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。     

LncRNA PVT1抑制miR-497-5p表达调控子宫内膜癌细胞增殖和迁移侵袭的分子机制研究

目的:研究长链非编码RNA PVT1和miR-497-5p在子宫内膜癌组织中的表达以及其对癌细胞增殖、迁移和侵袭的影响,并探讨其机制。方法:运用实时荧光定量法测定子宫内膜癌组织、癌旁正常组织及子宫内膜癌细胞HEC-1-B中PVT1和miR-497-5p的表达。用脂质体法将siRNA-PVT1和miR-497-5p mimic转染至子宫内膜癌细胞HEC-1-B;MTT法、Transwell法检测HEC-1-B细胞的增殖、迁移和侵袭。Targetscan在线分析网站预测和双荧光素酶报告基因实验验证LncRNA PVT1和miR-497-5p的靶向关系。将siRNA-PVT1和miR-497-5p inhibitor共转染至HEC-1-B细胞,MTT法、Transwell法检测HEC-1-B细胞的增殖、迁移和侵袭。结果:与癌旁正常组织相比,子宫内膜癌组织中PVT1表达显著上调,miR-497-5p表达显著下调（P<0.05）。沉默PVT1、过表达miR-497-5p均可抑制HEC-1-B细胞的增殖、迁移和侵袭（P<0.05）。PVT1靶向miR-497-5p。抑制miR-497-5p的表达可逆转沉默PVT1对HEC-1-B细胞增殖、迁移和侵袭的抑制作用。结论:沉默PVT1可抑制子宫内膜癌细胞增殖、迁移和侵袭,其机制可能与PVT1靶向miR-497-5p有关,将可为子宫内膜癌的靶向治疗提供新靶点。     

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。     

miR-513a-5p通过靶向调控PAK1抑制子宫颈癌细胞增殖和侵袭

目的 研究miR-513a-5p通过靶向调控p21活化激酶1(p21-activated kinase 1,PAK1)抑制子宫颈癌细胞增殖和侵袭的机制。方法 通过分析高通量基因表达（Gene Expression Omnibus,GEO）数据库GSE145372数据集的数据确认miR-513a-5p在子宫颈癌组织和正常子宫颈组织中的表达差异。采用实时定量聚合酶链式反应（real-time quantitative polymerase chain reaction,RT-qPCR）检测miR-513a-5p和PAK1在子宫颈癌细胞株（HT3、C-33A和HeLa）和人子宫颈内膜上皮细胞株（End1/E6E7）中的表达情况。在miR-513a-5p表达最低的细胞株HT3和C-33A中分别转染miR-513a-5p mimics(miR-513a-5p mimics组)和NC mimics(NC mimics组)。在HT3和C-33A细胞中分别转染pcDNA3.1-PAK1和空载pcDNA3.1-NC用于后续功能拯救实验。采用EdU实验和transwell实验分别检测miR-513a-5p...     

沉默circZFR通过调控miR-497-5p表达抑制脊索瘤细胞的增殖、迁移及侵袭

目的:研究环状RNA（circular RNAs,circRNAs）circZFR是否通过调控微小RNA（microRNA,miRNA/miR）-497-5p的表达影响脊索瘤细胞增殖、迁移及侵袭。方法:实时荧光定量PCR（quantitative real-time polymerase chain reaction,qRT-PCR）检测脊索瘤组织中circZFR和miR-497-5p的表达。在脊索瘤U-CH1细胞中转染si-circZFR或miR-497-5p。噻唑蓝（methyl thiazolyl tetrazolium,MTT）检测细胞增殖,Transwell小室法检测细胞迁移及侵袭情况,蛋白质印迹法（Western blot）检测细胞周期蛋白D1（CyclinD1）、p21、基质金属蛋白酶-2（matrix metalloprotease-2,MMP-2）、基质金属蛋白酶-9（matrix metalloprotease-9,MMP-9）蛋白表达,StarBase预测工具和双荧光素酶报告实验分析circZFR与miR-497-5p的靶向结合。si-circZFR和anti-miR-497-5p共转染,观察下调miR-497-5p表达对沉默circZFR表达诱导的U-CH1细胞增殖、迁移及侵袭的作用。结果:与髓核组织比较,脊索瘤组织中的circZFR表达量明显增加,miR-497-5p表达量显著减少（P＜0.05）。沉默circZFR表达或过表达miR-497-5p明显降低U-CH1细胞48 h、72 h的细胞活性、迁移细胞数、侵袭细胞数、CyclinD1、MMP-2、MMP-9蛋白表达量,显著提高p21蛋白水平（P＜0.05）。circZFR靶向调控miR-497-5p的表达。下调miR-497-5p表达逆转了沉默circZFR表达对脊索瘤U-CH1细胞增殖、迁移、侵袭、CyclinD1、MMP-2及MMP-9蛋白表达的抑制作用,和对p21蛋白表达的促进作用。结论:沉默circZFR表达可以抑制脊索瘤细胞的增殖、迁移及侵袭,其作用机制与靶向调控miR-497-5p的表达有关。     

