Circ_0001955 promotes non-small cell lung cancer cell proliferation and invasion by regulating MiR-29a-3p/NKIRAS2 axis to activate the nuclear factor-κB pathway

Non-small cell lung cancer (NSCLC) is one of the most common malignant tumors worldwide. Circular RNAs (circRNAs) have been widely reported to play a role in the pathogenesis of various tumors. Nevertheless, the function of circ_0001955 in NSCLC progression has not been explored yet. This study aims to explore the functions of circ_0001955 in NSCLC and investigate its regulatory molecular mechanism. First, we determined that circ_0001955 was upregulated in NSCLC cells. Subsequently, we demonstrated that knockdown of circ_0001955 restrained cell proliferation and invasion. In vivo experiments further proved the suppressive effect of circ_0001955 silence on tumor growth. Mechanism assays revealed that circ_0001955 enhanced nuclear factor-κB (NF-κB) inhibitor interacting Ras-like protein 2 (NKIRAS2) expression by sponging microRNA-29a-3p (miR-29a-3p). Upregulation of NKIRAS2 led to the deceased level of IκBβ but increased levels of nuclear p65, thus activating the NF-κB signaling pathway. In conclusion, Circ_0001955 activates the NF-κB pathway to promote NSCLC cell proliferation and invasion by regulating miR-29a-3p/NKIRAS2 axis.     

Downregulation of circ_0000673 Promotes Cell Proliferation and Migration in Endometriosis via the Mir-616-3p/PTEN Axis

Endometriosis is a common gynecological disease, affecting up to 10% of women of reproductive age and approximately 50% of women with infertility. Circular RNAs (circRNAs) have been shown to be involved in a number of diseases. Dysregulated expression of circRNAs in endometriosis has been reported, and circ_0000673 was significantly downregulated. However, the details of its role in the pathogenesis of endometriosis are still poorly understood. We investigated the location and effects of the downregulation of circ_0000673 in endometriosis. We demonstrated that knockdown of circ_0000673 significantly increased the proliferation and migration of eutopic and normal endometrial cells. Bioinformatics analysis predicted that circ_0000673 might act as a sponge for miR-616-3p. We found that the effect of circ_0000673 knockdown could be recovered by miR-616-3p inhibitor and enhanced by miR-616-3p mimics. qPCR and western blot assays showed that circ_0000673 knockdown could decrease the expression of PTEN and increase the expression of PI3K and p-AKT. PTEN was confirmed to be a target of miR-616-3p. These results demonstrated that the downregulation of circ_0000673 could promote the progression of endometriosis by inactivating PTEN via the deregulation of miR-616-3p.     

MiR-193a-3p and miR-193a-5p suppress the metastasis of human osteosarcoma cells by down-regulating Rab27B and SRR,respectively

MicroRNAs have been identified as key players in the development and progression of osteosarcoma, which is the most common primary malignancy of bone. Sequencing-based miR-omic and quantitative real-time PCR analyses suggested that the expression of miR-193a-3p and miR-193a-5p was decreased by DNA methylation at their promoter region in a highly metastatic osteosarcoma cell line (MG63.2) relative to their expression in the less metastatic MG63 cell line. Further wound-healing and invasion assays demonstrated that both miR-193a-3p and miR-193a-5p suppressed osteosarcoma cell migration and invasion. Moreover, introducing miR-193a-3p and miR-193a-5p mimics into MG63.2 cells or antagomiRs into MG63 cells confirmed their critical roles in osteosarcoma metastasis. Additionally, bioinformatics prediction along with biochemical assay results clearly suggested that the secretory small GTPase Rab27B and serine racemase (SRR) were direct targets of miR-193a-3p and miR-193a-5p, respectively. These two targets are indeed involved in the miR-193a-3p- and miR-193a-5p-induced suppression of osteosarcoma cell migration and invasion. MiR-193a-3p and miR-193a-5p play important roles in osteosarcoma metastasis through down-regulation of the Rab27B and SRR genes and therefore may serve as useful biomarkers for the diagnosis of osteosarcoma and as potential candidates for the treatment of metastatic osteosarcoma.     

