scholarly journals The circular RNA circSlc7a11 promotes bone cancer pain pathogenesis in rats by modulating LLC-WRC 256 cell proliferation and apoptosis

2021 ◽  
Author(s):  
Han-Wen Chen ◽  
Xiao-Xia Zhang ◽  
Zhu-Ding Peng ◽  
Zu-Min Xing ◽  
Yi-Wen Zhang ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Cancer Pain ◽  
Expression Profiles ◽  
Bone Cancer ◽  
Circular Rna ◽  
Differentially Expressed ◽  
Bone Cancer Pain ◽  
Circular Rnas ◽  
Altered Expression ◽  
Proliferation And Apoptosis

AbstractTreatment of bone cancer pain (BCP) caused by bone metastasis in advanced cancers remains a challenge in clinical oncology, and the underlying mechanisms of BCP are poorly understood. This study aimed to investigate the pathogenic roles of circular RNAs (circRNAs) in regulating cancer cell proliferation and BCP development. Eight differentially expressed circRNAs in the rat spinal cord were validated by agarose gel electrophoresis and Sanger sequencing. Expression of circRNAs and mRNAs was detected by quantitative RT-PCR. MTS assay and flow cytometry were performed to analyze cell proliferation and apoptosis, respectively. Differentially expressed mRNA profiles were characterized by deep RNA sequencing, hierarchical clustering, and functional categorization. The interactions among circRNAs, microRNAs (miRNAs), and mRNAs were predicted using TargetScan. Additionally, western blot was performed to determine the protein levels of Pax8, Isg15, and Cxcl10. Multiple circRNAs were differentially expressed in the spinal cords of BCP model rats; of these, circSlc7a11 showed the greatest increase in expression. The overexpression of circSlc7a11 significantly promoted cell proliferation and repressed apoptosis of LLC-WRC 256 and UMR-106 cells, whereas circSlc7a11 silencing produced the opposite effects. Altered expression of circSlc7a11 also induced substantial changes in the mRNA expression profiles of LLC-WRC 256 cells; these changes were linked to multiple apoptotic processes and signaling pathways, such as the chemokine signaling pathway, and formed a complex circRNA/miRNA/mRNA network. Additionally, Pax8, Isg15, and Cxc110 protein level in LLC-WRC 256 cells was consistent with the mRNA results. The circRNA circSlc7a11 regulates rat BCP development by modulating LLC-WRC 256 cell proliferation and apoptosis through multiple-signaling mechanisms.

scholarly journals Transcriptomic Analysis of Long Noncoding RNA and mRNA Expression Profiles in the Amygdala of Rats with Bone Cancer Pain-Depression Comorbidity

Life ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 834
Author(s):  
Shuyan Wu ◽  
Xiaohui Chen ◽  
Fengyi Huang ◽  
Mingxue Lin ◽  
Pinzhong Chen ◽  
...  
Keyword(s):  
Cancer Pain ◽  
Noncoding Rna ◽  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Therapeutic Targets ◽  
Bone Cancer ◽  
Clinical Problem ◽  
Differentially Expressed ◽  
Bone Cancer Pain ◽  
Protein Protein Interaction

Bone cancer pain (BCP)–depression comorbidity has become a complex clinical problem during cancer treatment; however, its underlying molecular mechanisms have not been clarified. Several long noncoding RNAs (lncRNAs) have been demonstrated to be promising therapeutic targets in depression, but research on the role of lncRNAs in BCP–depression comorbidity has been limited. Therefore, high-throughput RNA sequencing was performed to detect differentially expressed profiles in the amygdala of a BCP–depression rat model in this study. We detected 330 differentially expressed mRNAs (DEmRNAs) and 78 differentially expressed lncRNAs (DElncRNAs) in the BCP–depression comorbidity model and then verified the expression of six DEmRNAs and six DElncRNAs with the greatest degrees of difference by RT-qPCR. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed that differentially expressed genes were strongly enriched in inflammatory and immunologic systemic responses. Then the nuclear factor kappa B (NF-κB) signaling pathway and the Th17 differentiation pathway showed significant differences, as determined by Western blot analysis. Finally, we constructed a protein–protein interaction (PPI) network to explore the potential regulatory mechanism of DEmRNAs. In conclusion, our study reveals a new resource for the understanding of dysregulated lncRNAs and mRNAs in BCP–depression comorbidity and provides novel potential therapeutic targets for further approaches.