MiR-129-5p靶向HMGB1抑制骨肉瘤细胞增殖和迁移

目的 探讨miR-129-5p对骨肉瘤(OS)细胞增殖和迁移的影响以及对HMGB1的调控作用.方法 RT-PCR和Western blot法分别检测骨肉瘤细胞株MG-63、Saos-2和成骨细胞hFOB1.19中miR-129-5p和HMGB1的表达.生物信息学预测miR-129-5p与HMGB1基因是否存在结合位点,...     

miR-324-5p通过靶向调控DAPK1降低胰腺癌细胞的放射敏感性

目的:探讨死亡相关蛋白激酶1（death-associated protein kinase 1,DAPK1）在胰腺癌（pancreatic cancer,PaC）细胞放射敏感性中的作用,验证miR-324-5p通过靶向调控DAPK1影响胰腺癌细胞放射敏感性的机制。方法:通过生物信息学预测靶向DAPK1的miRNAs,并利用双荧光素酶报告基因检测miR-324-5p对DAPK1的调控作用。在PANC-1和MIA PaCa-2细胞中过表达miR-324-5p和DAPK1或抑制miR-324-5p后,对各细胞株进行放射诱导,检测细胞增殖和凋亡情况,以及凋亡相关分子的表达情况。结果:GEO数据集结果显示,胰腺癌组织中miR-324-5p的表达水平高于正常组织。与正常胰腺导管上皮细胞系（HPDE6-C7）相比,胰腺癌细胞系(Capan-1、Bxpc-3、PANC-1和MIA PaCa-2)中miR-324-5p表达水平更高（P均<0.001）。双荧光素报告基因检测结果表明,miR-324-5p靶向DAPK1的3' UTR,并且可下调DAPK1的表达。细胞实验结果证实,过表达miR-324-5p通过靶向调控DAPK1降低放射诱导的细胞凋亡和DNA的损伤,进而降低了胰腺癌细胞的放射敏感性。结论:miR-324-5p通过负调控DAPK1降低胰腺癌细胞对放射的敏感性,从而影响DNA修复和细胞凋亡。miR-324-5p/DAPK1途径可能为胰腺癌的靶向治疗提供了潜在的治疗靶点。     

miR-497-5p通过HMGA2调控食管鳞癌细胞迁移和侵袭

目的:探讨miR-497-5p通过高迁移率族蛋白A2(HMGA2)对食管鳞癌细胞(ESCC)迁移和侵袭能力的影响。方法:利用UALCAN、GEPIA数据库分析miR-497-5p和HMGA2 mRNA在食管癌中的表达。通过生物信息学网站starBase分析miR-497-5p和HMGA2在食管癌样本中表达的相关性。通过双荧光素酶报告基因实验检测miR-497-5p对HMGA2基因的调控。采用细胞划痕实验和Transwell实验分析miR-497-5p通过HMGA2对ESCC细胞迁移和侵袭能力的影响。结果:miR-497-5p在食管癌中的表达明显低于其对照样本(P<0.05),HMGA2 mRNA在食管癌中的表达明显高于其对照样本(P<0.05)。miR-497-5p和HMGA2的表达成负相关,miR-497-5p对靶基因HMGA2具有直接调控作用。miR-497-5p mimic组细胞的迁移和侵袭能力明显低于阴性对照(NC)组(P<0.001);而miR-497-5p mimic+HMGA2组细胞的迁移和侵袭能力与miR-497-5p mimic组相比明显上升(P&l...     

