miR-490-3p通过抑制TGFβR1基因表达进而抑制结肠癌细胞的转移和侵袭

目的：研究miR-490-3p在结肠癌细胞(colorectal cancer cell,CRC)转移中的表现和生物功能,及其调控作用机制。方法：通过荧光定量PCR测定miR-490-3p在CRC细胞系的表达水平。细胞转染miR-490-3p以及shmiR-490-3p,观察miR-490-3p的过表达或基因沉默对结肠癌细胞的转移能力是否有影响。miR-490-3p的分子靶标由双荧光素酶报告基因分析和免疫印迹技术进行实验认定。通过划痕实验,Transwell小室基质渗透实验对细胞迁移和侵袭能力进行鉴定。结果：miR-490-3p在CRC细胞系中显著低表达(P<0.05,P<0.01,P<0.001,n>3)。过表达miR-490-3p显著降低CRC细胞株的细胞迁移和侵袭能力(P<0.01,n>3)。miR-490-3p的基因沉默显著增加CRC细胞株的细胞迁移和侵袭能力(P<0.01,n>3)。结肠癌细胞细胞系中过表达miR-490-3p显著降低TGFβR1的基因表达(P<0.001,n>3),miR-490-3p基因沉默显著上调TGFβR1的基因表达(P<0.001,n>3)。过表达miR-490-3p抑制TGFβR1的萤光素酶活性(P<0.001,n>3),miR-490-3p基因沉默促进TGFβR1的萤光素酶活性(P<0.001,n>3)。TGFβR1基因沉默减弱shmiR-490介导的细胞迁移和侵袭促进效应(P<0.01, n>3)。结论：miR-490-3p通过抑制TGFβR1的基因表达从而抑制CRC细胞的转移。     

LncRNA TUG1靶向miR-141-3p影响甲状腺癌细胞的增殖和侵袭

目的 检测LncRNA TUG1在甲状腺癌细胞中的表达,初步探讨LncRNA TUG1对甲状腺癌细胞增殖和侵袭的影响及机制。方法 采用qRT-PCR法检测甲状腺癌B-CPAP、TPC-1细胞株中LncRNA TUG1和miR-141-3p的表达水平,应用生物信息学预测lncRNA TUG1和miR-141-3p的靶向关系,双荧光素酶报告基因实验进行验证。将lncRNA TUG1干扰序列转染TPC-1细胞,应用qRT-PCR法检测lncRNA TUG1和miR-141-3p的表达,MTT法检测各组TPC-1细胞增殖能力,Transwell实验检测各组TPC-1细胞的侵袭能力。结果 与正常甲状腺上皮细胞相比,BCPAP、TPC-1中LncRNA TUG1表达水平明显升高（P<0.05）,miR-141-3p表达水平明显下降（P<0.05）,生物信息学分析及双荧光素酶实验结果显示,lncRNA TUG1可以靶向调控miR-141-3p。转染TUG1干扰序列能够降低TPC-1细胞lncRNA TUG1的表达,上调miR-141-3p的表达（P<0.05）;沉默lncRNA T...     

miR-144-3p对前列腺癌PC3细胞增殖和侵袭的影响及可能机制

目的 探讨miR-144-3p对前列腺癌PC3细胞增殖和侵袭的影响及可能机制.方法 利用脂质体介导的方法进行转染,将前列腺癌PC3细胞分为空白组(未进行转染)、miR-144-3p阴性对照组(转染无义序列)和miR-144-3p模拟物组(转染miR-144-3p模拟物),采用实时定量PCR验证转染效率,Western ...     

miR-483-3p靶向SMAD4对肾上腺皮质癌NCI-H295R细胞增殖和侵袭的影响

目的 探讨miR-483-3p对肾上腺皮质癌NCI-H295R细胞增殖和侵袭的影响及机制.方法 miR-483-3p模拟物和抑制物分别转染肾上腺皮质癌NCI-H295R细胞,设置空白组(未进行转染)、miR-483-3p阴性对照组(转染无义序列)、miR-483-3p模拟物转染组(转染miR-483-3p模拟物)、mi...     

lncRNA PURPL通过调控miR-367-3p/MTA3轴抑制肾癌细胞的增殖和侵袭

目的 观察lncRNA PURPL在肾癌组织中的表达,探讨PURPL/miR-367-3p/MTA3分子轴对肾癌细胞增殖和侵袭的影响及可能的机制.方法 采用qRT-PCR检测PURPL在肾癌组织和不同肾癌细胞系中的表达.选择PURPL表达最低的肾癌细胞系,分别感染阴性对照慢病毒(对照组)和PURPL慢病毒(PURPL组...     

