lncRNA MNX1-AS1 Promotes Glioblastoma Progression Through Inhibition of miR-4443

Long noncoding RNAs (lncRNAs) have been acknowledged as important regulators in various human cancers. lncRNA MNX1-AS1 has been shown to be an oncogene in epithelial ovarian cancer. However, the function of MNX1-AS1 in glioblastoma (GBM) remains largely unknown. Here we found that the expression of MNX1-AS1 was significantly upregulated in GBM tissues and cell lines. Knockdown of MNX1-AS1 significantly inhibited the proliferation, migration, and invasion of GBM cells. In terms of mechanism, we found that MNX1-AS1 could bind to miR-4443 in GBM cells. Overexpression of miR-4443 significantly inhibited the expression of MNX1-AS1 and vice versa. Moreover, there was an inverse correlation between the expression levels of MNX1-AS1 and miR-4443 in GBM tissues. We found that overexpression of miR-4443 inhibited the proliferation, migration, and invasion of GBM cells. We also showed that inhibition of miR-4443 reversed the effects of MNX1-AS1 knockdown on GBM cell proliferation, migration, and invasion. Taken together, we found that MNX1-AS1 promoted the proliferation, migration, and invasion of GBM cells through inhibiting miR-4443.     

The Interaction Between lncRNA SNHG1 and miR-140 in Regulating Growth and Tumorigenesis via the TLR4/NF-kB Pathway in Cholangiocarcinoma

Cholangiocarcinoma (CCA) is the second most common primary hepatobiliary carcinoma. The long noncoding RNA (lncRNA) small nucleolar RNA host gene 1 (SNHG1) has been reported to contribute to the progression of multiple cancers. Nonetheless, the functions and hidden mechanism of SNHG1 remain unclear in CCA. In this study, the SNHG1 levels were boosted in CCA cell lines, and knockdown of SNHG1 repressed CCA cell proliferation and invasion in vitro. The data also demonstrated that miR-140 could act as a target of SNHG1 in CCA and inhibited CCA cell proliferation and invasion, whereas the inhibition effects were relieved by overexpression of SNHG1. In addition, Toll-like receptor 4 (TLR4), an NF- B-activating signal, was identified to be a target of miR-140. SNHG1, as a competing endogenous RNA (ceRNA) for miR-140, enhanced TLR4 expression and activated NF- B signaling, thereby regulating growth and tumorigenesis in CCA. Animal experiments further confirmed this conclusion. Collectively, these findings not only uncovered a key role of SNHG1/miR-140/TLR4/NF- B signaling axis in CCA tumorigenesis and progression but also denoted the probable utilization of SNHG1 as a therapeutic target for CCA.     

miR-186 Regulates Glycolysis through Glut1 During the Formation of Cancer-associated Fibroblasts

Emerging evidence has suggested that glycolysis is enhanced in cancer-associated fibroblasts (CAF), andmiR-186 is downregulated during the CAF formation. However, it is not clear whether miR-186 is involved in theregulation of glycolysis and what the role of miR-186 plays during the CAF formation. In this study, quantitativePCR analysises show miR-186 is downregulated during the CAF formation. Moreover, miR-186 targets the 3’UTR of Glut1, and its overexpression results in the degradation of Glut1 mRNA, which eventually reduces thelevel of Glut1 protein. On the other hand, knockdown of miR-186 increased the expression of Glut1. Both timecourse and dose response experiments also demonstrated that the protein and mRNA levels of Glut1 increaseduring CAF formation, according to Western blot and quantitative PCR analyses, respectively. Most importantly,besides the regulation on cell cycle progression, miR-186 regulates glucose uptake and lactate production whichis mediated by Glut1. These observations suggest that miR-186 plays important roles in glycolysis regulation aswell as cell cycle checkpoint activation.     