Circ_0000218 plays a carcinogenic role in laryngeal cancer through regulating microRNA-139-3p/Smad3 axis

BackgroundLaryngeal squamous cell carcinoma (LSCC) accounts for about 85%–90% of all cases of laryngeal cancer. So far, the role and molecular mechanism of circular RNA 0,000,218 (circ_0000218)/microRNA (miR)-139−3p in laryngeal cancer are not clear. The present study aimed to investigate the role and regulatory mechanism of circ_0000218/miR-139−3p in laryngeal cancerin vitro and in vivo.Methodsquantitative real time polymerase chain reaction (qRT-PCR) was used to detect the expression of circ_0000218/miR-139−3p in LSCC cells. Dual luciferase reporter assay and RNA immunoprecipitation (RIP) assay were used to confirm binding sites between miR-139−3p and smad family member 3 (Smad3), and circ_0000218 and miR-139−3p. Cell Counting Kit-8 (CCK-8) and cell apoptosis analysis were used to detect cell viability and apoptosis. Xenograft experiment was performed to show in vivo effect of circ_0000218/miR-139−3p on the growth of LSCC.ResultsCirc_0000218 was highly expressed in LSCC cells. miR-139−3p, lower expressed in LSCC cells, was negatively regulated by circ_0000218 in LSCC cells. Besides, the findings suggested that circ_0000218 silencing inhibited the LSCC cell viability and promoted apoptosis by negatively regulating miR-139−3p expression. Furthermore, the data indicated that miR-139−3p inhibited the viability of LSCC cells and promoted apoptosis, and these effects were reversed by Smad3 over-expression. In addition, the in vivo effects of circ_0000218/miR-139−3p on LSCC were consistent with the in vitro study.Conclusionscirc_0000218 inhibition inhibited the growth of LSCC by targeting miR-139−3p/Smad3 axis. Our present study provided a new target for laryngeal cancer treatment.     

下调lncRNA DNM3OS通过靶向miR-193a-3p增强人直肠癌细胞系SW-480的放疗敏感性

目的 探讨lncRNA DNM3OS对直肠癌SW-480细胞的增殖、凋亡以及放射敏感性的影响及分子机制.方法 选取30例直肠癌患者癌组织及癌旁组织,RT-qPCR检测DNM3OS和miR-193a-3p的表达水平;将抑制DNM3OS的表达载体、过表达miR-193a-3p载体转染至SW-480细胞,将DNM3OS与抑制...     

miR-27a-3p靶向HOXA5基因通过Wnt/β-catenin信号通路调控糖尿病患者创面愈合

目的:研究微小RNA-27a-3p(miR-27a-3p)对糖尿病患者创面愈合的影响及作用机制.方法:采用qPCR和Western blot检测糖尿病患者创面愈合组织中miR-27a-3p、同源异型盒基因A5(HOXA5)mRNA和HOXA5蛋白的表达.使用高糖处理人微血管内皮细胞(HMECs),模拟糖尿病HMECs的...     

Circular RNA hsa_circ_0001598 promotes programmed death-ligand-1-mediated immune escape and trastuzumab resistance via sponging miR-1184 in breast cancer cells

Approximately 25% of breast cancer (BC) patients are HER2-positive. Trastuzumab is used as a targeted therapy drug to treat HER2-positive BC patients; however, the drug resistance remains a big challenge. Circular RNAs (circRNAs) are reported to be involved in drug resistance, but the role of circ_0001598 has never been studied in BC. First, we identified the expression of circ_0001598 by RT-qPCR in BC. The gain-of-function and loss-of-function studies were applied to study the functional roles of circ_0001598 and its target gene. We observed upregulation of circ_0001598 in BC tissues, especially in trastuzumab-resistant BC samples. We further identified that miR-1184 is a functional target of circ_0001598. Moreover, it was found that programmed death-ligand 1 (PD-L1) was a direct target of miR-1184. The oncogenic effects of circ_0001598 in promoting BC cell growth, trastuzumab-resistance, PD-L1 expression, and escaping of CD8 T cell killing were abolished after the restoration of miR-1184. In conclusion, we demonstrate that circ_0001598/miR-1184/PD-L1 signaling plays a crucial role in the regulation of BC progression and trastuzumab-resistance phonotypes, which suggests that circ_0001598 may be a molecular target to treat HER2-positive BC patients.

     

Circular RNA circEGFR regulates tumor progression via the miR-106a-5p/DDX5 axis in colorectal cancer

Recently, an increasing number of studies have reported that dysregulation of circular RNA (circRNA) expression plays critical roles in the progression of several cancers, including colorectal cancer (CRC). However, the detailed molecular mechanisms of circRNAs involvement in CRC remain largely unknown. Here, we confirmed that the level of circEGFR was significantly increased in CRC tissues compared to matched adjacent non-tumor tissues, and a high level of circEGFR was correlated with poor clinicopathological characteristics and poor prognosis in patients with CRC. Moreover, increased circEGFR expression promoted CRC cell proliferation, migration, and invasion in vitro. Mechanistically, circEGFR acted as a ceRNA for miR-106a-5p to relieve the repressive effect of miR-106a-5p on DDX5 mRNA. Moreover, circEGFR enhanced DDX5 expression, thereby upregulating p-AKT levels. Together, these findings showed that circEGFR promoted CRC cell proliferation, migration, and invasion through the miR-106a-5p/DDX5/AKT axis, and may serve as a promising diagnostic marker and therapeutic target for CRC patients.     

