migration and invasion by targeting Wnt2

BACKGROUND microRNA-627-5p(miR-627-5p) dysregulation has been observed in several cancer types, such as hepatocellular carcinoma, oral squamous cell carcinoma,glioblastoma multiforme, and gastric cancer. The biological function of miR-627-5p in colorectal cancer(CRC) growth and metastasis is yet unclear.AIM To investigate the effects of miR-627-5p on the malignant biological properties of colorectal malignant tumour cells by targeting Wnt2.METHODS The levels of miR-627-5p in colorectal tumour ti...     

长链基因间非编码RNA 00681竞争性结合miR-16促进黑素瘤细胞侵袭和迁移

目的:检测长链基因间非编码RNA 00681（long intergenic noncoding RNA 00681,linc00681）在恶性黑素瘤组织和皮肤良性痣组织中的表达,探索沉默linc00681对黑素瘤A375细胞侵袭和迁移能力的影响及机制。方法:采用一步法实时定量聚合酶链式反应（real time-quantitative polymerase chain reaction,RT-qPCR）检测恶性黑素瘤组织和A375细胞中linc00681的表达。利用小干扰RNA技术沉默黑素瘤A375细胞中linc00681的表达。应用划痕实验检测敲低linc00681对细胞迁移距离的影响,Transwell检测敲低linc00681对细胞迁移和侵袭能力的影响。生信分析网站分析linc00681和微小RNA-16（microRNA-16,miR-16）的潜在结合位点。荧光素酶报告实验检测linc00681与miR-16的结合能力。挽救实验检测miR-16对linc00681调控A375细胞迁移和侵袭能力的影响。结果:黑素瘤组织和A375细胞中linc00681的表达高于皮肤良性痣和正常黑素HEMa-LP细胞。沉默linc00681表达后,A375细胞迁移距离、迁移和侵袭细胞数目均显著降低。研究显示:linc00681可吸附结合miR-16;linc00681和miR-16共同沉默组侵袭和迁移能力明显高于linc00681单独沉默组。结论:linc00681在黑素瘤组织和A375细胞中表达升高,并可通过竞争性结合miR-16促进黑素瘤细胞迁移和侵袭。     

基因间长链非编码RNA152靶向微小RNA?103a?3p对非小细胞肺癌细胞增殖和侵袭迁移的影响

目的探讨基因间长链非编码RNA 152(LINC00152)靶向调控微小RNA-103a-3p(miR-103a-3p)表达及对非小细胞肺癌(NSCLC)细胞增殖和侵袭迁移的影响。方法采用实时定量PCR(QPCR)检测正常肺上皮细胞BEAS-2B及NSCLC细胞(ANIP-973、NCI-H157、A549和NCI-H1975)的LINC00152水平。选取LINC00152水平最高的细胞分别转染LINC00152特异性小干扰RNA(si-LINC00152组)或无关序列(si-NC组),另设未转染细胞为对照组。QPCR检测LINC00152水平,活细胞计数CCK-8法、Transwell小室和划痕实验测定细胞增殖、侵袭和迁移能力,Western blotting检测基质金属蛋白酶(MMP)-2、MMP-9和第10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN)的水平;荧光素酶报告实验验证LINC00152靶向结合miR-103a-3p的能力。结果NSCLC细胞的LINC00152水平均高于BEAS-2B细胞(P<0.05),尤其是NCI-H1975细胞的最高。si-LINC00152组的LINC00152水平为0.352±0.087,低于对照组的1.058±0.219和si-NC组的1.126±0.139(P<0.05)。与si-NC组和对照组相比,si-LINC00152组NCI-H1975细胞转染48、72 h的增殖活力下降(P<0.05);si-LINC00152组的划痕愈合率和穿膜细胞数分别为(27.386±2.428)%和(78.840±5.031)个,低于si-NC组的(77.675±4.803)%和(179.208±13.264)个及对照组的(76.371±5.385)%和(174.003±15.678)个(P<0.05);与si-NC组和对照组相比,si-LINC00152组的MMP-2和MMP-9水平均降低,而PTEN水平升高(P<0.05)。对照组和si-NC组上述指标的差异无统计学意义(P>0.05)。双荧光素酶报告分析证实,miR-103a-3p模拟物降低了野生型LINC00152的荧光素酶活性(P<0.05),但对突变型无影响(P>0.05)。结论LINC00152在NSCLC细胞中高表达并发挥促癌作用,与NSCLC的迁移侵袭密切相关,LINC00152与miR-103a-3p间的相互作用在NSCLC靶向治疗中有一定潜能。     