scholarly journals circStrn3 is involved in bone cancer pain regulation in a rat model

2020 ◽  
Vol 52 (5) ◽  
pp. 495-505
Author(s):  
Yiwen Zhang ◽  
Xiaoxia Zhang ◽  
Zumin Xing ◽  
Shuyi Tang ◽  
Hanwen Chen ◽  
...  
Keyword(s):  
Cancer Pain ◽  
Rat Model ◽  
Molecular Mechanisms ◽  
Bone Cancer ◽  
Test Body ◽  
Differentially Expressed ◽  
Bone Cancer Pain ◽  
Biological Regulation ◽  
Cellular Processes ◽  
Simplex Infection

Abstract Bone cancer pain (BCP) is a common chronic pain that is caused by a primary or metastatic bone tumor. More detailed molecular mechanisms of BCP are warranted. In this study, we established a BCP rat model. The von Frey hair test, body weight, and hematoxylin and eosin staining were employed. We screened differentially expressed circRNAs (DECs) between the BCP group and sham group. The results revealed that 850 DECs were significantly up-regulated and 644 DECs were significantly down-regulated in the BCP group. Furthermore, we identified 1177 differentially expressed genes (DEGs) significantly up-regulated and 565 DEGs significantly down-regulated in the BCP group. Gene Ontology annotation of all 1742 DEGs revealed that biological regulation of metabolic processes, cellular processes, and binding were the top enriched terms. For Kyoto Encyclopedia of Genes and Genomes analysis, phagosome, HTLV-I infection, proteoglycans in cancer, and herpes simplex infection were significantly enriched in this study. In addition, we identified four selected circRNAs, chr6:72418120|72430205, chr20:7561057|7573740, chr18:69943105|69944476, and chr5:167516581|167558250, by quantitative real time PCR. chr6:72418120|72430205 (circStrn3) was selected for further study based on expression level and the circRNA–miRNA–mRNA network table. Western blot analysis suggested that knockdown of circStrn3 could effectively induce Walker 256 cell apoptosis. In summary, our study provided a more in-depth understanding of the molecular mechanisms of BCP.

scholarly journals Differential Proteomic Analysis of the Spinal Cord in Bone Cancer Pain Rats by Two-Dimensional Gel Electrophoresis

2021 ◽  
Author(s):  
Minjing Peng ◽  
Yanqiong Wu ◽  
Shanchun Su ◽  
Shengjun Wan ◽  
Lingyu Zhou ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Cancer Pain ◽  
Gel Electrophoresis ◽  
Bone Cancer ◽  
Bone Cancer Pain ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Altered Expression ◽  
Spectrometric Data ◽  
Underlying Mechanisms

Abstract Background: Bone cancer pain (BCP) is a common chronic pain that is caused by a primary or metastatic bone tumor. It is refractory to currently available clinical treatment owing to its complicated underlying mechanisms. Methods: In this study, we used proteomics approaches to investigate expressional changes of the rat spinal cord proteome from 7 to 21 d after inoculation. Proteins from the rat L4-6 spinal cord homogenates of BCP and Sham animals were fractionated by two-dimensional (2-DE) gel electrophoresis to produce a high-resolution map of the spinal cord soluble proteins. Proteins showing altered expression levels between BCP and Sham were selected. Results: A total of 60 spots were obtained, and isolated proteins were in-gel trypsin-digested and the resulting peptides were analyzed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. Using the mass spectrometric data, 34 differentially expressed proteins (DEPs) were identified. GO analysis of the identified proteins allowed us to explore the function of the represented proteins. Conclusions: Based on these results, the identified proteins may contribute to the maintenance of BCP, and may provided new or valuable information in the discovery of new therapeutic targets for BCP.