lncRNA FOXD2-AS1通过靶向miR-506-5p调控宫颈癌细胞增殖与凋亡

[摘要] 目的: 探讨长链非编码RNA（lncRNA）FOXD2-AS1 是否通过靶向miR-506-5p 调控宫颈癌细胞增殖和凋亡。方法:体外培养人宫颈癌细胞系HeLa、Siha、Caski 与正常宫颈细胞株Ect1/E6E7,用qPCR检测细胞中FOXD2-AS1 和miR-506-5p 的表达水平。用脂质体转染技术分别构建抑制FOXD2-AS1 表达和过表达miR-506-5p 的宫颈癌细胞,用MTT实验和流式细胞术检测细胞的增殖和凋亡情况,WB实验检测细胞中增殖相关蛋白CyclinD1、p21 和p27 及凋亡相关蛋白Bcl-2、BAX和cleaved-capase-3的表达。用双荧光素酶报告基因实验验证FOXD2-AS1 是否靶向miR-506-5p,并分析同时抑制FOXD2-AS1 和miR-506-5p 表达对宫颈癌细胞增殖与凋亡的影响。结果：与Ect1/E6E7 细胞比较,宫颈癌HeLa、Siha 和Caski 细胞中FOXD2-AS1 表达水平显著升高,miR-506-5p 表达水平降低（均P<0.01）。抑制FOXD2-AS1 表达可显著抑制宫颈癌细胞中CyclinD1 蛋白和Bcl-2 蛋白表达,并促进p21、p27 和BAX、cleaved-capase-3 蛋白的表达,抑制细胞增殖并促进细胞凋亡（均P<0.01）。过表达miR-506-5p 可显著抑制宫颈癌细胞中CyclinD1 和Bcl-2 蛋白表达,促进p21 和BAX蛋白表达,抑制细胞增殖并促进细胞凋亡（均P<0.01）。双荧光素酶报告基因实验证实宫颈癌细胞中FOXD2-AS1 靶向负调控miR-506-5p 的表达（P<0.01）。抑制miR-506-5p 表达逆转了抑制FOXD2-AS1 表达对宫颈癌细胞增殖和凋亡的作用（P<0.01）。结论: FOXD2-AS1 通过靶向miR-506-5p 的表达调控宫颈癌细胞的增殖和凋亡。     

miR-497 and miR-34a retard lung cancer growth by co-inhibiting cyclin E1 (CCNE1)

Cyclin E1, encoded by the CCNE1 gene, promotes G1/S transition, chromosome instability, and oncogenesis. Here, we show that miR-497 and miR-34a target the 3′-UTR of CCNE1. miR-497 and miR-34a are downregulated in cancer cells and their ectopic expression inhibited cell proliferation and colony formation in vitro, and inhibited tumor growth in a xenograft model. The effect of simultaneous overexpression of miR-497 and miR-34a on the inhibition of cell proliferation, colony formation, and tumor growth, and the downregulation of cyclin E1 was stronger than the effect of each miRNA alone. The synergistic actions of miR-497 and miR-34a partly correlated with cyclin E1 levels. When cells stably expressing CCNE1 were transfected with the Hi-miR-497/34a plasmid, there was no effect on colony formation, compared with that of cells transfected with either Hi-miR497 or Hi-miR34a. These results indicate cyclin E1 is downregulated by both miR-497 and miR-34a, which synergistically retard the growth of human lung cancer cells.     

Upregulation of miR-324-5p Inhibits Proliferation and Invasion of Colorectal Cancer Cells by Targeting ELAVL1

Colorectal cancer (CRC) is a common clinical cancer that remains incurable in most cases. miRNAs are reported to play a part in the development of various tumors. In the present study, we found that miR-324-5p was downregulated in CRC cells, while ELAV (embryonic lethal, abnormal vision, Drosophila)-like protein 1 (ELAVL1) showed a higher expression. miR-324-5p transfection significantly inhibited the proliferation as well as invasion in both SW620 and SW480 cells. miR-324-5p mimic transfection markedly decreased the expression of ELAVL1. Luciferase reporter gene assay confirmed that ELAVL1 is a direct target of miR- 324-5p. Furthermore, cancer invasion factors uPA, uPAR, and MMP-9 were found to drop significantly in miR-324-5p-transfected groups. To conclude, our findings indicate that miR-324-5p may play a suppressive role in colorectal cell viability and invasion, at least in part, through directly targeting ELAVL1. Therefore, miR-234-5p might function as a promising candidate for CRC treatment and deserves deeper research.     