Long non-coding RNA small nucleolar RNA host gene 7 facilitates the proliferation,migration, and invasion of esophageal cancer cells by regulating microRNA-625

BackgroundEsophageal cancer (EC) is a highly aggressive malignant tumor, of which esophageal squamous cell carcinoma (ESCC) constitutes the main subtype. Long non-coding RNA (lncRNA) small nucleolar RNA host gene 7 (SNHG7) has been extensively studied in many tumors and has been confirmed to be an oncogene; however, it has yet to be investigated in an ESCC study. Therefore, this study intended to uncover the role of SNHG7 in ESCC.MethodsQuantitative real-time polymerase chain reaction was applied to measure the expression levels of SNHG7 and miR-625 in ESCC tumor tissues and cell lines. Cell Counting Kit-8 assay, 5-Ethynyl-2''-deoxyuridine assay, scratch assay, and Transwell assay were conducted to assess the proliferation, migration, and invasion ESCC cell. We verified the interaction between SNHG7 and miR-625 by performing the dual luciferase reporter gene experiment.ResultsCompared to that in adjacent normal tissues and HET1A cell lines, the expression level of SNHG7 in ESCC tumor tissues and ESCC cell lines was up-regulated, while the expression level of miR-625 was down-regulated. ESCC cell proliferation, migration, and invasion were significantly promoted by SNHG7 overexpression but inhibited by silencing of SNHG7. Further, luciferase reporter gene experiments confirmed that SNHG7 interacted with miR-625, and rescue experiments showed that SNHG7 promoted the malignant phenotype by inhibiting miR-625.ConclusionsSNHG7 is up-regulated in ESCC tumor tissues and cell lines, while miR-625 is expressed at a low level. SNHG7 is able to facilitate the proliferation, migration, and invasion of ESCC cells by targeting miR-625.     

Long noncoding RNA CCDC183-AS1 depletion represses breast cancer cell proliferation,colony formation,and motility by sponging microRNA-3918

Many studies have illustrated the significance of long noncoding RNAs in oncogenesis and promotion of breast cancer (BC). However, the biological roles of CCDC183 antisense RNA 1 (CCDC183-AS1) in BC have rarely been characterized. Thus, we explored whether CCDC183-AS1 is involved in the malignancy of BC and elucidated the possible underlying mechanisms. Our data confirmed elevated CCDC183-AS1 expression in BC, which was associated with poor clinical outcomes. Functionally, knocking down CCDC183-AS1 hampered cell proliferation, colony formation, migration, and invasion in BC. Additionally, the absence of CCDC183-AS1 restrained tumor growth in vivo. Mechanistically, CCDC183-AS1 executed as a competitive endogenous RNA in BC cells by decoying microRNA-3918 (miR-3918) and consequently overexpressing fibroblast growth factor receptor 1 (FGFR1). Furthermore, functional rescue experiments confirmed that inactivation of the miR-3918/FGFR1 regulatory axis by inhibiting miR-3918 or increasing FGFR1 expression could abrogate the CCDC183-AS1 ablation-mediated repressive effects in BC cells. In summary, CCDC183-AS1 deteriorates the malignancy of BC cells by controlling miR-3918/FGFR1 regulatory axis. We believe that our study can deepen our understanding of BC etiology and contribute to an improvement in treatment choices.     