miR-186通过下调垂体瘤转化基因1的表达抑制骨肉瘤细胞的增殖侵袭并诱导其凋亡

目的:探讨miR-186对骨肉瘤细胞增殖、凋亡及侵袭能力的影响,并探讨其可能机制.方法:实时荧光定量PCR检测骨肉瘤细胞HOS、U2-OS、Saos-2及成骨细胞NHOst中miR-186表达,并运用人工合成的miR-186模拟片段及对照scramble mimic转染至人骨肉瘤HOS及U2-OS细胞内,运用实时荧光定量PCR检测转染后细胞中miR-186的表达水平;分别运用CCK-8法、流式细胞检测技术及Transwall体外侵袭实验检测过表达miR-186对HOS及U2-OS细胞增殖、凋亡及侵袭能力的影响;运用Westem blotting及实时荧光定量PCR检测miR-186过表达对细胞中垂体瘤转化基因1(pituitary tumor transforming gene 1,PTTG1)的蛋白及mRNA表达水平的影响.结果:骨肉瘤细胞中miR-186呈现低表达;转染人工合成的miR-186片段可上调HOS及U2-OS细胞中miR-186的表达;miR-186过表达组细胞的增殖能力较scramble组明显下降(P＜0.01),转染组HOS[(16.9±2.1)％ vs(10.4±1.6)％,P＜0.05]及U2-OS[(22.6±2.9)％ vs (14.1±2.2)％,P＜0.05]细胞的凋亡比例明显高于scramble组,转染组HOS及U2-OS细胞的穿膜数明显低于scramble组(P＜0.01);转染组细胞中PTTG1的蛋白及mRNA表达水平均明显下降(P＜0.01),而scramble组无明显变化(P＞0.05);结论:miR-186能够抑制骨肉瘤细胞的增殖及侵袭并促进细胞凋亡,其作用机制可能与抑制PTTG1表达相关.     

miR-1269a调控HOXD10基因抑制胆管癌细胞侵袭的作用及其机制

目的 探讨miR-1269a对HOXD10基因的调控作用及对胆管癌细胞侵袭能力的影响。方法 预测并筛选出调控HOXD10基因的miR-1269a作为研究对象;转染miR-1269a模拟物（mimic）及抑制剂（inhibitor）至胆管癌细胞系后检测 HOXD10mRNA及蛋白的表达变化,并观察miR-1269a对胆管癌细胞侵袭能力的影响。双荧光素酶报告基因实验验证miR-1269a与HOXD10之间的靶向作用关系。结果 miR-1269a在胆管癌组织中表达显著高于癌旁组织（P=0.0023）;miR-1269a模拟物可显著降低胆管癌细胞中HOXD10 mRNA（Mz-CHA-1: P=0.0025; RBE: P=0.0038）及蛋白表达水平,且细胞的侵袭能力较对照组显著增强（Mz-CHA-1: P=0.004; RBE: P=0.004）。miR-1269a抑制剂转染则出现相反的结果（QBC939: P=0.16; HCCC9810: P=0.13）。miR-1269a明显抑制野生型HOXD10-3’UTR 的荧光素酶活性,而对突变型质粒转染细胞的荧光素酶活性无影响。结论 miR-1269a靶向负性调控HOXD10基因,在胆管癌细胞侵袭过程中发挥重要作用。     

The lncRNA FEZF1-AS1 Promotes the Progression of Colorectal Cancer Through Regulating OTX1 and Targeting miR-30a-5p

