miR-379对胶质瘤U87MG细胞增殖、迁移和侵袭能力的影响

目的研究miR-379对胶质瘤U87MG细胞增殖、迁移和侵袭能力的影响。方法 Real-time PCR方法检测miR-379在胶质瘤U87MG细胞和正常星型胶质细胞中的表达水平。在胶质瘤U87MG细胞中瞬时转染miR-379 agomir并用real-time PCR方法验证其转染效率。CCK-8方法检测miR-379对胶质瘤U87MG细胞增殖能力的影响。划痕实验检测miR-379对胶质瘤U87MG细胞迁移能力的影响。Transwell侵袭实验检测miR-379对胶质瘤U87MG细胞侵袭能力的影响。结果 miR-379在胶质瘤U87MG细胞中的表达水平显著低于在正常星型胶质细胞中的表达。miR-379 agomir抑制胶质瘤U87MG细胞的增殖、迁移和侵袭能力。结论 miR-379抑制胶质瘤U87MG细胞的增殖、迁移和侵袭能力。     

EMAP-Ⅱ上调miR-429表达抑制人U87胶质瘤细胞的增殖、迁移和侵袭能力

目的探讨内皮-单核细胞激活多肽-Ⅱ(EMAP-Ⅱ)对人U87胶质瘤细胞增殖、迁移和侵袭的影响以及可能的机制。方法培养人U87胶质瘤细胞,给予EMAP-Ⅱ处理不同时间后,采用CCK-8检测细胞活力的变化;应用real-time PCR方法检测mi R-429在人U87胶质瘤细胞中的表达变化;利用Lipofect AMINETM2000将antimi R-429及其阴性对照转染至人U87胶质瘤细胞,再给予EMAP-Ⅱ,分别应用CCK-8法和Transwell小室法检测人胶质瘤U87细胞的增殖、迁移及侵袭能力。结果与对照组相比,EMAP-Ⅱ显著抑制了人U87胶质瘤细胞的活力,上调mi R-429在人U87胶质瘤细胞的表达水平,上述变化在EMAP-Ⅱ作用0.5 h时效果最显著;与EMAP-II作用0.5 h组相比,转染anti-mi R-429后,EMAP-Ⅱ的作用被阻断,人胶质瘤U87细胞的细胞活力,以及迁移和侵袭能力基本恢复。结论 mi R-429参与了EMAP-Ⅱ抑制人U87胶质瘤细胞的增殖,迁移和侵袭的调控。     

miR-187靶向S100A4抑制胶质瘤U251细胞增殖、侵袭、迁移并诱导细胞凋亡

目的探讨miR-187对胶质瘤U251细胞增殖、侵袭、迁移和凋亡的影响及其机制。方法将体外培养的U251细胞分为Con组(未处理)、NC组(转染miR-NC)、miR-187组(转染miR-187 mimics)。采用实时定量PCR检测miR-187的表达,MTT法、Transwell小室实验和流式细胞仪分别检测细胞增殖、迁移、侵袭和凋亡,实时定量PCR和Western blot检测S100A4 mRNA和蛋白的表达,双荧光素酶报告基因实验检测miR-187和S100A4的靶向关系。结果与Con组相比,miR-187组细胞中miR-187表达水平和细胞凋亡率均明显升高,而细胞增殖活性、侵袭细胞数、迁移细胞数和S100A4 mRNA、S100A4蛋白的表达水平均明显降低(P0.05);而miR-NC组与Con组相比无显著性差异(P0.05)。双荧光素酶报告基因实验证实S100A4是miR-187的靶基因。结论 miR-187通过靶向S100A4抑制U251细胞增殖、迁移、侵袭并诱导细胞凋亡。     