miR-129-5p靶向KLK7对卵巢癌SKOV3细胞增殖和侵袭的影响及分子机制

目的 探讨高表达miR-129-5p对卵巢癌SKOV3细胞增殖和侵袭的影响及分子机制。方法 采用实时荧光定量PCR方法检测卵巢癌SKOV3细胞与人正常卵巢上皮细胞株中miR-129-5p的表达。在卵巢癌SKOV3细胞中瞬时转染miR-129-5p模拟物及无意义序列,采用CCK8法及Transwell实验检测各组细胞的增殖能力和侵袭能力;实时荧光定量PCR检测miR-129-5p和KLK7的相对表达水平;Western blot检测各组细胞中KLK7蛋白表达;Targetscan 7.2和双荧光素酶实验分析miR-129-5p与KLK7的靶向关系。结果 miR-129-5p在卵巢癌SKOV3细胞中的表达低于人正常卵巢上皮细胞(P<0.05),过表达miR-129-5p降低SKOV3细胞中KLK7 mRNA和蛋白的表达,Targetscan 7.2网站分析及双荧光素酶实验证实KLK7为miR-129-5p的靶基因,过表达miR-129-5p抑制SKOV3细胞增殖和侵袭能力(P<0.05)。结论 过表达miR-129-5p可通过靶向调控KLK7抑制卵巢癌SKOV3细胞增殖和侵袭。     

LncRNA TMPO-AS1通过调节miR-204-3p影响肺癌细胞的增殖、凋亡和侵袭

目的:探究长链非编码RNA(lncRNA)TMPO-AS1通过调节miR-204-3p的表达对肺癌细胞增殖、凋亡和侵袭的影响.方法:实时荧光定量PCR(qPCR)检测人肺癌细胞系A549、H1975、H1299和H1650和人正常支气管上皮细胞系HBE中TMPO-AS1的表达水平.采用脂质体转染技术沉默A549细胞中T...     

LncRNA ZFAS1作为ceRNA吸附miR-541-3p调控胃癌侵袭转移北大核心CSCD

目的:本研究旨在探讨长链非编码RNA锌指蛋白反义链1(ZFAS1)作为ceRNA吸附miR-541-3p对胃癌侵袭转移的影响。方法:qRT-PCR对癌旁组织、胃癌组织以及人胃黏膜上皮细胞株、胃癌细胞株中ZFAS1和miR-541-3p的表达进行检测。验证ZFAS1和miR-541-3p的之间的靶向关系。取ZFAS1表达最高的细胞用于后续试验并转染miR-541-3p inhibitor、ZFAS1-shRNA等。MTT检测细胞增殖,Western blot检测侵袭转移相关因子的表达,Transwell测定细胞侵袭,流式细胞术检测细胞凋亡。结果:癌组织中ZFAS1表达增多而miR-541-3p表达减少,ZFAS1能够靶向抑制miR-541的表达(均P<0.05)。抑制ZFAS1表达能够抑制胃癌细胞增殖、侵袭以及侵袭转移相关因子的表达,诱导细胞凋亡,而敲除miR-541-3p后效果则相反(均P<0.05)。ZFAS1-shRNA对胃癌细胞的作用能够被miR-541-3p inhibitor挽救。结论:ZFAS1能够作为ceRNA吸附miR-541-3p,从而抑制胃癌细胞增殖和侵袭转移,促进细胞凋亡。     

微小RNA-140-3p靶向细胞分裂周期相关蛋白8抑制肺腺癌细胞侵袭转移的生物信息学分析及验证

目的 在生物信息学基础上探讨微小RNA(miR)-140-3p靶向细胞分裂周期相关蛋白8(CDCA8)抑制肺腺癌细胞的侵袭和转移.方法 通过GEO数据库中的GEO2R分析肺腺癌芯片数据中差异表达的miRNA.TargetScanHuman7.2和 miRWalk 数据库查找 miR-140-3p 的靶基因.Cytosc...     

miR-2116-3p在人增生性瘢痕中的表达及作用

背景：目前多项研究证实了miRNA影响增生性瘢痕的发生发展,Ⅰ型胶原与增生性瘢痕成纤维细胞密切相关,猜测miR-2116-3p也有可能与增生性瘢痕的发生发展有关系。目的：探讨miR-2116-3p在人增生性瘢痕中的表达及作用。方法：收集新疆医科大学第一附属医院6例患者的增生性瘢痕组织与6例患者重睑术后正常皮肤组织,采用qRT-PCR法检测miR-2116-3p与Ⅰ型胶原的表达。取增生性瘢痕组织,原代培养第3-6代成纤维细胞,分为阴性对照组、miR-2116-3p模拟物组和miR-2116-3p抑制物组,分别转染对应的序列,用CCK-8法与EdU试剂盒检测细胞增殖活力,划痕实验检测细胞迁移能力,流式细胞术检测细胞凋亡,qRT-PCR法与Western blot法检测Ⅰ型胶原、Ⅲ型胶原和α平滑肌肌动蛋白的基因与蛋白表达,双荧光素酶实验验证其靶向结合。结果与结论：(1)增生性瘢痕组织中miR-2116-3p的mRNA表达量低于正常皮肤组织(P <0.01),Ⅰ型胶原的mRNA表达量低于正常皮肤组织(P <0.01);(2)转染后24,48,72 h,与阴性对照组比较,miR-21...     

