CircSNHG5 Sponges Mir-495-3p and Modulates CITED2 to Protect Cartilage Endplate From Degradation

BackgroundIntervertebral disc degeneration (IDD) is a highly prevalent degenerating disease that produces tremendous amount of low back and neck pain. The cartilage endplate (CEP) is vitally important to intervertebral discs in both physiological and pathological conditions. In addition, circular RNAs (circRNAs) have been shown to be involved in the regulation of various diseases, including IDD. However, the particular role of circRNAs in cervical vertebral CEP degeneration remains unclear. Here, we examined the unique role of circRNAs in CEP of patients with cervical fracture and degenerative cervical myelopathy (DCM).MethodsHuman competitive endogenous RNA (ceRNA) microarray was performed by previous research. Western blot (WB), immunofluorescence (IF), quantitative RT-PCR (qRT-PCR), luciferase assay, and fluorescence in situ hybridization (FISH) were employed to analyze the function of circSNHG5 and its downstream effectors, miR-495-3p, and CITED2.ResultsWe demonstrated that circSNHG5 expression was substantially low in degenerative CEP tissues. Knockdown of circSNHG5 in chondrocytes resulted in a loss of cell proliferation and followed by degradation of extracellular matrix (ECM). In addition, circSNHG5 was shown to sponge miR-495-3p and modulate the expression of the downstream gene CITED2. This mechanism of action was further validated via overexpression and knockdown of CITED2.ConclusionOur findings identified a novel circSNHG5-miR-495-3p axis responsible for IDD progression. Future investigations into IDD therapy may benefit from targeting this axis.

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Roles of ncRNAs as ceRNAs in Gastric Cancer

Genes ◽

10.3390/genes12071036 ◽

2021 ◽

Vol 12 (7) ◽

pp. 1036

Author(s):

Junhong Ye ◽

Jifu Li ◽

Ping Zhao

Keyword(s):

Gastric Cancer ◽

Research Strategy ◽

Circular Rnas ◽

Messenger Rnas ◽

Cerna Network ◽

Degree Of Malignancy ◽

High Degree ◽

Competitive Endogenous Rna ◽

Made In

Although ignored in the past, with the recent deepening of research, significant progress has been made in the field of non-coding RNAs (ncRNAs). Accumulating evidence has revealed that microRNA (miRNA) response elements regulate RNA. Long ncRNAs, circular RNAs, pseudogenes, miRNAs, and messenger RNAs (mRNAs) form a competitive endogenous RNA (ceRNA) network that plays an essential role in cancer and cardiovascular, neurodegenerative, and autoimmune diseases. Gastric cancer (GC) is one of the most common cancers, with a high degree of malignancy. Considerable progress has been made in understanding the molecular mechanism and treatment of GC, but GC’s mortality rate is still high. Studies have shown a complex ceRNA crosstalk mechanism in GC. lncRNAs, circRNAs, and pseudogenes can interact with miRNAs to affect mRNA transcription. The study of the involvement of ceRNA in GC could improve our understanding of GC and lead to the identification of potential effective therapeutic targets. The research strategy for ceRNA is mainly to screen the different miRNAs, lncRNAs, circRNAs, pseudogenes, and mRNAs in each sample through microarray or sequencing technology, predict the ceRNA regulatory network, and, finally, conduct functional research on ceRNA. In this review, we briefly discuss the proposal and development of the ceRNA hypothesis and the biological function and principle of ceRNAs in GC, and briefly introduce the role of ncRNAs in the GC’s ceRNA network.

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Hsa_circ_0051079 functions as an oncogene by regulating miR-26a-5p/TGF-β1 in osteosarcoma

Cell & Bioscience ◽

10.1186/s13578-019-0355-2 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 7

Author(s):

Zuojun Zhang ◽

Ming Zhao ◽

Guojie Wang

Keyword(s):

