scholarly journals Downregulated MiR-206 expression promotes the proliferation and migration of macrophages by regulating IL-17A/REG3A pathway

2020 ◽  
Vol 18 ◽  
pp. 205873922091749
Author(s):  
Xuan Huang ◽  
Fang Gong ◽  
Zhixian Lu ◽  
Jie Zhu ◽  
Qun Yu
Keyword(s):  
Pcr Analysis ◽  
Chain Reaction ◽  
Inflammatory Infiltration ◽  
Interleukin 17A ◽  
Polymerase Chain ◽  
And Migration ◽  
Cell Migration And Proliferation ◽  
Migration Of Macrophages ◽  
Proliferation And Migration

This study sought to investigate the role of miR-206 in polymyositis/dermatomyositis (PM/DM) development. Transwell and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay were performed to investigate cell migration and proliferation. Real-time polymerase chain reaction (PCR) analysis was used to determine the expression of miR-206, interleukin-17A (IL-17A), IL-17 receptor A (IL-17RA), and regenerating islet-derived protein 3-alpha (REG3A). Significantly, miR-206 mimics decreased macrophage migration and proliferation, while miR-206 inhibitors exhibited the opposite effects. Indeed, elevating IL-17RA levels resulted in increased REG3A expression, which was inhibited by IL-17RA siRNA. Besides, miR-206 mimics decreased IL-17A and REG3A expressions, but miR-206 inhibitors showed opposite effects. Moreover, miR-206 expression in PM/DM patients was significantly reduced compared with the healthy controls, while IL-17A and REG3A expressions substantially increased among PM/DM patients. These findings suggested that downregulation of miR-206 increased the migration and proliferation of macrophages via IL-17A/REG3A signaling pathway, which could promote the inflammatory infiltration in PM/DM.

scholarly journals Effect of Lentivirus-Mediated miR-499a-3p on Human Umbilical Vein Endothelial Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-13
Author(s):  
Huilei Zheng ◽  
Juan Li ◽  
Ying Chen ◽  
Danping Gong ◽  
Jianlin Wen ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Human Umbilical Vein ◽  
Polymerase Chain ◽  
And Migration ◽  
Umbilical Vein Endothelial Cells ◽  
Cell Migration And Proliferation ◽  
Scratch Tests ◽  
Proliferation And Migration

Objective. To explore the possible role of miR-499a-3p in the function of primary human umbilical vein endothelial cells (HUVECs) and the expression of ADAM10 in primary HUVEC. Method. miR-499a-3p was first transfected into primary HUVECs via lentivirus vector. The viability, proliferation, and migration of stable transfected primary HUVEC were then determined by flow cytometry, CCK8 assays, scratch tests, and Transwell tests. The transcription of miR-499a-3p and ADAM10 was examined by reverse transcription-polymerase chain reaction (RT-PCR), and the expression of ADAM10 was examined by Western blot (WB). Results. After transfection, miR-499a-3p transcription was significantly increased (P<0.01), compared to the blank and nonspecific control (NC) groups, while both ADAM10 transcription and expression were significantly decreased (P<0.05). In contrast, in the inhibitors group, miR-499a-3p transcription was significantly reduced (P<0.05) whereas both ADAM10 transcription and expression were significantly increased (P<0.05). The viability, proliferation, and migration of primary HUVECs were significantly impaired (P<0.05) by the transfection of miR-499a-3p but enhanced by miR-499a-3p inhibitors (P<0.05). Conclusions. Upregulation of miR-499a-3p transcription will inhibit the expression of ADAM10 in HUVECs; cell migration and proliferation, however, promote apoptosis. And reverse effects were established by downregulation of miR-499a-3p transcription. All these effects may be achieved by regulating the transcription and expression of ADAM10. These results combined suggested that miR-499a-3p may affect the proliferation, migration, and apoptosis of endothelial cells and regulate AS by regulating ADAM10. miR-499a-3p may become a candidate biomarker for the diagnosis of unstable angina pectoris (UA).

scholarly journals Inhibition of miR-155 attenuates abdominal aortic aneurysm in mice by regulating macrophage-mediated inflammation

2018 ◽  
Vol 38 (3) ◽  
Author(s):  
Zhidong Zhang ◽  
Kai Liang ◽  
Gangqiang Zou ◽  
Xiaosan Chen ◽  
Shuaitao Shi ◽  
...  
Keyword(s):  
Opposite Effect ◽  
Abdominal Aortic Aneurysms ◽  
Aortic Aneurysms ◽  
Chain Reaction ◽  
Monocyte Chemoattractant ◽  
Qrt Pcr ◽  
Abdominal Aortic ◽  
Polymerase Chain ◽  
And Migration ◽  
Proliferation And Migration