Cyclic RNA Circ_0000735 sponges miR-502-5p to promote bladder cancer cell proliferation and invasion and inhibit apoptosis

Objective: The objective of this study was to investigate the effect on the proliferation, invasion, and apoptosis of bladder cancer cells through miR-502-5p of the Circ_0000735 circular RNA. Methods: Circ_0000735 and miR-502-5p expression of bladder cancer patients in malignant and paracancerous tissues was identified using qRT-PCR. Nucleoplasm isolation assay and RNase R enzymatic assay were used to classify Circ_0000735 subcellular origin and stability. Dual luciferase reporter assay and RIP assay were used to confirm Circ_0000735 and miR-502-5p targeting relationships. Cell proliferation, apoptosis, and invasion capacity were identified using CCK8, flow cytometry, and transwell assays. To confirm the effect of Circ_0000735 on tumorigenesis in nude mice, in vivo experiments were conducted. Results: Circ_0000735 expression was increased in bladder cancer tissues and cells compared with paraneoplastic tissues and normal cells, and miR-502-5p expression was reduced (both P<0.05). In the cytoplasm, Circ_0000735 was largely clustered and could not be digested by the RNase R enzyme, and ceRNA may play a role in bladder cancer cells. Circ_0000735 silencing prevented cell proliferation and invasion and facilitated apoptosis (all P<0.05). The incorporation of miR-502-5p inhibitor rescued the effect on bladder cancer cells of Circ_0000735 silencing. In vitro experiments showed that inhibition of Circ_0000735 expression was beneficial in suppressing tumorigenic ability in nude mice. Conclusion: Circ_0000735 can adsorb miR-502-5p to promote bladder cancer cell proliferation and invasion and inhibit apoptosis. Circ_0000735 may be an effective molecular target for bladder cancer therapy.     

DNM3OS Facilitates Ovarian Cancer Progression by Regulating miR-193a-3p/MAP3K3 Axis

PurposeLong non-coding RNAs (lncRNAs) are essential regulators in the development of ovarian cancer (OC). Nonetheless, the function of lncRNA DNM3 opposite strand/antisense RNA (DNM3OS) in OC remains unclear. This work aimed to investigate the biological roles and underlying mechanisms of DNM3OS in OC.Materials and MethodsQuantitative real-time polymerase chain reaction was conducted to examine DNM3OS, microRNA (miR)-193a-3p, and mitogen-activated protein kinase 3 (MAP3K3) mRNA expression in OC tissues and cell lines. Kaplan-Meier survival analysis was employed to analyze the relationship between DNM3OS expression and the prognosis of OC patients. Cell counting kit-8, 5-ethynyl-2′-deoxyuridine, and transwell experiments were conducted to monitor cell proliferation, migration, and invasion, respectively. Western blot was applied to examine epithelial-mesenchymal transition associated protein (E-cadherin and N-cadherin) expression. Luciferase reporter gene and RNA immunoprecipitation experiments were performed to confirm the relationships among DNM3OS, miR-193a-3p, and MAP3K3. Pearson''s correlation analysis was adopted to analyze the correlations among DNM3OS, miR-193a-3p, and MAP3K3 mRNA.ResultsDNM3OS expression was remarkably increased in OC tissues and cell lines, which was associated with the unfavorable prognosis of the patients. DNM3OS overexpression enhanced OC cell proliferation, migration, and invasion; suppressed E-cadherin protein expression; and facilitated N-cadherin protein expression, while the transfection of miR-193a-3p mimics had the opposite effects. DNM3OS directly interacted with miR-193a-3p, and miR-193a-3p targeted MAP3K3 by directly binding to 3′UTR. DNM3OS could up-regulate the expression of MAP3K3 via repressing miR-193a-3p expression.ConclusionDNM3OS, as an oncogenic lncRNA, increases the malignancy of OC cells via regulation of an miR-193a-3p/MAP3K3 axis.     