MicroRNA-483-3p对人神经胶质瘤细胞的作用

目的:探讨microRNA(miRNA)-483-3p对人神经胶质瘤细胞A172生长和迁移能力的影响及潜在作用机制。方法:实时荧光定量聚合酶链式反应(RT-q PCR)检测人肾胚细胞系HEK-293和不同神经胶质瘤细胞株(A172、U251和SHG44)中miRNA-483-3p的表达水平。转染miRNA-483-3p抑制序列(miRNA-483-3p inhibitor)下调A172细胞中miRNA-483-3p的表达,采用CCK-8法和流式细胞术检测细胞活力和周期分布;Transwell实验检测细胞的迁移;Western blot检测周期相关调控因子及上皮-间充质转化相关蛋白的水平。双萤光素酶报告基因分析法预测及验证其可能的靶基因。结果:miRNA-483-3p在各型神经胶质瘤细胞中高表达。沉默miRNA-483-3p后,A172细胞的活力下降并呈现出明显的周期阻滞,且细胞迁移率也显著降低。同时细胞中cyclin D1、周期蛋白依赖性激酶4、磷酸化视网膜母细胞瘤蛋白、N-cadherin及vimentin的蛋白表达水平均显著降低,E-cadherin和β-catenin的蛋白表达水平显著升高。双萤光素酶报告基因分析显示Smad4是miRNA-483-3p的可能作用靶点,A172细胞共转染miRNA-483-3p inhibitor和Smad4 siRNA可部分逆转miRNA-483-3p介导的细胞增殖及迁移抑制。结论:沉默miRNA-483-3p可通过靶向Smad4抑制神经胶质瘤细胞株A172的生长及迁移。     

miR-363-3p在胃癌组织中的表达及其对细胞系HGC-27增殖、迁移与侵袭功能的影响

目的探讨miR-363-3p在胃癌及癌旁样本中的表达差异,并分析其在胃癌细胞系中的功能。方法使用realtime PCR检测59例胃癌组织及对应癌旁组织中miR-363-3p的表达差异;使用miR-363-3p模拟物(miR-363-3pmimic)实现miRNA在胃癌细胞系HGC-27中的过表达,经增殖实验、划痕实验和Transwell实验,检测miR-363-3p对HGC-27细胞功能的影响。结果 miR-363-3p抑制胃癌细胞系HGC-27细胞增殖(P0.05,P0.01),抑制胃癌细胞系HGC-27细胞迁移(P0.001),miR-363-3p对抑制胃癌细胞系细胞的侵袭(P0.05)。结论 miR-363-3p在胃癌发生中可能起到抑癌作用,并有可能成为对胃癌治疗的一个新靶点。     

微小RNA-138-5p抑制肺癌细胞增殖、迁移和侵袭能力的机制研究

目的:探讨微小RNA-138-5p(miR-138-5p)抑制肺癌细胞增殖、迁移和侵袭能力的相关机制。方法:以肺癌细胞A549和H460作为研究对象,分别转染miR-NC(对照组)或miR-138-5p(实验组);生物信息学技术预测miR-138-5p的靶基因;RT-qPCR检测转染后细胞miR-138-5p、叉头框蛋白C1(FOXC1)mRNA和波形蛋白(vimentin)mRNA的相对表达量;Western blot法检测FOXC1、vimentin、E-cadherin、N-cadherin和β-catenin蛋白表达变化;MTS法和集落形成实验分别检测细胞的增殖能力;划痕愈合实验和Transwell法检测细胞迁移和侵袭能力。结果:miR-138-5p过表达显著降低FOXC1和vimentin的mRNA及蛋白的表达(P0.05),E-cadherin和β-catenin蛋白表达上调,N-cadherin蛋白表达下调,显著抑制肺癌细胞的增殖、迁移和侵袭能力(P0.05)。结论:miR-138-5p可以通过靶向干扰FOXC1和vimentin的表达抑制肺癌细胞的增殖、迁移和侵袭,可能是肺癌基因治疗的潜在靶点。     

microRNA-182 inhibits the proliferation and invasion of human lung adenocarcinoma cells through its effect on human cortical actin-associated protein