The aim of the present study was to identify abdominal aortic aneurysms (AAA)-associated miR-155 contributing to AAA pathology by regulating macrophage-mediated inflammation. Angiotensin II (AngII)–infused apolipoprotein E-deficient (ApoE-/-) mice and THP-1 cells model of miR-155 overexpression and deficiency were used in the experiments. The expression of miR-155 was detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Cytokines were evaluated using enzyme-linked immunoabsorbent assay (ELISA). Western blotting was used to measure the levels of MMP-2, MMP-9, iNOS, and monocyte chemoattractant protein (MCP)-1 proteins. Immunostaining and transwell were used to determine CD68, elastic collagen, proliferation, and migration of vascular smooth muscle cells (VSMCs). The results showed that miR-155 and cytokines were up-regulated in AAA patients or ApoE-/- mice. Overexpression of miR-155 enhanced MMP-2, MMP-9, iNOS, and MCP-1 levels, and stimulated the proliferation and migration of VSMCs. Meanwhile, inhibition of miR-155 had the opposite effect. In addition, histology demonstrated accumulation of CD68 and elastic collagen-positive areas significantly decreased in miR-155 antagomir injection group. In conclusion, the results of the present study suggest that inhibiting miR-155 is crucial to prevent the development of AAA by regulating macrophage inflammation.

scholarly journals Na+/Ca2+ exchangers regulate the migration and proliferation of human gastric myofibroblasts

2013 ◽  
Vol 305 (8) ◽  
pp. G552-G563 ◽  
Author(s):  
Lajos V. Kemény ◽  
Andrea Schnúr ◽  
Mátyás Czepán ◽  
Zoltán Rakonczay ◽  
Eleonóra Gál ◽  
...  
Keyword(s):  
Tumor Development ◽  
Oscillatory Activity ◽  
Pcr Analysis ◽  
Gastric Diseases ◽  
Protein Levels ◽  
Intracellular Ca ◽  
Transitional Cells ◽  
And Migration ◽  
Proliferation And Migration

Gastrointestinal myofibroblasts are contractile, electrically nonexcitable, transitional cells that play a role in extracellular matrix production, in ulcer healing, and in pathophysiological conditions they contribute to chronic inflammation and tumor development. Na+/Ca2+ exchangers (NCX) are known to have a crucial role in Ca2+ homeostasis of contractile cells, however, no information is available concerning the role of NCX in the proliferation and migration of gastrointestinal myofibroblasts. In this study, our aim was to investigate the role of NCX in the Ca2+ homeostasis, migration, and proliferation of human gastrointestinal myofibroblasts, focusing on human gastric myofibroblasts (HGMs). We used microfluorometric measurements to investigate the intracellular Ca2+ and Na+ concentrations, PCR analysis and immunostaining to show the presence of the NCX, patch clamp for measuring NCX activity, and proliferation and migration assays to investigate the functional role of the exchanger. We showed that 53.0 ± 8.1% of the HGMs present Ca2+ oscillations, which depend on extracellular Ca2+ and Na+, and can be inhibited by NCX inhibitors. NCX1, NCX2, and NCX3 were expressed at both mRNA and protein levels in HGMs, and they contribute to the intracellular Ca2+ and Na+ homeostasis as well, regardless of the oscillatory activity. NCX inhibitors significantly blocked the basal and insulin-like growth factor II-stimulated migration and proliferation rates of HGMs. In conclusion, we showed that NCX plays a pivotal role in regulating the Ca2+ homeostasis, migration, and proliferation of HGMs. The inhibition of NCX activity may be a potential therapeutic target in hyperproliferative gastric diseases.

Role of LncRNA CASC9 in oral carcinoma and its downstream gene expression and mechanism detected by nanomagnetic bead-based polymerase chain reaction

2021 ◽  
Vol 11 (7) ◽  
pp. 1017-1023
Author(s):  
Junjie Liu ◽  
Hong Mu ◽  
Xiangyong Cheng ◽  
Gangying Yuan ◽  
Qinqin Wang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Oral Carcinoma ◽  
Luciferase Reporter ◽  
Chain Reaction ◽  
Total Rna ◽  
Polymerase Chain ◽  
And Migration ◽  
The Impact ◽  
Downstream Gene Expression

In view of the increasing incidence of oral carcinoma (OC), an in-depth understanding of the mechanism is required for future prevention and treatment. In modern medical research, the etiology of tumor diseases is mainly focused on gene changes, among which lncRNA is a trending topic. We speculated that lncRNA CASC9 may attribute to OC occurrence. To verify our conjecture, we extracted the total RNA of the research samples through nanomagnetic beads, detected the expression of CASC9 downstream genes miR-383-5p and KIF3B by polymerase chain reaction (PCR), and analyzed the impact of the three on OC cells and the targeting relationship. The total RNA extracted by nanomagnetic beads were found to be able to effectively improve the PCR speed, yield and the accuracy of the results, and reduce the allergic amplification results of the samples to be tested to obtain the best results. The detection result showed high CASC9 and KIF3B expression, and low miR-383-5p expression in OC cells. Inhibition of CASC9 and KIF3B and elevation of miR-383-5p can inhibit the viability of OC cells and boost apoptosis. Through dual-luciferase reporter (DLR) assay, the targeted regulation relationship between CASC9 and miR-383-5p and between miR-383-5p and KIF3B, was determined. Rescue experiments confirmed that CASC9 was able to modulate OC cell biological behaviors by targeting the miR-383-5p/KIF3B axis. Therefore, we argue that with high expression in OC, CASC9 can enhance the proliferation and migration of OC cells and inhibit the apoptosis through targeting the miR-383-5p/KIF3B axis.