scholarly journals Bioinformatics Analysis of Circular RNA Expression Profiles in HBx-Transfected HepG2 Cells by Transcriptional Sequencing

2021 ◽  
Author(s):  
Luzheng Liu ◽  
Jiacheng Chen ◽  
Liang Chen ◽  
Cheng Chen ◽  
Dafeng Xu ◽  
...  
Keyword(s):  
Cell Lines ◽  
Hepg2 Cells ◽  
Target Genes ◽  
Expression Profiles ◽  
Circular Rna ◽  
Differentially Expressed ◽  
Circular Rnas ◽  
Bioinformatics Analyses ◽  
Go Analysis ◽  
Micro Rnas

Abstract BACKGROUND Circular RNA (CircRNA) and HBx genes separately play essential roles in the occurrence and development of hepatitis B (HBV)-related hepatocellular carcinoma (HCC). However, whether HBx expression in HCC is co-related to differential circRNA patterns remains unknown. METHODS HCC cell lines with HBx overexpression (HepG2 H6679) and empty vector control (HepG2 H5298) were successfully constructed. The high-throughput second-generation transcriptome sequencing technology (RNA-seq) was employed to sequence the two cell lines, and the selected circRNAs were verified by qPCR (quantitative real-time PCR). The differentially expressed circRNAs were analyzed. Bioinformatics analyses, including clustering, differential expression, GO analysis, and KEGG pathway, were performed. Target Scan and Miranda software were employed to predict miRNAs corresponding with circRNAs. RESULTS We identified 1120 circRNAs upregulated and 1447 circRNAs downregulated in HepG2 cell lines with HBx overexpression compared to its control. We selected 36 circRNAs with significant differences (also consistent with log2fold change absolute value ≥ 1.0 or P ≤ 0.05) displayed by cluster analysis and then performed qPCR validation. Among them, 15 circRNAs (hsa_circ_0005603, hsa_circ_0004448, hsa_circ_0006845, hsa_circ_0064654, hsa_circ_0006460, hsa_circ_0045350, hsa_circ_0000824, hsa_circ_0005227, hsa_circ_0067991, hsa_circ_0064656, hsa_circ_0005224, circRNA11716, circRNA759, circRNA14848 and circRNA13751) are consistent with sequencing results. Hsa_circ_0005603 and hsa_circ_0006845 showed significant differences and were chosen for further study. GO analysis shows that many target genes are involved in biological processes, cellular components, and molecular functions. Nearly 193 target genes were enriched on KEGG pathways analysis. Actin cytoskeleton regulation, tight junction, and FoxO signaling pathway are among the top three pathways involved in most genes. We predicted that hsa_circ_0005603 might interact with micro-RNAs, including miR-182-5p, hsa-miR-27a-3p, hsa-miR-98-5p, and hsa-miR-198, that might thereby regulate downstream genes involved in tumor progression. Similarly, hsa_circ_0006845 was predicted to be referred to HBV-related HCC by acting as a sponge for hsa-miR-106a-3p and hsa-miR-198. Furthermore, we discovered two novel circular RNAs (circRNA11716 and circRNA13751) which might be involved in HCC occurrence. CONCLUSION In this study, we comprehensively explored the differentially expressed circRNAs in HepG2 cells with different HBx expression, and our results indicate that hsa_circ_0005603, hsa_circ_0006845, and novel circular RNAs (circRNA11716 and circRNA13751) might play an important role in HBV-related HCC, deserving further research.

scholarly journals MiR-135-5p alleviates bone cancer pain by regulating astrocyte-mediated neuroinflammation in spinal cord through JAK2/STAT3 signaling pathway

2021 ◽  
Author(s):  
Ming Liu ◽  
Xuefeng Cheng ◽  
Hong Yan ◽  
Jingli Chen ◽  
Caihua Liu ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Cancer Pain ◽  
Signaling Pathway ◽  
Expression Profiles ◽  
Bone Cancer ◽  
Bone Cancer Pain ◽  
Luciferase Reporter ◽  
Stat3 Signaling ◽  
Stat3 Signaling Pathway ◽  
Gene Assay