circFOXM1靶向miR-6884-5p调控胶质瘤U251细胞增殖、迁移及侵袭的实验研究

目的：探讨circFOXM1对胶质瘤细胞增殖、迁移和侵袭的影响及可能机制。方法：RT-qPCR检测37例胶质瘤组织和正常脑组织中circFOXM1和miR-6884-5p表达;Pearson相关性分析验证胶质瘤组织中circFOXM1和miR-6884-5p表达的相关性;卡方检验分析胶质瘤组织中circFOXM1和miR-6884-5p的表达与患者临床病理特征的关系。体外培养胶质瘤U251细胞,分别转染si-circFOXM1、miR-6884-5p mimics或共转染si-circFOXM1与anti-miR-6884-5p后,CCK-8法和克隆形成实验检测细胞增殖;划痕实验和Transwell分别检测细胞迁移和侵袭,Western blot检测细胞中N-cadherin和E-cadherin蛋白表达。双荧光素酶报告基因实验验证circFOXM1和miR-6884-5p的调控关系。结果：胶质瘤组织中circFOXM1的表达量高于正常脑组织（P<0.05）,而miR-6884-5p的表达量低于正常脑组织（P<0.05）,两者呈负相关（r=-0.562 6,P<0.0...     

miR-107对人脑微血管内皮细胞增殖、迁移和血管形成能力的影响

目的探讨miR-107对人脑微血管内皮细胞增殖、迁移和血管形成能力的影响。方法miR-107化学模拟物agomir转染人脑微血管内皮细胞,CCK-8细胞活性分析实验和Transwell法分别检测内皮细胞增殖和迁移能力变化,体外血管生成实验检测内皮细胞成管能力的改变。结果内皮细胞转染miR-107化学模拟物agomir显著上调miR-107的表达。与阴性对照组(agomiR-NC组)相比,miR-107过表达组(agomiR-107)内皮细胞增殖和迁移显著减少,体外成管能力显著降低(P0.05)。结论miR-107能够抑制脑微血管内皮细胞增殖、迁移并降低其体外成管能力。     

miR-205-5p抑制体外胃癌细胞系增殖、迁移和侵袭

目的探讨miR-205-5p靶向RAS致癌家族基因2B(RAP2B)对胃癌细胞增殖、迁移和侵袭的影响。方法 RT-qPCR和Western blot检测胃癌细胞AGS、MGC803、MKN-28、SGC-7901和正常胃黏膜细胞GES-1中miR-205-5p和RAS致癌家族基因2B的表达。分别构建过表达miR-205-5p和抑制RAP2B表达的AGS细胞株,采用MTT法检测细胞活力;Transwell小室法检测细胞的迁移及侵袭能力;Western blot检测RAP2B、cyclin D1、MMP-2、MMP-9、GSK-3β和β-catenin蛋白的表达。采用双荧光素酶报告基因实验验证miR-205-5p对RAP2B的靶向作用。结果与正常胃黏膜细胞相比,4种胃癌细胞中miR-205-5p的表达显著降低,RAP2B的表达显著升高(P0.05)。过表达miR-205-5p或抑制RAP2B表达均可显著抑制AGS细胞的增殖、迁移和侵袭,抑制cyclin D1、MMP-2和MMP-9蛋白的表达(P0.05)。miR-205-5p可负性调控RAP2B的表达。结论 miR-205-5p通过靶向RAP2B抑制胃癌细胞的增殖、迁移和侵袭。     