Long noncoding RNAs (lncRNAs) participate in and regulate the biological process of colorectal cancer (CRC) progression. Our previous research identified differentially expressed lncRNAs in 10 CRC tissues and 10 matched nontumor tissues by next-generation sequencing (NGS). In this study, we identified an lncRNA, FEZF1 antisense RNA 1 (FEZF1-AS1), and further explored its function and mechanism in CRC. We verified that FEZF1-AS1 is highly expressed in CRC tissues and cell lines. Through functional experiments, we found that reduced levels of FEZF1-AS1 significantly suppressed CRC cell migration, invasion, and proliferation and inhibited tumor growth in vivo. Mechanistically, we discovered that reduced levels of the lncRNA FEZF1- AS1 inhibited the activation of epithelial–mesenchymal transition (EMT); the overexpression of orthodenticle homeobox 1 (OTX1) partially rescued the FEZF1-AS1-induced inhibition of protein expression. It indicated that FEZF1-AS1 may play a role in the occurrence and development of CRC by regulating the FEZF1-AS1/ OTX1/EMT pathway. Furthermore, it was reported that FEZF1-AS1 is located in both the nucleus and cytoplasm of HCT116 cells. Dual-luciferase reporter assays verified that FEZF1-AS1 directly binds miR-30a-5p and negatively regulated each other. Further, we showed that 5 -nucleotidase ecto (NT5E) is a direct target of miR-30a-5p, and the inhibition of miR-30a-5p expression partially rescued the inhibitory effect of FEZF1-AS1 on NT5E. Our results indicated that the mechanism by which FEZF1-AS1 positively regulates the expression of NT5E is through sponging miR-30a-5p. Our study demonstrated that lncRNA FEZF1-AS1 is involved in the development of CRC and may serve as a diagnostic and therapeutic target for CRC patients.     

姜黄素通过下调miR-186*促进人肺腺癌细胞 A549/DDP凋亡

背景与目的姜黄素是从姜黄科植物的根茎中提取的一种天然化合物。体内外的临床前研究表明,其具有抗炎、抗氧化、抗肿瘤等多种作用。miR-186*是通过microarray芯片技术发现的在人肺腺癌细胞A549/DDP细胞中高表达的基因。本研究旨在阐明姜黄素是否可通过调控miR-186*的表达而促进A549/DDP细胞的凋亡。方法mi-croarray芯片技术检测经姜黄素、DMSO分别处理后A549/DDP细胞中microRNAs(miRNAs)表达量的变化;实时定量PCR验证从microarray芯片中筛选出来的明显差异表达的miRNAs;最后,流式细胞仪法检测明显差异表达的miR-NAs调控的细胞凋亡,MTT法检测细胞存活率。结果microarray芯片技术结果显示:与DMSO对照组比较,姜黄素处理后miR-186*的表达明显下调。实时定量PCR验证了microarray芯片技术检测的结果。流式细胞仪法检测miR-186*调控的细胞凋亡,MTT法检测细胞存活率:姜黄素处理后,与对照组比较,miR-186*的表达明显下调,从而加速A549/DDP细胞的凋亡,细胞存活率下降;转染miR-186*的mimetics和其对...     

Twist1对人乳腺癌细胞E-cadher in表达及亚细胞定位的影响

  目的   探讨转录调控因子Twist1对人乳腺癌MCF-7细胞E-cadherin表达的影响。   方法   上调人乳腺癌MCF-7细胞中Twist1的表达, 利用RT-PCR, Western blot, 细胞免疫荧光检测分析上调Twist1对E-cadherin表达的影响; 划痕实验分析对细胞侵袭运动能力的影响; 三维培养观察Twist1对MCF-7细胞成血管能力的影响。   结果   MCF-7细胞过表达Twist1抑制细胞E-cadherin的表达, 并且导致其亚细胞定位异常, 细胞迁移运动能力增强, 细胞成血管能力增强。   结论   Twist1能抑制MCF-7细胞E-cadherin的表达, 并增强其侵袭迁移及血管生成能力。      

miR-186 Represses Proliferation,Migration, Invasion,and EMT of Hepatocellular Carcinoma via Directly Targeting CDK6

The present study aimed to investigate the effect of miR-186 on proliferation, migration, invasion, and epithelial–mesenchymal transition (EMT) of hepatocellular carcinoma (HCC). In this work, miR-186 was downregulated in HCC tissues and cells, and low miR-186 level helped predict the occurrence of vascular invasion and poor prognosis in patients with HCC. miR-186 overexpression inhibited cell proliferation and tumor growth in nude mice, repressed migration and invasion abilities, and enhanced apoptosis in HCC cells. miR-186 also retarded progression of EMT. miR-186 directly bound to the 3 -untranslated regions of cyclin-dependent kinase 6 (CDK6) to inhibit its expression. Overexpression of CDK6 markedly reversed inhibitory effects of miR-186 on proliferation, apoptosis, migration, and invasion of HCC cells. Conversely, inhibition of CDK6 exerted synergic effect on the biological functions of miR-186. In conclusion, miR-186 represses proliferation, migration, invasion, and EMT, and induces apoptosis through targeting CDK6 in HCC, which may provide a new therapeutic target for HCC.     