前列腺癌PC3细胞的增殖和侵袭

目的：探究长链非编码RNA（long non-coding RNA, lncRNA）LINC00308对前列腺癌细胞的增殖、侵袭及迁移的影响及其相关作用机制。方法：利用基因芯片在前列腺癌组织与癌旁对照组织中筛选差异表达的 lncRNA 及 mRNA, 并确定LINC00308 及甲状腺激素受体因子 13（thyroid hormone receptor interactor13,TRIP13）为研究对象。MTT 实验、平板克隆及Transwell和划痕实验检测LINC00308对前列腺癌细胞增殖、侵袭及迁移能力的影响,应用裸鼠移植瘤在体内验证上述影响,应用Western blotting及免疫组化实验在瘤组织和癌细胞中探究LINC00308对TRIP13表达的影响。生物信息学分析技术与RNA免疫共沉淀（RNA immunoprecipitation, RIP）及qPCR和双荧光素酶基因报告实验预测并探究miR-361-5p与LINC00308及TRIP13之间的相互作用机制,并利用平板克隆、Transwell侵袭实验对癌细胞恶性生物行为进行验证。结果：芯片结果及qPCR共同证实LINC00308（P<0.01）与 TRIP13（P<0.05）在前列腺癌组织及 4 种细胞系中均异常高表达 ;细胞功能实验结果表明过表达LINC00308可以促进前列腺癌细胞PC3的增殖、侵袭及迁移能力（均P<0.05）,而下调前列腺癌细胞中LINC00308表达起相反作用。裸鼠移植瘤实验证实,LINC00308能够在体内促进前列腺癌PC3细胞的成瘤（P<0.05或P<0.01）,且能够在体内体外促进TRIP13 表达（P<0.05）。生物信息学分析与 RIP 及 qPCR 和双荧光素酶基因报告实验结果证实 miR-361-5p 能够分别与LINC00308与TRIP13的3''-UTR靶向结合,且LINC00308能够通过吸附miR-361-5p而作为内源性竞争RNA（competing endoge‐nous RNA,ceRNA）调控TRIP13的表达,MTT、平板克隆及Transwell实验检测癌细胞的增殖、克隆形成和侵袭能力变化证实了三者之间的调控作用。结论：在前列腺癌组织和细胞中异常高表达的LINC00308通过发挥ceRNA的功能抑制miR-361-5p表达而增强TRIP13表达,从而促进前列腺癌的增殖、侵袭及迁移能力。     

Upregulated in Hepatitis B virus-associated hepatocellular carcinoma cells,miR-331-3p promotes proliferation of hepatocellular carcinoma cells by targeting ING5

Hepatitis B virus (HBV) is a major risk factor for development and progression of hepatocellular carcinoma (HCC). It has been reported that viral infection can interfere with cellular microRNA (miRNA) expression and participate in the pathogenesis of oncogenicity. Our miRNAs array data indicated that miR-331-3p expression in HCC cell lines increased, but the relationship between miR-331-3p expression and HBV activity is unclear. Here, we observed elevated expression of miR-331-3p in different HCC cell lines expressing HBV. HBV, especially HBx, promotes miR-331-3p expression by enhancing its promoter activity. Using a miRNA target prediction database miRBase, we identified ING5 to be a novel target gene of miR-331-3p. miR-331-3p could inhibit ING5 expression by directly targeting its 3′-untranslated region (3′-UTR). As predicted, HBV was confirmed to repress ING5 at both mRNA and protein levels by promoting miR-331-3p expression. Our result indicated that miR-331-3p expression promotes proliferation of SMMC7721 cells by inhibiting ING5. ING5 overexpression promoted cell apoptosis in HCC cell lines. We also found ING5 expression was decreased in tumor tissue of HCC patient with HBV infection compared to its expression in para-carcinoma tissues. Conclusion: These results showed that miR-331-3p is upregulated by HBV and promotes proliferation of HCC cells though repression of ING5 expression. These data provide new insights for understanding the mechanisms of HBV-related HCC pathogenesis.     