miR-363-3p Inhibits Osteosarcoma Cell Proliferation and Invasion via Targeting SOX4

miR-363-3p has been shown to suppress tumor growth and metastasis in various human cancers. However, the function of miR-363-3p in osteosarcoma (OS) has not been determined. In our study, we found that the expression of miR-363-3p was significantly downregulated in OS tissues compared with adjacent normal tissues. miR-363-3p expression was associated with the poor overall survival rate of OS patients. Moreover, we found that overexpression of miR-363-3p markedly inhibited the proliferation, migration, and invasion of U2OS and MG63 cells. Moreover, we found that SOX4 was a direct target of miR-363-3p in OS cells. Overexpression of miR-363-3p significantly inhibited the expression of SOX4. Expression levels of miR-363-3p and SOX4 were negatively correlated in OS tissues. Finally, we found that restoration of SOX4 attenuated the suppressive effects of miR-363-3p on the proliferation, migration, and invasion of U2OS and MG63 cells. Therefore, our findings demonstrated that miR-363-3p served as a tumor suppressor in OS tissues by targeting SOX4.     

miR-1266-5p and miR-185-5p Promote Cell Apoptosis in Human Prostate Cancer Cell Lines

Objective: Small non-coding RNA molecules are dysregulated in prostate cancer (PCa). In our previous study,downregulation of miR-1266 and miR-185 was demonstrated in PCa tissues and cell lines. The aim of the presentstudy was to investigate whether miR-1266 and miR-185 are involved in the regulation of B-cell lymphoma (BCL) 2and BCL2L1, respectively, and whether transfection of PCa cell lines with miR-1266 and miR-185 mimics can altertumorigenic phenotypes. Methods: In order to investigate the regulation of BCL2 and BCL2L1 mRNA levels bymiR-1266 and miR-185, respectively, a luciferase reporter assay was used. Real-time PCR was also used to analyzechanges in the levels of BCL2 and BCL2L1 mRNAs in PCa cell lines following transfection with synthetic miR-1266and miR-185. Cell apoptosis was determined by Annexin V protein expression analysis via flow cytometry. In additionto the MTT assay, a cell proliferation assay was performed. Result: A luciferase assay confirmed that the BCL2 andBCL2L1 genes may be targeted by miR-1266 and miR-185, respectively, through binding to their 3′UTR regions.Transfection of PC3 and DU145 cells with miR-1266 and miR-185 induced apoptosis and reduced proliferation, whichalso revealed an inverse correlation with BCL2 and BCL2L1 gene expression in the treated cells. Conclusion: Ourdata suggests that miR-1266 and miR-185 may be novel candidates for further research in PCa treatment through theanti-apoptotic pathway.     

The Effects of Combination of Mimic miR-155-5p and Antagonist miR-324-5p Encapsulated Chitosan in Ovarian Cancer SKOV3

Objective: Ovarian cancer is a malignant tumor that attacks reproductive organs of women. MicroRNA is known to have an involvement in the prognosis of ovarian cancer. One of them is miR-155-5p which is down regulated and miR-324-5p which is up regulated. Chitosan is used as microRNA delivery system. The aims of this study is to find out the effects of combination microRNA encapsulated chitosan in cell line SKOV3. Methods: Cell line SKOV3 obtained from Stem Cell and Cancer Institute (Kalbe). Mimic miR-155-5p and Antagonist miR-324-5p formulated with chitosan. Total RNA was extracted from nine samples (three as control and six as treatment), and prepared for cDNA synthesis. Expression of RNA and mRNA target was measured using q-PCR Biorad CFX96 C.100 and Gen Ex 7 software. Statistics analysis was measured using SPSS 16.0. Results: The administration of combination microRNA encapsulated with chitosan affect the expression of miR-155-5p and miR-324-5p endogen (p <0.05). The expression of mRNA target HIF1α and GLI1 was down regulated after treatment. The correlation between expression of microRNA and mRNA target was strongly (p <0.05). Conclusion: This study successfully presented effects of combination of mimic miR-155-5p and antagonist miR-324-5p encapsulated chitosan which be considered as a potential therapy targets for ovarium cancer.