The PDGF-D/miR-106a/Twist1 pathway orchestrates epithelial-mesenchymal transition in gemcitabine resistance hepatoma cells

Emerging evidence demonstrates that platelet-derived growth factor-D (PDGF-D) plays a critical role in epithelial-mesenchymal transition (EMT) and drug resistance in hepatocellular carcinoma (HCC) cells. However, the underlying mechanism has not been fully elucidated. The objective is to explore the molecular mechanism of PDGF-D-mediated EMT in drug resistance HCC cells. To achieve our goal, we used multiple approaches including Western blotting, real-time RT-PCR, wound healing assay, invasion assay, luciferase activity assay, transfection, and immunohistochemistry. We found that PDGF-D is highly expressed in gemcitabine-resistant (GR) HCC cells. Moreover, PDGF-D markedly inhibited miR-106a expression and subsequently upregulated Twist1 expression. Notably, PDGF-D expression was associated with miR-106a and Twist1 in HCC patients. Our findings provide a possible molecular mechanism for understanding GR chemoresistance in HCC cells. Therefore, inactivation of PDGF-D/Twist or activation of miR-106a could be a novel strategy for the treatment of HCC.     

MicroRNA 495 Inhibits Proliferation and Metastasis and Promotes Apoptosis by Targeting Twist1 in Gastric Cancer Cells

Recently, microRNAs (miRNAs) have been reported to participate in multiple biological processes. However, the effects of miR-495 on gastric cancer (GC) remain unclear. The purpose of this study was to explore the functions of miR-495 in GC cell proliferation, metastasis, and apoptosis. SGC-7901 and BGC-823 cell lines were transfected with miR-495 mimic, miR-495 inhibitor, and negative controls (mimic control and inhibitor control). The expressions of miR-495, cell viability, migration, apoptosis, and apoptosis-related factors were examined by qRT-PCR, trypan blue staining, Transwell, flow cytometry, and Western blot, respectively. Simultaneously, key factor expression levels of EMT were detected by qRT-PCR and Western blot. The direct target of miR-495 was confirmed by dual-luciferase assay. Additionally, sh-Twist1, pc-Twist1, and corresponding controls were transfected into SGC-7901 and BGC-823 cells, and the protein levels of EMTassociated factors were detected by Western blot. miR-495 was downregulated in GC cells. miR-495 expression level was effectively overexpressed or suppressed in SGC-7901 and BGC-823 cells. Overexpression of miR-495 significantly decreased cell viability and migration, increased apoptosis, and inhibited the EMT process. Suppression of miR-495 showed contrary results. Twist1 was clarified as a target gene of miR-495, and Twist1 silencing obviously reduced the promoting effect of miR-495 suppression on these biological processes. Twist1 silencing significantly blocked the EMT process in both SGC-7901 and BGC-823 cells. miR-495 inhibited proliferation and metastasis and promoted apoptosis by targeting Twist1 in GC cells. These data indicated that miR-495 might be a novel antitumor factor of GC and provide a new method for the treatment of GC.     

CRNDE affects the malignant biological characteristics of human glioma stem cells by negatively regulating miR-186