Luciferase Activity ◽

Bioinformatic Analysis ◽

Luciferase Assay ◽

Circular Rnas ◽

Potential Biomarker ◽

Luciferase Activity Assay ◽

Proliferation And Metastasis ◽

Normal Controls ◽

Tgf Β1

Abstract Background Osteosarcoma is a most common bone malignant tumor which threatens children and adolescents. Circular RNAs (circRNAs) fundamentally play essential roles in the progress and development of human cancers by sponging with microRNAs (miRNAs). However, the role of circRNAs in osteosarcoma is not clear. The aim of the study was to investigate the roles and molecular mechanism of circRNAs in osteosarcoma. Results The data from qRT-PCR showed that circ_0051079 expression was higher in osteosarcoma cells and tissues compared to their normal controls. Meanwhile, bioinformatic analysis indicated that circ_0051079 was a sponge of miR-26a-5p, which was verified by luciferase activity assay. Subsequently, TGF-β1 was verified as a putative target mRNA of miR-26a-5p by luciferase assay. Cellular function assays were conducted and the findings revealed that circ_0051079/miR-26a-5p/TGF-β1 regulated osteosarcoma proliferation and metastasis. Conclusion The study demonstrated that circ_0051079 could act as an oncogene via regulating miR-26a-5p/TGF-β1 and a potential biomarker for osteosarcoma diagnose.

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Circular RNA hsa_circ_0001829 promotes gastric cancer progression through miR-155-5p/SMAD2 axis

Journal of Experimental & Clinical Cancer Research ◽

10.1186/s13046-020-01790-w ◽

2020 ◽

Vol 39 (1) ◽

Author(s):

Qiuling Niu ◽

Zhijie Dong ◽

Min Liang ◽

Yuanwei Luo ◽

Hai Lin ◽

...

Keyword(s):

Gastric Cancer ◽

Cell Lines ◽

Cancer Progression ◽

Cell Cycle Progression ◽

Luciferase Assay ◽

Migration And Invasion ◽

Circular Rnas ◽

Gain Of Function

Abstract Background Accumulating evidences have shown that circular RNAs (circRNAs) play important roles in regulating the pathogenesis of cancer. However, the role of circRNAs in gastric cancer (GC) remains largely unclear. Methods In this study, we identified a novel upregulated circRNA, hsa_circ_0001829, in chemically induced malignant transformed human gastric epithelial cells using RNA-seq. Subsequent qRT-PCR and ISH assays were performed to detect the expression level of hsa_circ_0001829 in GC cell lines and tissues. Functional roles of hsa_circ_0001829 in GC were then explored by loss- and gain-of- function assays. Bioinformatic prediction and luciferase assay were used to investigate potential mechanisms of hsa_circ_0001829. Finally, the mice xenograft and metastasis models were constructed to assess the function of hsa_circ_0001829 in vivo. Results We found that hsa_circ_0001829 was significantly upregulated in GC tissues and cell lines. Loss- and gain-of- function assays showed that hsa_circ_0001829 promotes GC cells proliferation, migration and invasion, and the affected cell cycle progression and apoptosis rates may account for the effect of hsa_circ_0001829 on GC proliferation. In addition, bioinformatic prediction and luciferase assay showed that hsa_circ_0001829 acts as a molecular sponge for miR-155-5p and that SMAD2 was a target gene of miR-155-5p; moreover, hsa_circ_0001829 sponges miR-155-5p to regulate SMAD2 expression and hsa_circ_0001829 promotes GC progression through the miR-155-5p–SMAD2 pathway. Finally, suppression of hsa_circ_0001829 expression inhibited tumor growth and aggressiveness in vivo. Conclusion Taken together, our findings firstly demonstrated a novel oncogenic role of hsa_circ_0001829 in GC progression through miR-155-5p–SMAD2 axis, and our study may offer novel biomarkers and therapeutic targets for GC.

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Inhibition of intervertebral disc disease progression via the circPKNOX1–miR-370-3p–KIAA0355 axis

Cell Death Discovery ◽

10.1038/s41420-021-00420-4 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Yizhen Huang ◽

Jun Gao ◽

Jianle Wang ◽

Huali Ye ◽

Teng Yao ◽

...

Keyword(s):