miR-338-3p suppresses invasion of liver cancer cell by targeting smoothened

MicroRNAs are involved in human carcinogenesis and cancer progression. Our previous study has shown that loss of miR-338-3p expression is associated with clinical aggressiveness of hepatocellular carcinoma (HCC). However, the exact roles and mechanisms of miR-338-3p remain unknown in HCC. To determine whether and how miR-338-3p influences liver cancer cell invasion, we studied miR-338-3p in the liver cancer cell lines, and we found that miR-338-3p is down-regulated in treated cells. Forced expression of miR-338-3p in SK-HEP-1 cells suppressed cell migration and invasion, whereas inhibition of miR-338-3p in SMMC-7721 cells induced cell migration and invasion. Furthermore, smoothened (SMO) was identified as a direct target of miR-338-3p. Forced expression of miR-338-3p down-regulated SMO and matrix metalloproteinase (MMP)-9 expression, but inhibition of miR-338-3p up-regulated SMO and MMP9 expression. However, small interfering RNA targeted SMO reversed the effects induced by blockade of miR-338-3p. SMO and MMP9 were overexpressed and associated with invasion and metastasis in HCC tissues. These data indicate that miR-338-3p suppresses cell invasion by targeting the smoothened gene in liver cancer in vitro and miR-338-3p might be a novel potential strategy for liver cancer treatment.     

RETRACTED: CircFBXW7 alleviates glioma progression through regulating miR-23a-3p/PTEN axis

Increasing evidence has confirmed that circular RNAs (circRNAs) are involved in regulating the development and progression of various tumors. The aim of this study was to examine the effect of circFBXW7 on the progression of glioma and to determine its underlying mechanism. qRT-PCR was performed to measure the expression of circFBXW7, miR-23a-3p, and PTEN in tissues and cell lines of glioma. The proliferation ability of glioma cells was examined using the CCK-8 assay. Glioma cell migration and invasion capacity were detected using Transwell assays. The dual-luciferase reporter gene assay was employed to examine the correlation between miR-23a-3p and circFBXW7 or PTEN. The expression levels of the related genes were determined using western blotting analysis. A glioma xenograft tumor model was employed to evaluate the functional roles of circFBXW7 in vivo. CircFBXW7 was found to be aberrantly downregulated in glioma tumor tissues and cell lines. Overexpression of circFBXW7 was found to significantly inhibit the proliferation, migration and invasion ability of the glioma cells. Moreover, bioinformatic analysis and dual-luciferase reporter assays confirmed that circFBXW7 can directly target miR-23a-3p, which then blocks the binding of miR-23a-3p to the 3′ un-translated region (UTR) of PTEN. Mechanically, circFBXW7 suppresses cell proliferation and metastasis in glioma by sponging miR-23a-3p, resulting in elevated PTEN expression. In addition, in vivo experiments also confirmed that circFBXW7 overexpression effectively halts tumor growth and metastasis. Consistent with the in vitro observations, circFBXW7 overexpression significantly decreased miR-23a-3p, Ki-67, and N-cadherin, as well as increased PTEN and E-cadherin levels. Our results revealed that circFBXW7 exhibits antiproliferative and antimetastasis activities via sponging miR-23a-3p to elevate PTEN expression in glioma, which may offer a novel target for clinical therapy and diagnosis of glioma.     

Profilin 1, negatively regulated by microRNA-19a-3p,serves as a tumor suppressor in human hepatocellular carcinoma

Profilin 1 (PFN1) is a critical actin-regulatory protein; however, its functional role in hepatocellular carcinoma (HCC) progression remains to be further elucidated. In the present study, we observed that the expression levels of PFN1 were significantly decreased in HCC tissues and cell lines. Low PFN1 expression was significantly correlated with aggressive clinicopathological characteristics and poor prognosis of HCC patients. Further in vitro experiments demonstrated that overexpression of PFN1 remarkably inhibited the proliferation, migration, invasion and EMT of HCC cells. Moreover, we also found that PFN1 was a direct target gene of miR-19a-3p, and in HCC tissues, and there was a significantly inverse correlation between PFN1 mRNA and miR-19a-3p expression. Collectively, our results showed that PFN1 functions as a tumor suppressor in HCC, and might serve as a diagnostic and therapeutic target for HCC patients.     