miR-186介导YAP1调控乳腺癌细胞增殖、迁移和侵袭

目的 探讨miR-186介导的YAP1对乳腺癌细胞增殖、迁移和侵袭的影响。方法 采用qRT-PCR和Western blot法检测miR-186、YAP1蛋白在正常乳腺细胞MDA-kb2及人乳腺癌细胞MDA-MB-231中的表达;利用Lipofectamine 2000试剂将miR-186 mimic转染至乳腺癌细胞,荧光显微镜下观察转染效率;采用CCK-8法检测乳腺癌细胞的增殖能力;细胞划痕实验检测细胞的迁移能力;Transwell细胞侵袭实验检测细胞的侵袭能力;Western blot法检测YAP1蛋白表达。双荧光素酶报告基因实验检测miR-186与YAP1的靶向关系。结果 与正常乳腺细胞相比,乳腺癌细胞中miR-186表达量显著减少(P<0.01),YAP1蛋白表达量显著增加(P<0.01)。与对照组相比,miR-186过表达可明显降低乳腺癌细胞的增殖、迁移和侵袭能力(P<0.05),同时降低了YAP1蛋白表达(P<0.01)。miR-186和野生型YAP1载体共转染细胞的荧光素酶活性显著降低(P<0.01)。结论 miR-186在乳腺癌细胞中表达下...     

miR-342对胶质瘤细胞增殖,迁移侵袭及凋亡的影响

目的研究miR-342的表达变化对胶质瘤细胞生物学行为的影响。方法应用real-time PCR方法检测miR-342的内源性表达。CCK-8检测细胞增殖,Transwell法检测细胞的迁移侵袭,流式细胞仪检测细胞的凋亡。结果miR-342在胶质瘤细胞中表达水平较正常脑组织低;过表达miR-342能够抑制U87胶质瘤细胞的增殖、迁移侵袭,促进凋亡;相反,表达沉默miR-342能够促进U251胶质瘤细胞增殖、迁移侵袭,并抑制细胞凋亡(P0.05)。结论miR-342能够调控胶质瘤细胞的增殖、迁移侵袭,起抑癌基因的作用。     

七氟烷抑制人乳腺癌细胞系MCF-7增殖、迁移和侵袭

目的研究七氟烷对人乳腺癌细胞(MCF-7)增殖、迁移和侵袭的影响及其机制。方法用Western blot检测MCF-7细胞中高迁徙率族蛋白1 HMGB1的表达; miR-34a组(转染miR-34a mimics)、miR-con组(转染miR-con)、七氟烷处理+anti-miR-con组(转染anti-miR-con)及七氟烷处理+anti-miR-34a组(转染anti-miR-34a),均用脂质体法转染至MCF-7细胞; RT-qPCR检测细胞miR-34a表达; MTT法检测细胞增殖; Transwell小室法检测细胞迁移和侵袭;双荧光素酶报告基因实验检测细胞荧光活性。结果与对照组相比,1. 7%的七氟烷处理的细胞中miR-34a表达显著升高(P0. 05),HMGB1表达显著降低(P0. 05),且细胞增殖、迁移和侵袭均显著下调(P0. 05);过表达miR-34a可抑制MCF-7细胞增殖、迁移和侵袭,敲减miR-34a可降低七氟烷对MCF-7细胞增殖、迁移和侵袭的抑制作用。miR-34a可明显降低野生型HMGB1的细胞荧光活性,且负向调控HMGB1的表达。结论七氟烷可抑制人乳腺癌细胞增殖、迁移和侵袭。     

Knockdown of TMEM45A inhibits the proliferation,migration and invasion of glioma cells

Gliomas are the most common and aggressive type of primary adult brain tumor. Although high expression and prognostic value of TMEM45A has been recently reported in various types of human tumors, the association of TMEM45A expression and glioma is still unknown. Here, we reported that TMEM45A was significantly overexpressed in glioma tissues compared to non-tumorous brain tissues. Furthermore, TMEM45A mRNA levels were gradually increased with the increasing severity of histological grade of glioma. Moreover, high TMEM45A expression level was correlated with short survival time of glioma patients. Down-regulation of TMEM45A in two glioma cell lines, U251 and U373 by transected with TMEM45A siRNA resulted in a significant reduction of cell proliferation and G1-phase arrest. Additionally, we found that suppressing of TMEM45A expression in glioma cells remarkably suppressed cell migration and cell invasion. More importantly, TMEM45A siRNA treatment significantly down-regulated the proteins promoting cell cycles transition (Cyclin D1, CDK4 and PCNA) and cell invasion (MMP-2 and MMP-9), which indicted a possible mechanism underlying its functions on glioma. In summary, our study suggests that TMEM45A may work as an oncogene and a new effective therapeutic target for glioma treatment.     