The long non-coding RNA Colorectal neoplasia differentially expressed (CRNDE) is a novel gene that activated early in colorectal neoplasia, but it is also up-regulated in many other solid tumors. Herein, the function and underlying mechanism of CRNDE in regulating glioma stem cells (GSCs) were investigated. We found that CRNDE expression was up-regulated while miR-186 expression was down-regulated in GSCs. Overexpression of CRNDE could promote the cellular proliferation, migration, invasion and inhibit the apoptosis in GSCs. Overexpression of miR-186 exerted functions of inhibiting the proliferation, migration and invasion of GSCs and promoting apoptosis. And CRNDE decreased the expression levels of XIAP and PAK7 by binding to miR-186 and negatively regulating it. In addition, miR-186 binded to XIAP and PAK7 3′UTR region, and decrease the expression of them, thus regulating the expression levels of downstream target proteins such as caspase 3, BAD, cyclin D1 and MARK2. The in vivo effect of CRNDE and miR-186 showed that the tumor formation rate was minimum in tumor-bearing nude mice with the knockdown of CRNDE and the overexpression of miR-186. In conclusion, CRNDE played an oncogenic role of GSCs through the negative regulation of miR-186. Both CRNDE and miR-186 could be regarded as potential targets in the glioma therapy.     

miR-374a Inhibitor Enhances Etoposide-Induced Cytotoxicity Against Glioma Cells Through Upregulation of FOXO1

Glioma is a commonly diagnosed brain tumor that shows high mortality rate. Despite the great advancement of cancer therapy in recent years, chemotherapy is still an important approach for treatment of glioma. However, long-term chemotherapy usually causes serious side effects or complications. It is desirable to take strategies to enhance the efficacy of current chemotherapy. In the present study, we observed obvious upregulation of miR-374a in glioma cells. More importantly, we found that knockdown of miR-374a was able to enhance the etoposide-induced cytotoxicity against glioma cells. Mechanically, we demonstrated that FOXO1 was the target of miR-374a in glioma. Treatment with miR-374a inhibitor induced overexpression of FOXO1, and thus promoted the expression of Bim and Noxa. Since Bim and Noxa act as key proapoptotic proteins in mitochondrial apoptosis, miR-374a inhibitor was able to enhance the etoposide-induced apoptosis pathway in glioma.     

Cinobufacin Suppresses Cell Proliferation via miR-494 in BGC-823 Gastric Cancer Cells

Cinobufacin is used clinically to treat patients with many solid malignant tumors. However, the mechanismsunderlying action remain to be detailed. Our study focused on miRNAs involved in cinobufacin inhibition of GCcell proliferation. miRNA microarray analysis and real time PCR identified miR-494 as a significant cinobufacinassociatedmiRNA. In vivo, ectopic expression of miR-494 inhibited the proliferation and induced apoptosis ofBGC-823 cells on CCK-8 and flow cytometry analysis. Further study verified BAG-1 (anti-apoptosis gene) to beatarget of miR-494 by luciferase reporter assay and Western blotting. In summary, our study demonstrated thatcinobufacin may inhibit the proliferation and promote the apoptosis of BGC-823 cells. Cinobufacin-associatedmiR-494 may indirectly be involved in cell proliferation and apoptosis by targeting BAG-1, pointing to use as apotential molecular target of cinobufacin in gastric cancer therapy.     

miR-150 Suppresses Tumor Growth in Melanoma Through Downregulation of MYB

miR-150 has been demonstrated to inhibit tumor progression in various human cancers, including colorectal cancer, ovarian cancer, and thyroid cancer. However, the role of miR-150 in melanoma remains to be determined. In this study, we found that miR-150 was underexpressed in melanoma tissues and cell lines. Through transfection of miR-150 mimics, we found that miR-150 significantly inhibited the proliferation, migration, and invasion of melanoma cells. In mechanism, we found that MYB was a target of miR-150 in melanoma cells. Overexpression of miR-150 significantly inhibited mRNA and protein levels of MYB in melanoma cells. Moreover, there was an inverse correlation between the expression of miR-150 and MYB in melanoma tissues. We also showed that MYB was upregulated in melanoma tissues and cell lines. Through functional experiments, we found that restoration of MYB in miR-150-overexpressed melanoma cells rescued the proliferation, migration, and invasion. Therefore, our findings demonstrated that miR-150 suppressed the proliferation, migration, and invasion of melanoma cell by downregulating MYB.     