长链非编码RNA SNHG1靶向miR-340-5p/CCND1轴调控食管癌细胞增殖、迁移和侵袭

目的:分析长链非编码RNA小核仁RNA宿主基因1（LncRNA SNHG1）靶向miR-340-5p/细胞周期蛋白1（CCND1）轴调控食管癌细胞增殖、迁移和侵袭。方法:体外培养人食管上皮细胞、食管癌细胞KYSE-30、TE-1、NEC、Eca109,qRT-PCR法测定细胞中LncRNA SNHG1、miR-340-5p、CCND1 mRNA水平。将对数期NEC细胞分为对照组、sh NC1组、sh LncRNA SNHG1组、sh NC2组、sh CCND1组、sh LncRNA SNHG1+miR-340-5p inhibitor组、sh CCND1+miR-340-5p inhibitor组。CCK-8法测定细胞增殖能力,Transwell小室法测定细胞侵袭、迁移能力,Western blot法检测CCND1、Ki-67、MMP-2蛋白表达,双荧光素酶验证miR-340-5p与LncRNA SNHG1、CCND1的靶向关系,通过裸鼠瘤内注射转染试剂进行体内试验。结果:与人食管上皮细胞相比,食管癌细胞KYSE-30、TE-1、NEC、Eca109中LncRNA SNHG1、CCND1 mRNA表达升高,miR-340-5p表达降低（P＜0.05）,其中NEC细胞变化最显著,所以使用NEC作为下续研究菌株。与对照组、sh NC组相比,sh LncRNA SNHG1组NEC细胞LncRNA SNHG1、OD450、侵袭细胞数、迁移细胞数、Ki-67、MMP-2降低（P＜0.05）;与对照组、sh NC组相比,sh CCND1组CCND1 mRNA与蛋白表达、OD450、侵袭细胞数、迁移细胞数、Ki-67、MMP-2表达降低（P＜0.05）。miR-340-5p与LncRNA SNHG1、CCND1均靶向结合,与sh LncRNA SNHG1组相比,sh LncRNA SNHG1+miR-340-5p inhibitor组OD450、侵袭细胞数、迁移细胞数、Ki-67、MMP-2蛋白表达升高（P＜0.05）;与sh CCND1组相比,sh CCND1+miR-340-5p inhibitor组OD450、侵袭细胞数、迁移细胞数、Ki-67、MMP-2蛋白表达升高（P＜0.05）;裸鼠移植瘤实验进行了体内验证。结论:LncRNA SNHG1沉默可能通过调控miR-340-5p/CCND1表达抑制食管癌NEC细胞增殖、侵袭与迁移,裸鼠体内也验证了这一结果。     

长链非编码RNA LOC101927476抑制卵巢癌细胞侵袭迁移和增殖

目的探讨长链非编码RNA(LncRNA)LOC101927476在卵巢癌细胞中的表达及其对卵巢癌生物学特征的影响。方法选取2018—2019年于中国医学科学院肿瘤医院手术治疗的卵巢癌患者。采用实时荧光定量PCR(real-time PCR)检测卵巢癌细胞系3AO、OVCA429、TOV21G、A2780、SKOV3以及22例卵巢癌原发瘤和转移瘤组织中LncRNA LOC101927476的表达水平, 采用TCGA数据库中卵巢癌转录组测序数据验证LncRNA LOC101927476和GATA4的表达, 采用慢病毒包装体系构建过表达LOC101927476(OE-LOC101927476组)和空载(OE-EV组)慢病毒并转染至3AO细胞。设计单链向导RNA(sgRNA), 利用CRISPR/Cas9构建LncRNA LOC101927476敲除细胞系。采用Transwell法和划痕实验检测LncRNA LOC101927476对卵巢癌细胞侵袭和转移能力的影响。采用细胞计数盒8(CCK-8)法检测细胞的增殖能力。结果 real-time PCR显示, 22例卵巢癌患者中, 20例转移灶中L...     

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。     

Long Noncoding RNA LINC01703 Exacerbates the Malignant Properties of Non-Small Cell Lung Cancer by Upregulating MACC1 in a MicroRNA-605-3p-Mediated Manner

Long intergenic nonprotein-coding RNA 1703 (LINC01703) has diagnostic significance in lung adenocarcinoma. However, its specific roles in non-small cell lung cancer (NSCLC) and downstream mechanisms have not been investigated. In the current study, we characterized the role of LINC01703 in NSCLC malignancy and elucidated its detailed mechanism of action. LINC01703 expression was measured by qRT-PCR. The regulatory effects of LINC01703 on the malignancy of NSCLC cells were assessed by multiple functional experiments. The targeted interaction was confirmed by RNA immunoprecipitation and luciferase reporter assays. Herein, overexpression of LINC01703 in NSCLC was indicated in the TCGA database and further proven in our cohort. Functional studies revealed that knocking down LINC01703 repressed cell proliferation, colony formation, migration, and invasion in vitro, which was accompanied by the induction of apoptosis. The tumor growth of LINC01703-silenced cells was also inhibited in vivo. Mechanistic analyses revealed that LINC01703 functioned as a competing endogenous RNA for microRNA-605-3p (miR-605-3p) in NSCLC cells, which thereby upregulated the miR-605-3p target metastasis associated with colon cancer 1 (MACC1). Rescue experiments highlighted that the regulatory actions of LINC01703 ablation on NSCLC cells were abolished in response to miR-605-3p downregulation or MACC1 overexpression. In conclusion, LINC01703 enhanced the aggressiveness of NSCLC cells by altering miR-605-3p/MACC1. Our work suggests the therapeutic potential of LINC01703/miR-605-3p/MACC1 in NSCLC.