miR-217调控lncRNAMALAT1影响食管鳞癌细胞的生物学行为

目的　探讨miR-217通过调控lncRNA MALAT1抑制食管鳞状细胞癌细胞的增殖,迁移和侵袭行为及其机制。 方法　qPCR检测miR-217在食管鳞状细胞癌组织和不同细胞株中的表达情况;双荧光素酶报告基因检测miR-217与MALAT1之间的相互作用;MTT增殖实验检测抑制miR-217后食管鳞状细胞癌细胞的增殖能力的变化情况;划痕愈合试验和Transwell侵袭实验检测抑制miR-217后食管鳞状细胞癌细胞的迁移和侵袭行为的变化情况;Western blotting实验检测miR-217对MALAT1下游相关蛋白表达情况的影响。 结果　与正常食管组织相比,食管鳞状细胞癌组织中miR-217的表达水平相对上调,与其他细胞株相比,Ec109细胞中miR-217表达最高;双荧光素酶实验证实miR-217能与MALAT1的3’ UTR特异性结合,可以调控MALAT1的表达与活性;抑制miR-217的表达后可以抑制食管鳞状细胞癌细胞的增殖、迁移和侵袭能力;抑制miR-217后MALAT1下游MIA2,ROBO1表达明显下调。 结论　miR-217可以调控MALAT1的表达影响食管鳞状细胞癌细胞的生物学行为。     

miR-107 promotes hepatocellular carcinoma cell proliferation by targeting Axin2

Background: A large number of studies demonstrated that microRNAs play important roles in the progression and development of human cancers. However, the expression level of miR-107 and its biological function in hepatocellular carcinoma (HCC) remains unclear. Method: Quantitative real-time PCR (qRT-PCR) was used to evaluate the expression level of miR-107 in HCC tissues and cell lines. Then, we explored the function of miR-107 to determine its potential roles on HCC cell proliferation in vitro. Luciferase reporter assay was used to confirm the target gene of miR-107, and the results were validated in cell lines. Results: miR-107 was significantly up-regulated in HCC tissues and cell lines. The enforced expression of miR-107 was able to promote cell proliferation in HepG2 cells. At the molecular level, our results suggested that expression of Axin2 was negatively regulated by miR-107. Conclusion: Our observations suggested that miR-107 could promote HCC cells proliferation via targeting Axin2 and might represent a potential therapeutic target for HCC.     

ATP1A1基因沉默抑制人胶质瘤细胞U251侵袭

目的探讨沉默Na+/K+ATP酶A1亚基(ATP1A1)对人U251胶质瘤细胞系侵袭能力的影响及其机制。方法用shRNA-ATP1A1慢病毒感染人U251胶质瘤细胞,RT-q PCR及Western blot分别检测ATP1A1 mRNA和蛋白的表达;MTT法检测细胞体外的增殖;细胞划痕实验及Transwell小室检测细胞的迁移及侵袭能力;Western blot检测基质金属蛋白酶2/9(MMP-2/9)的表达。结果沉默ATP1A1细胞的ATP1A1 mRNA和蛋白表达均受到了明显抑制;细胞的增殖和迁移、侵袭能力也显著受抑(P0.05);MMP-2和MMP-9的表达也明显降低(P0.05)。结论靶向ATP1A1干扰能够明显抑制胶质瘤U251细胞的体外增殖、迁移及侵袭,其机制可能与MMP-2、MMP-9的下调相关,ATP1A1可能作为胶质瘤治疗的一个潜在靶点。     

Adsorption of miR-218 by lncRNA HOTAIR regulates PDE7A and affects glioma cell proliferation,invasion, and apoptosis