Intervertebral Disc ◽

Target Genes ◽

Circular Rnas ◽

Intervertebral Disc Disease ◽

Matrix Synthesis ◽

Immunofluorescence Analysis ◽

Disc Disease ◽

Compensatory Response

AbstractThe molecular mechanism underlying the development of intervertebral disc disease (IVDD) is not completely understood. Circular RNAs (circRNAs) play a significant role in the occurrence and development of various diseases, and studies have shown that circPKNOX1 is involved in the compensatory response of extracellular matrix synthesis and secretion of the nucleus pulposus (NP) cells. However, the mechanism through which circRNAs regulate IVDD progression remains unclear; therefore, in this study, we explored the significance of circPKNOX1 in IVDD. The expression of circRNAs in NP cells of normal and degenerative patients was detected using microarray analysis, and the role of circPKNOX1 in IVDD was confirmed using RT-qPCR. The interaction networks of circRNAs, miRNAs, and miRNA target genes were detected using bioinformatics analysis, RNA fluorescence in situ hybridization, and immunofluorescence analysis. We found that the expression of circPKNOX1 decreased in IVDD cells. The expression of circPKNOX1 in NP cells, observed using RT-qPCR and western blotting, was consistent with that observed using array screening. Overexpression of circPKNOX1 increased the expression of collagen II, aggrecan, and SOX9 and decreased that of ADAMTS4, ADAMTS-5, MMP3, and MMP13. We further demonstrated that circPKNOX1 played the role of a sponge by competitively binding miR-370-3p to reverse the inhibition of KIAA0355 expression. Our findings indicated that circPKNOX1 affected the progression of IVDD by regulating the expression of KIAA0355 via miR-370-3p. Therefore, circPKNOX1-based therapy may serve as an effective IVDD treatment strategy.

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lncRNA IGHCγ1 Acts as a ceRNA to Regulate Macrophage Inflammation via the miR-6891-3p/TLR4 Axis in Osteoarthritis

Mediators of Inflammation ◽

10.1155/2020/9743037 ◽

2020 ◽

Vol 2020 ◽

pp. 1-11

Author(s):

Pengjun Zhang ◽

Jianmei Sun ◽

Caihong Liang ◽

Bingjie Gu ◽

Yang Xu ◽

...

Keyword(s):

Inflammatory Response ◽

Noncoding Rna ◽

Luciferase Reporter ◽

Mirna Sponge ◽

And Migration ◽

And Function ◽

Competitive Endogenous Rna ◽

Proliferation And Migration

Accumulating data have implicated that long noncoding RNA (lncRNA) plays an important role in osteoarthritis (OA), which may function as a competitive endogenous RNA (ceRNA) of microRNAs (miRNAs). lncRNA IGHCγ1 has been demonstrated to regulate inflammation and autoimmunity. Nonetheless, the altering effect of IGHCγ1 in OA remains unclear. This study is aimed at investigating the mechanism and function of lncRNA IGHCγ1 in OA. CCK-8, EdU, and transwell assays were used to estimate macrophage proliferation and migration. Fluorescence in situ hybridization (FISH) was performed to estimate the local expression of lncRNA IGHCγ1 in macrophages. Luciferase reporter assay was adopted to validate the ceRNA role of IGHCγ1 as miRNA sponge. lncRNA IGHCγ1 was primarily localized in macrophage cytoplasm and upregulated in OA. miR-6891-3p inhibited macrophage proliferation, migration, and inflammatory response by targeting TLR4, while lncRNA IGHCγ1 promoted TLR4 expression by functioning as a ceRNA for miR-6891-3p through the NF-κB signal in macrophages. This study strongly supports that lncRNA IGHCγ1 regulates inflammatory response via regulating the miR-6891-3p/TLR4/NF-κB axis in macrophages.

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CircTUBGCP3 facilitates the tumorigenesis of lung adenocarcinoma by sponging miR-885-3p

10.21203/rs.3.rs-502804/v1 ◽

2021 ◽

Author(s):

Yang Yang ◽

Xin Fan ◽

Yunfei Nie ◽

Donglei Liu ◽

Dengyan Zhu ◽

...

Keyword(s):

Lung Adenocarcinoma ◽

Clinicopathological Characteristics ◽

Circular Rnas ◽

Luciferase Gene ◽

Poor Survival ◽

Rna Immunoprecipitation ◽

Underlying Mechanisms

Abstract Background: Circular RNAs (circRNAs) act pivotal roles in the progression of multiple malignancies. However, the underlying mechanisms by which hsa_circ_0007031 (circTUBGCP3) contributes to lung adenocarcinoma (LAC) remain largely unknown.Methods: The association of circTUBGCP3 expression with clinicopathological characteristics and prognosis in patients with LAC was determined by RT-qPCR and fluorescence in situ hybridization. The in vitro functional experiments as well as a subcutaneous tumorigenesis model were executed to estimate the role of circTUBGCP3 in LAC cells. The interaction between circTUBGCP3 and miR-885-3p was confirmed by RNA immunoprecipitation, luciferase gene report and RT-qPCR assays. The effects of circTUBGCP3 on miR-885-3p-mediated Wnt10b/β-catenin signaling were evaluated by Western blot. Results: The upregulation of circTUBGCP3 or downregulation of miR-885-3p was associated with the pathological stage and poor survival in patients with LAC. Restored expression of circTUBGCP3 facilitated the growth and invasion of LAC cells, but knockdown of circTUBGCP3 harbored the opposite effects. In mechanism, circTUBGCP3 could act as a sponge of miR-885-3p, which suppressed the cell proliferation and colony formation and attenuated the tumor-promoting effects of circTUBGCP3. Wnt10b as a target of miR-885-3p could be upregulated be circTUBGCP3 and indicate poor survival in patient with LAC. Conclusions: Our findings demonstrated that circTUBGCP3 promoted LAC progression by sponging miR-885-3p, and might represent a prognostic factor for LAC.