Bone cancer pain (BCP) was associated with microRNA dysregulation. In this study, we intended to clarify the potential role of miR-135-5p in a BCP mouse model, which was established by tumor cell implantation (TCI) in the medullary cavity of the mouse femur. The BCP related behaviors were tested, including the paw withdrawal mechanical threshold (PWMT) and number of spontaneous flinches (NSF). The miRNA expression profiles in astrocytes of the sham and tumor groups were compared, and miRNA microarray and quantitative real-time PCR (qRT-PCR) assays confirmed that the amount of expression of miR-135-5p was significantly decreased in astrocytes of the tumor group. Gain- and loss-of-function studies showed that miR-135-5p could inhibit astrocytes activation and inflammation cytokines (TNF-α and IL-1β) expression. The relation between miR-135-5p and JAK2 was detected by bioinformatic analysis and dual luciferase reporter gene assay. By conducting in vitro experiments, it was shown that the miR-135-5P mimics lowered the level of JAK2/STAT3 proteins and inflammatory factors in astrocytes. Moreover, in vivo analysis on BCP mice model indicated that the miR-135-5p agonist could sufficiently increase PWMT and decrease NSF. Meanwhile, reduced activation of astrocytes in the spinal cord, as well as decreased expression of JAK2/STAT3 and inflammatory mediators, were found after miR-135-5p agonist treatment. Collectively, the results showed that miR-135-5p could potentially reduce BCP in mice through inhibiting astrocyte-mediated neuroinflammation and blocking of the JAK2/STAT3 signaling pathway, indicating that the upregulation of miR-135-5P could be a therapeutic focus in BCP treatment.

Genome-Wide Identification of differently expressed lncRNAs, mRNAs, and circRNAs in patients with osteoarthritis

2020 ◽  
Vol 15 ◽  
Author(s):  
Yeqing Sun ◽  
Lei Chen ◽  
Yingqi Zhang ◽  
Jincheng Zhang ◽  
Shashi Ranjan Tiwari
Keyword(s):  
Regulatory Networks ◽  
Expression Profiles ◽  
Interleukin 1 ◽  
Interaction Network ◽  
Circular Rna ◽  
Negative Regulation ◽  
Differentially Expressed ◽  
Positive Regulation ◽  
Proteinprotein Interaction ◽  
Differently Expressed Genes

Background: Osteoarthritis (OA), one of the most important causes leading to joint disability, was considered as an untreatable disease. A series of genes were reported to regulate the pathogenesis of OA, including microRNAs, Long non-coding RNAs and Circular RNA. So far, the expression profiles and functions of lncRNAs, mRNAs, and circRNAs in OA are not fully understood. Objective: The present study aimed to identify differently expressed genes in OA. Methods: The present study conducted RNA-seq to identify differently expressed genes in OA. Ontology (GO) analysis was used to analysis the Molecular Function and Biological Process. KEGG pathway analysis was used to perform the differentially expressed lncRNAs in biological pathways. Results: Hierarchical clustering revealed a total of 943 mRNAs, 518 lncRNAs, and 300 circRNAs were dysregulated in OA compared to normal samples. Furthermore, we constructed differentially expressed mRNAs mediated proteinprotein interaction network, differentially expressed lncRNAs mediated trans regulatory networks, and competitive endogenous RNA (ceRNA) to reveal the interaction among these genes in OA. Bioinformatics analysis revealed these dysregulated genes were involved in regulating multiple biological processes, such as wound healing, negative regulation of ossification, sister chromatid cohesion, positive regulation of interleukin-1 alpha production, sodium ion transmembrane transport, positive regulation of cell migration, and negative regulation of inflammatory response. To the best of our knowledge, this study for the first time revealed the expression pattern of mRNAs, lncRNAs and circRNAs in OA. Conclusion: This study provided novel information to validate these differentially expressed RNAs may be as possible biomarkers and targets in OA.

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