Objective: To evaluate the role of targeted adsorption of miR-218 by long-chain non-coding RNAHOTAIR to regulate PDE7A on glioma cell proliferation, invasion, and apoptosis. Methods: The expressions of lncRNA HOTAIR, miR-218, and PDE7A in glioma tissues and normal parcancer tissues, NHA and glioma cell lines were determined, and correlations among the three genes were analyzed. The subcellular localization of lncRNA HOTAIR was determined by fluorescent in situ hybridization. Dual-luciferase reporter assay was used to validate the targeted relationship between lncRNA HOTAIR/miR-218/PDE7A. Glioma cells were grouped to receive intervention of lncRNA HOTAIR or miR-218. MTT, transwell, and flow cytometry were performed to determine the proliferation, invasion, and apoptosis of cells. Results: Compared with the normal tissues and cells, the expression of lncRNA HOTAIR was increased while miR-218 was suppressed in glioma tissues samples and cells (all P<0.05). Inhibition of lncRNA HOTAIR expression, was able to induce apoptosis and suppress the proliferation and invasion of cells (all P<0.05). LncRNA HOTAIR is mainly localized in the cytoplasm, and is able to adsorb miR-218 as ceRNA. The effect of knockdown of HOTAIR on glioma cells could be partially rescued by miR-218 inhibitor. The expression of PDE7A was enhanced in glioma tissues and cells compared to normal tissues and cells (all P<0.05), which positively correlated with the expression of HOTAIR (r=0.546, P<0.05) and negatively correlated with the expression of miR-218 (r=0.363, P<0.05). The targeted relationship between miR-218 and PDE7A was validated: Overexpression of miR-218 was able to suppress the proliferation and invasion of glioma cells and restrain apoptosis compared to the miR-NC group (all P<0.05). The effect of miR-218 on glioma cells could be partially rescued by PDE7A. Conclusion: lncRNA HOTAIR can adsorb miR-218 to regulate expression of PDE7A and promote the malignant biologic behavior of glioma cells.     

miR-124对C6胶质瘤细胞增殖和迁移能力的影响

目的:研究miR-124对C6胶质瘤细胞增殖和迁移能力的影响.方法:体外培养C6胶质瘤细胞,依据转染不同分为空白对照组、阴性对照组和miR-124拟似物组.实时荧光PCR检测转染效率,绘制生长曲线,MTT实验检测miR-124对C6细胞增殖能力影响,计算抑制率.划痕实验检测miR-124对C6细胞迁移能力影响,计算迁移率.结果:生长曲线显示miR-124拟似物组C6细胞生长能力受到明显抑制,转染后第3天,miR-124拟似物组C6细胞数目显著低于阴性对照组(5.410±0.463Vs.6.917±0.385;P＜0.01);miR-124拟似物组mRNA表达量显著高于阴性对照组和空白对照组(5.92±0.56 vs.0.93±0.13和1.00±0.12;P＜0.01);MTT增殖实验显示miR-124抑制C6细胞增殖能力,miR-124拟似物组抑制率显著高于阴性对照物组(42.90±5.169 vs.10.24±3.351;P＜0.01);划痕实验显示miR-124明显抑制C6细胞迁移能力,阴性对照组迁移率显著高于miR-124拟似物组(98.79±1.210vs.81.72±5.972;P＜0.05).结论:miR-124能够显著抑制C6胶质瘤细胞的增殖和迁移能力.     