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Circular RNA hsa_circ_0001829 Promotes Gastric Cancer Progression through miR-155-5p/SMAD2 Axis

10.21203/rs.3.rs-57293/v1 ◽

2020 ◽

Author(s):

Qiuling Niu ◽

Zhijie Dong ◽

Min Liang ◽

Yuanwei Luo ◽

Hai Lin ◽

...

Keyword(s):

Gastric Cancer ◽

Cell Lines ◽

Cancer Progression ◽

Cell Cycle Progression ◽

Luciferase Assay ◽

Migration And Invasion ◽

Circular Rnas ◽

Gain Of Function

Abstract Background: Accumulating evidences have shown that circular RNAs (circRNAs) play important roles in regulating the pathogenesis of cancer. However, the role of circRNAs in gastric cancer (GC) remains largely unclear. Methods: In this study, we identified a novel upregulated circRNA, hsa_circ_0001829, in chemically induced malignant transformed human gastric epithelial cells using RNA-seq. Subsequent qRT-PCR was performed to detect the expression level of hsa_circ_0001829 in GC cell lines and tissues. Functional roles of hsa_circ_0001829 in GC were then explored by loss- and gain-of- function assays. Bioinformatic prediction and luciferase assay were used to investigate potential mechanisms of hsa_circ_0001829. Finally, the mice xenograft models were constructed to assess the function of hsa_circ_0001829 in vivo.Results: We found that hsa_circ_0001829 was significantly upregulated in GC tissues and cell lines. Loss- and gain-of- function assays showed that hsa_circ_0001829 promotes GC cells proliferation, migration and invasion, and the affected cell cycle progression and apoptosis rates may account for the effect of hsa_circ_0001829 on GC proliferation. In addition, bioinformatic prediction and luciferase assay showed that hsa_circ_0001829 acts as a molecular sponge for miR-155-5p and that SMAD2 was a target gene of miR-155-5p; moreover, hsa_circ_0001829 sponges miR-155-5p to regulate SMAD2 expression and hsa_circ_0001829 promotes GC progression through the miR-155-5p–SMAD2 pathway. Finally, suppression of hsa_circ_0001829 expression inhibited subcutaneous tumor growth in vivo. Conclusion: Taken together, our findings firstly demonstrated a novel oncogenic role of hsa_circ_0001829 in GC progression, which is through miR-155-5p–SMAD2 axis, and our study may offer novel biomarkers and therapeutic targets for GC.

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CircTUBGCP3 facilitates the tumorigenesis of lung adenocarcinoma by sponging miR-885-3p

Cancer Cell International ◽

10.1186/s12935-021-02356-2 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Yang Yang ◽

Xin Fan ◽

Yunfei Nie ◽

Donglei Liu ◽

Dengyan Zhu ◽

...

Keyword(s):

Lung Adenocarcinoma ◽

Clinicopathological Characteristics ◽

Circular Rnas ◽

Luciferase Gene ◽

Poor Survival ◽

Rna Immunoprecipitation ◽

Underlying Mechanisms

Abstract Background Circular RNAs (circRNAs) act pivotal roles in the progression of multiple malignancies. However, the underlying mechanisms by which hsa_circ_0007031 (circTUBGCP3) contributes to lung adenocarcinoma (LAC) remain largely unknown. Methods The association of circTUBGCP3 expression with clinicopathological characteristics and prognosis in patients with LAC was determined by RT-qPCR and fluorescence in situ hybridization. The in vitro functional experiments as well as a subcutaneous tumorigenesis model were executed to estimate the role of circTUBGCP3 in LAC cells. The interaction between circTUBGCP3 and miR-885-3p was confirmed by RNA immunoprecipitation, luciferase gene report and RT-qPCR assays. The effects of circTUBGCP3 on miR-885-3p-mediated Wnt10b/β-catenin signaling were evaluated by Western blot. Results The upregulation of circTUBGCP3 or downregulation of miR-885-3p was associated with the pathological stage and poor survival in patients with LAC. Restored expression of circTUBGCP3 facilitated the growth and invasion of LAC cells, but knockdown of circTUBGCP3 harbored the opposite effects. In mechanism, circTUBGCP3 could act as a sponge of miR-885-3p, which suppressed the cell proliferation and colony formation and attenuated the tumor-promoting effects of circTUBGCP3. Wnt10b as a target of miR-885-3p could be upregulated be circTUBGCP3 and indicate poor survival in patient with LAC. Conclusions Our findings demonstrated that circTUBGCP3 promoted LAC progression by sponging miR-885-3p, and might represent a prognostic factor for LAC.