MiR-199-3p replacement affects E-cadherin expression through Notch1 targeting in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) represents the second cause of cancer-related mortality worldwide and is associated with poor prognosis, due to a high recurrence rate after curative treatments and a drug resistance phenotype. In this scenario, the identification of innovative and effective therapeutic strategies is an unmet clinical need. The safety and efficacy of microRNA (miRNA) mediated approaches in preclinical models and clinical trials have been widely described in cancer. MicroRNA-199a downregulation is a common feature of HCC where its reduced expression contributes to mTOR and c-Met pathways activation. Notch1 activation is also a common event in HCC, influencing epithelial-to-mesenchymal transition, tumor invasion and recurrence at least in part through E-cadherin regulation. Here we identified a negative correlation between miR-199a-3p and Notch1 or E-cadherin protein levels in HCC patients and demonstrated that miR-199a-3p regulates E-cadherin expression through Notch1 direct targeting in in vitro models. Moreover, we showed that a strong correlation exists between miR-199a-5p and miR-199a-3p in HCC specimens and that miR-199a-5p contributes to E-cadherin regulation as well, underlying the complex network of interaction carried out by miR-199a and its influence on tumor aggressiveness. In conclusion, our findings suggest the restoration of miR-199a-3p physiologic levels as a possible therapeutic strategy for the treatment of HCC.     

Dysregulation of microRNA-106a-5p-RUNX1 axis associates with clinical progression and prognosis of osteosarcoma patients

MicroRNA-106a-5p (miR-106a-5p) functions as a tumor suppressor in osteosarcoma cells. Here, we aimed to identify novel target genes of miR-106a-5p in osteosarcoma, as well as to investigate their prognostic value and the biological functions. At first, the mammalian runt-related factor 1 (RUNX1) was identified as one of the target genes of miR-106a-5p in osteosarcoma cells by luciferase reporter gene assay, real-time quantitative RT-PCR and Western blot analysis. Then, the expression levels of miR-106a-5p and RUNX1 in osteosarcoma tissues were detected, and their associations with clinicopathological features and patients' prognosis were statistically analyzed. Compared with adjacent non-cancerous tissues, miR-106a-5p and RUNX1 mRNA/protein expression in osteosarcoma tissues were significantly decreased and increased, respectively (all P < 0.01). Low miR-106a-5p, high RUNX1 and miR-106a-5p-low/RUNX1-high expression in osteosarcoma tissues were all significantly associated with advanced Enneking stage, positive metastasis and shorter overall survival (all P < 0.05). Moreover, miR-106a-5p and RUNX1 expression, alone or in combination, were identified as independent prognostic factors for osteosarcoma patients' overall survival. Functionally, the enforced expression of miR-106a-5p significantly suppressed proliferation and invasion of osteosarcoma cells, while the overexpression of RUNX1 effectively reversed its suppressive roles. In conclusion, our findings show the dysregulation of miR-106a-5p-RUNX1 axis in human osteosarcoma tissues and suggest its crucial roles in cancer progression and patients' prognosis. More interestingly, miR-106a-5p may function as a tumor suppressor in osteosarcoma cells via regulating its target gene RUNX1.     

喉癌组织中的microRNA差异表达谱及miR-125a-5p抑制喉癌细胞增殖的初步研究

 目的：筛选并分析喉癌组织与周围正常喉黏膜的微小RNA(microRNAs,miRNAs) 之间的表达谱差异,为进一步研究miRNA与喉癌发生、发展的关系提供线索。方法：收集喉癌组织和癌旁正常喉黏膜标本共42对,随机选取10对标本进行miRNA微阵列基因芯片分析, 另选取32对标本进行实时荧光定量PCR （qRT-PCR）验证,获得喉癌组织中的miRNA差异表达谱。应用MTT法和克隆形成实验检测miR-125a-5p对喉癌Hep2细胞增殖的影响。结果：喉癌组织中的let-7f-5p、miR-10a-5p、miR-125a-5p、miR-144-3p、miR-195-5p、miR-203等6个miRNA在基因芯片以及qRT-PCR中表达均显著下调。与对照组相比,转染miR-125a-mimics组的喉癌Hep2细胞增殖能力受到抑制,而转染miR-125a-inhibitor组Hep2细胞增殖能力增强。结论：基因芯片与qRT-PCR结果一致;喉癌与正常喉黏膜之间存在明显的miRNA差异表达,这些miRNA的差异性表达可能与喉癌的发病、侵袭等相关。miR-125a可以抑制喉癌Hep2细胞的增殖,可能作为喉癌生物治疗的新靶点。     

miR-188-3p靶向YAP1抑制肝细胞癌的侵袭和迁移

目的 探究miR-188-3p在肝细胞癌(hepatocellular carcinoma,HCC)中的表达及其抑制HCC细胞侵袭和迁移的分子机制.方法 利用RT-qPCR检测miR-188-3p在人HCC组织和细胞系中的表达情况,进一步分析miR-188-3p表达在HCC中的临床价值;利用Western印迹检测转染m...