Lung cancer is one of the main causes of cancer death worldwide. The cortactin gene, CTTN, may play a pivotal role in the proliferation and invasion of tumors. A microRNA (miR-182) was cloned and used to study the expression of CTTN and its regulatory effects on the proliferation and invasion of the lung cancer cell line, A549. Cortactin protein and CTTN mRNA expression decreased in A549 cells that were transfected with the miR-182 expression plasmid. A cell proliferation assay indicated that miR-182 expression affected cell cycle regulation and suppressed proliferation of lung cancer cells in vitro. In addition, xenograft experiments confirmed the suppression of tumor growth in vivo, which was due to the promotion of apoptosis. In conclusion, endogenous mature miR-182 expression may have an important role in the pathogenesis of lung cancer through its interference with the target gene CTTN by epigenetic modification.     

miR-655-3p靶向KIF20A抑制人肺腺癌细胞系A549迁移及侵袭

目的 探讨miR-655-3p靶向驱动蛋白家族成员20A(KIF20A)对肺腺癌细胞增殖、迁移及侵袭的影响.方法 RT-qPCR检测人肺腺癌细胞系H460、A549、HCC-2935、H1299细胞中miR-655-3p的表达;取对数增殖期的A549细胞,分为:空白组、miR-NC组、miR-655-3p mimics...     

MicroRNA-338-3p在肿瘤中的研究进展

微小RNA(microRNA,miRNA)是一种广泛分布于身体各个器官的短链非编码RNA.miRNA可广泛调控基因,通过基因调控控制细胞的生长、分化、增殖以及凋亡等,在正常的细胞生命活动中起着重要作用.随着近年来肿瘤发病率的日益增高,miRNA与肿瘤的关系正成为研究热点,其可作为肿瘤诊断、治疗及预后的标志物.多项研究证实miR-338-3p在多种实质瘤中的表达降低.进一步的研究显示,miR-338-3p可抑制肿瘤基因,通过调控肿瘤基因以及相关信号通路等一系列方式影响肿瘤的进展以及诊断预后.越来越多的证据显示miR-338-3p可作为一种新颖的肿瘤生物标志物,并且有可能成为新的治疗靶点.     

Heparanase promotes human gastric cancer cells migration and invasion by increasing Src and p38 phosphorylation expression

Gastric cancer is one of the most common cancers and it remains difficult to cure, primarily because most cancer stem like cells possess higher capability of invasion and metastasis. Heparanase acts as a master regulator of the aggressive tumor phenotype in part by enhancing expression of proteins and activating signaling molecules. There were less associated with heparanase of molecular biology mechanism in human gastric cancer. We first evaluated the endogenous expression of heparanase in human gastric cancer cell lines and found Heparanase expression higher in SGC-7901 than MGC-803. Using the technology of RNAi in SGC-7901 cells down regulated heparanase gene, and reduced SGC-7901 cells migration and invasion. On the other hand, recombinant heparanase protein added in MGC-803 cells enhanced MGC-803 cell migration and invasion. The elevated cell migration and invasion were impaired by treatment of Src inhibitor pp2 or p38 inhibitor SB 203580. We further found that Stable knockdown of heparanase in SGC-7901 cells decreased phosphorylation of Src and p38. The phosphorylation of p38 was inhibited in response to pp2 treatment while the addition of SB 203580 to SGC-7901 cells did not change phosphorylation of Src. These data suggest that heparanase facilitates invasion and migration of human gastric cancer cells probably through elevating phosphorylation of Src and p38.     

MiR-124-3p inhibits the migration and invasion of Gastric cancer by targeting ITGB3

BACKGROUNDGastric cancer is one of the major malignant tumors in the world. Integrins expressed in cancer cells can promote tumor progression and migration. MiRNAs can inhibit the expression of target genes by directly binding to their mRNAs and can affect various important biological processes. The aim of this study was to investigate the role of miR-124- 3p and ITGB3 in gastric cancer.METHODSRT-PCR and western blot are used to detect the expression of miR-124-3p, ITGB3 and integrin β3 in gastric cancer tissues and cells. The wound healing, CCK-8 assay, transwell migration and invasion assay were performed to determine the cell proliferation, migration and invasion. What’s more, bioinformatics prediction and luciferase assay was conducted to demonstrated the binding efficiency between miR-124-3p and ITGB3.RESULTSWe verified that ITGB3 and miR-124-3p changes the migration and invasion of gastric cancer cells in vitro. The overexpression or silencing of miR-124-3p inhibited or promoted the proliferation, migration and invasion of both selected gastric cancer cells, and ITGB3 is just the reverse. Meanwhile, we validated that ITGB3 is the target of miR-124-3p by bioinformatics prediction and luciferase assay. Lastly, the expression of ITGB3 in 40 pairs of gastric cancer tissues were significantly higher than that in the adjacent normal tissues, while the expression level of miR-124-3p was significantly decreased in cancer tissues.CONCLUSIONSmiR-124-3p inhibits the migration and invasion of Gastric cancer by targeting ITGB3 in gastric cancer cells. Our results suggested that miR-124-3p and ITGB3 may reasonably serve as a promising therapeutic target.