负向调控miR-9抑制鼻咽癌细胞的增殖、迁移和侵袭*

 目的： 探讨负向调控miR-9对人鼻咽癌细胞增殖、迁移和侵袭作用。方法: 用脂质体LipofectamineTM 2000转染合成抑制剂的方法抑制鼻咽癌细胞miR-9表达,转染抑制对照剂作为对照组。CCK-8法和流式细胞术检测细胞增殖和细胞周期变化;Transwell侵袭实验和划痕实验检测细胞侵袭和迁移能力;免疫印迹实验检测蛋白变化。结果: 抑制鼻咽癌细胞miR-9表达后,肿瘤增殖能力降低（P<0.05）,G0/G1期细胞增多［CNE2:（57.96±1.39）% vs（47.93±1.76）%,P<0.05;CNE1:（51.24±0.88）% vs（48.29±0.39）%,P<0.05］,迁移距离明显缩短［CNE2:（186.50±7.94）μm vs （247.56±15.56）μm,P<0.05;CNE1:（139.06±16.73 ）μm vs（230.66±14.27 ）μm,P<0.01］,CNE2细胞中侵袭细胞数明显减少（43.00±3.17 vs 65.80±5.20,P<0.01）,β-连环蛋白（β-catenin）表达被抑制。结论: 在鼻咽癌细胞中,负向调控miR-9可抑制鼻咽癌细胞的增殖、侵袭和迁移。     

miR-197-3p调控DMBT1抑制甲状腺癌细胞系增殖、迁移和侵袭

目的探讨miR-197-3p是否通过靶向调控恶性脑瘤缺失1基因(DMBT1)影响甲状腺癌细胞增殖、迁移和侵袭。方法RT-qPCR检测健康人甲状腺细胞Nthy-ori 3-1和甲状腺癌细胞SW579、CGTHW-1中miR-197-3p表达;MTT法检测SW579细胞增殖;Transwell小室法检测SW579细胞迁移和侵袭;双荧光素酶报告基因实验验证miR-197-3p是否靶向DMBT1;Western blot检测细胞DMBT1、cyclinD1、p21、MMP-2和E-cadherin蛋白表达。结果与Nthy-ori 3-1细胞比较,SW579和CGTHW-1细胞中miR-197-3p相对表达量升高(P<0.05);抑制miR-197-3p表达后,SW579细胞的增殖、迁移和侵袭能力明显受到抑制,细胞中cyclinD1蛋白和MMP-2蛋白表达降低而p21蛋白和E-cadherin蛋白表达升高;SW579细胞中miR-197-3p靶向负调控DMBT1的表达;过表达DMBT1明显抑制SW579细胞增殖、迁移和侵袭,而抑制DMBT1则能逆转miR-197-3p对SW579细胞增殖、迁移和侵袭的影响。结论miR-197-3p通过靶向调控DMBT1的表达,抑制甲状腺癌细胞增殖、迁移和侵袭。     

MiRNA-15a inhibits proliferation,migration and invasion by targeting TNFAIP1 in human osteosarcoma cells

Recent data strongly suggest the important role of miRNAs in various cancer-related processes. Osteosarcoma is the most common type of primary malignant bone tumor and is characterized by complex genetic changes and resistance to conventional treatments. In this study, the role of miRNA-15a (miR-15a) in the progression and metastasis of osteosarcoma was investigated. The result demonstrated that the expression of miR-15a was down-regulated in osteosarcoma tissues and cell lines as compared with that in adjacent non-neoplastic bone tissues and the osteoblastic cell line. In functional assays, miR-15a inhibited cell proliferation, migration and invasion in U2OS and MG-63 cells. Meanwhile, bioinformatic analysis combined with experimental confirmation demonstrated that tumor necrosis factor; α-induced protein 1 (TNFAIP1) gene is a potential target of miR-15a and can be directly regulated by miR-15a. Down-regulation of TNFAIP1 induced effects on osteosarcoma cell lines similar to those induced by miR-15a. Taken together, these data suggest that miR-15a may act as a tumor suppressor, which is commonly down-regulated in both osteosarcoma tissues and cells. TNFAIP1 plays an important role in mediating miR-15a dependent biological functions in osteosarcoma. Reintroduction of miR-15a may be a novel therapeutic strategy by down-regulating TNFAIP1 expression.