Progression of targeted therapy in advanced cholangiocarcinoma

It is necessary to establish an effective therapy to improve the survival of patients with advanced cholangiocarcinoma (CCA). Recently, with the development of pathology research in CCA, a lot of special bio-markers such as EGFR, VEGF, HER2, and MEK et al. could be over expression or mutations in CCA patients. According to their changes, combinations of targeted therapy plus chemotherapy are now recognized as effective therapies for advanced CCA. The aim of this paper is to analyze recent promising studies about targeted therapy alone or combination with each other or with chemotherapies.     

miR-505抑制无脑回致病基因进而抑制肝内胆管细胞癌RBE细胞增殖和侵袭

目的:观察miR-505对肝内胆管细胞癌(intrahepatic cholangiocarcinoma,ICC)细胞的增殖、S期阻滞和侵袭的影响,寻找miR-505在调控过程中的关键靶基因,阐明miR-505在肝内胆管细胞癌中的作用及其工作机制。方法:分析ICC组织内miR-505和LIS1表达变化,采用TargetScan和双荧光素酶报告基因试验证实LIS1是否为miR-505的靶基因。采用CCK8法、流式细胞法和Transwell实验分别检测miR-505和LIS1对RBE细胞增殖、S期细胞比值和侵袭的影响。蛋白质印迹法检测miR-505对MMP-2/p-AKT的影响。结果:miR-505在ICC组织中的表达降低,L1S1是miR-505靶基因,同时相比对照组,ICC组织中LIS1异常高表达。随后体外细胞实验发现转染模拟物能抑制RBE细胞的增殖,S期阻滞和侵袭,而LIS1能逆转这种抑制效果。过表达miR-505可以抑制MMP-2/p-AKT的表达。结论:miR-505表达的失调与ICC密切相关,并能通过直接靶向作用于LIS1抑制MMP-2/p-AKT的表达,从而参与调控ICC细胞的生物学行为,发挥抑制ICC的作用。     

miR-449a Suppresses Tumor Growth,Migration, and Invasion in Non-Small Cell Lung Cancer by Targeting a HMGB1-Mediated NF-kB Signaling Pathway

MicroRNAs (miRNAs) have been reported to be involved in many human cancers and tumor progression. The dysregulation of miR-449a is found in many types of malignancies and is associated with tumor growth, migration, and invasion. However, its expression and function in non-small cell lung cancer (NSCLC) still remains unclear. In our study, miR-449a was found to be downregulated in both NSCLC tissues and cell lines, and low miR-449a expression was obviously associated with tumor differentiation, TMN stage, and poor overall survival (OS). Moreover, we demonstrated that miR-449a could inhibit tumor proliferation, migration, and invasion in NSCLC. We also confirmed that HMGB1 was a direct target gene of miR-449a in NSCLC with dual-luciferase reporter assay, and upregulation of HMGB1 could reverse the miR-449a-induced suppression of growth, migration, and invasion in NSCLC cells. Last, we found that miR-449a suppressed tumor initiation and development through the NF- B signaling pathway. These results indicate that miR-449a functions as a tumor suppressor in NSCLC by targeting the HMGB1-mediated NF- B signaling pathway in NSCLC.     

NF1与Twist在胃组织中的表达及其与临床病理相关性的研究

[目的]探讨神经纤维瘤基因(NF1)和转录因子Twist在不同胃组织中的表达状态及意义.[方法]应用免疫组织化学的方法检测NF1和Twist在87例胃癌组织、42例正常胃组织中的表达,比较两者的相关性.[结果] NF1在正常胃组织中阳性率10.3％,胃癌组织中为64.3％,差异有统计学意义(P＜0.05);Twist在正常胃组织中阳性率5.0％,胃癌组织中为68.6％,差异有统计学意义(P＜0.01).胃癌组织中NF1与Twist的表达具有负相关性(P＜0.01).NF1高表达组5年生存率明显高于低表达组(52.1％vs 17.6％);Twist高表达组5年生存率显著低于低表达组(17.9％vs 47.8％).[结论]Twist蛋白在胃癌组织中高表达,NF1蛋白低表达,它们的表达水平与胃癌患者的预后相关.