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circFAM160A2 Promotes Mitochondrial Stabilization and Apoptosis Reduction in Osteoarthritis Chondrocytes by Targeting miR-505-3p and SIRT3

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/5712280 ◽

2021 ◽

Vol 2021 ◽

pp. 1-13

Author(s):

Jiapeng Bao ◽

Changjian Lin ◽

Xing Zhou ◽

Diana Ma ◽

Lujie Ge ◽

...

Keyword(s):

Crucial Role ◽

Pathological Process ◽

Luciferase Assay ◽

Chondrocyte Apoptosis ◽

Circular Rnas ◽

Apoptosis Assay ◽

Safranin O

Competitive endogenous RNAs (ceRNAs), as a newly identified regulating mechanism, have been demonstrated to play a crucial role in various human diseases. An increasing number of recent studies have revealed that circular RNAs (circRNAs) can function as ceRNAs. However, little is known about the role of circFAM160A2 in the pathological process of osteoarthritis (OA). This study is the first to examine the crucial role of the circFAM160A2-miR-505-3p-SIRT3 axis in osteoarthritis progression. miR-505-3p was selected from the interaction of a microRNA (miRNA) microarray comparing chondrocytes in OA and normal conditions and prediction results from TargetScan. RT-qPCR was performed to assess the expression of circFAM160A2, miR-505-3p, and SIRT3. A dual luciferase assay was used to validate the binding of circFAM160A2, miR-505-3p, and SIRT3. We used lentivirus and adeno-associated virus to establish in vitro and in vivo overexpression models. Western blotting, apoptosis assay, ROS detection assay, Safranin O staining, and CCK-8 assay were employed to assess the role of circFAM160A2, miR-505-3p, and SIRT3. We found that miR-505-3p was upregulated and circFAM160A2 was downregulated in OA. While overexpression of circFAM160A2 decreased the production of extracellular matrix (ECM) degrading enzymes and ameliorated chondrocyte apoptosis and mitochondrial dysfunction, inhibition of miR-505-3p could reverse the protective effect of circFAM160A2 on the OA phenotype both in vitro and in vivo. In conclusion, circFAM160A2 can promote mitochondrial stabilization and apoptosis reduction in OA chondrocytes by targeting miR-505-3p and SIRT3, which might be a potential therapeutic target for OA therapy.

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A computer-control program for TEM in situ fatigue experiments

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100155074 ◽

1989 ◽

Vol 47 ◽

pp. 620-621

Author(s):

Kenneth S. Vecchio ◽

John A. Hunt

Keyword(s):

Slow Crack Growth ◽

Computer Control ◽

Control Program ◽

Video Tape ◽

Computer Control System ◽

Transmission Electron ◽

In Situ Experiments ◽

Tremendous Amount ◽

And Control

In-situ experiments conducted within a transmission electron microscope provide the operator a unique opportunity to directly observe microstructural phenomena, such as phase transformations and dislocation-precipitate interactions, “as they happen”. However, in-situ experiments usually require a tremendous amount of experimental preparation beforehand, as well as, during the actual experiment. In most cases the researcher must operate and control several pieces of equipment simultaneously. For example, in in-situ deformation experiments, the researcher may have to not only operate the TEM, but also control the straining holder and possibly some recording system such as a video tape machine. When it comes to in-situ fatigue deformation, the experiments became even more complicated with having to control numerous loading cycles while following the slow crack growth. In this paper we will describe a new method for conducting in-situ fatigue experiments using a camputer-controlled tensile straining holder.The tensile straining holder used with computer-control system was manufactured by Philips for the Philips 300 series microscopes. It was necessary to modify the specimen stage area of this holder to work in the Philips 400 series microscopes because the distance between the optic axis and holder airlock is different than in the Philips 300 series microscopes. However, the program and interfacing can easily be modified to work with any goniometer type straining holder which uses a penrmanent magnet motor.

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