scholarly journals MicroRNA-197 regulates chondrocyte proliferation, migration, and inflammation in pathogenesis of osteoarthritis by targeting EIF4G2

2020 ◽  
Vol 40 (9) ◽  
Author(s):  
Shijie Gao ◽  
Liang Liu ◽  
Shibo Zhu ◽  
Dawei Wang ◽  
Qiang Wu ◽  
...  
Keyword(s):  
Target Gene ◽  
Inverse Correlation ◽  
Luciferase Reporter ◽  
Pcr Analysis ◽  
Chondrocyte Proliferation ◽  
Normal Cartilage ◽  
Tnf Α ◽  
And Migration ◽  
Proliferation And Migration

Abstract Recent studies have demonstrated that microRNAs (miRNAs) are involved in many pathological conditions including osteoarthritis (OA). In the present study, we aimed to investigate the role of miR-197 in OA and the potential molecular mechanism. The expression levels of miR-197 were detected by quantitative real-time PCR analysis. Cell proliferation and migration abilities were performed by 3-(4,5-dimethylthiazol-2-yl)-2,5-di-phenyltetrazolium bromide and transwell assays. The concentrations of inflammatory cytokines, including IL-1β, IL-6, and TNF-α, were detect using ELISA assay. Furthermore, luciferase reporter and rescue assays were applied to identify the functional target gene of miR-197 in OA. The results showed that miR-197 expression was significantly down-regulated in the OA cartilage tissues compared with normal cartilage tissues, accompanied by up-regulation of EIF4G2 expression. An inverse correlation was found between EIF4G2 and miR-197 expressions in OA cartilage tissues. Treatment with miR-197 mimics promoted the growth and migration abilities of chondrocytes, while miR-197 inhibitors induced the opposite effects. Furthermore, restoration of miR-197 significantly decreased IL-1β, IL-6, and TNF-α expression, whereas knockdown of miR-197 led to a induction in these inflammatory mediators. Moreover, EIF4G2 was predicted and confirmed as a directly target of miR-197. Overexpressed miR-197 could down-regulate EIF4G2 expression in chondrocytes, while miR-197 knockdown could elevate EIF4G2 expression. Additionally, EIF4G2 overexpression reversed the effects of miR-197 mimics on chondrocytes proliferation, migration, and inflammation. Taken together, our study demonstrated that miR-197 promotes chondrocyte proliferation, increases migration, and inhibits inflammation in the pathogenesis of OA by targeting EIF4G2, indicating the potential therapeutic targets of the miR-197/EIF4G2 axis for OA treatment.

Role of Anaerobes in Chronic Sinusitis: Will Polymerase Chain Reaction Solve the Debate

2002 ◽  
Vol 127 (5) ◽  
pp. 384-386 ◽  
Author(s):  
Hassan H. Ramadan ◽  
Peter H. Mathers ◽  
Heather Schwartzbauer
Keyword(s):  
Chronic Rhinosinusitis ◽  
Chronic Sinusitis ◽  
Anaerobic Bacteria ◽  
Sinus Surgery ◽  
Pcr Analysis ◽  
Chain Reaction ◽  
Culture Techniques ◽  
Standard Culture ◽  
Polymerase Chain

BACKGROUND: Bacteriology of chronic sinusitis continues to be a matter of debate, particularly the role of anaerobes. Some authors suggest that anaerobes play a significant role whereas others suggest a minimal role. Those who suggest a significant role argue that standard culture techniques are the culprits. OBJECTIVE: The purpose of this study was to use polymerase chain reaction (PCR) on sinus specimens for the presence of anaerobes and to compare them with standard culture techniques. METHODS: Sixty-four samples were obtained in a sterile fashion during sinus surgery and were sent for standard anaerobic cultures as well as anaerobic PCR analysis. RESULTS: Anaerobic bacteria were demonstrated in 5% of culture specimens, which is similar to recently published data. PCR identified anaerobic bacteria in 19% of the specimens ( P = 0.01) CONCLUSION: PCR analysis of surgical samples obtained during endoscopic sinus surgery for chronic sinusitis identified a significantly higher incidence of anaerobes (x4) compared with standard anaerobic culture technique. Chronic rhinosinusitis is one of the most common chronic diseases that affects individuals in the United States. It is estimated that >25 million office visits are made for sinusitis. 1 The financial impact of chronic sinusitis includes not only the direct medical costs of treatment but also the millions of hours of lost productivity caused by the disease. 1 Chronic rhinosinusitis is defined as the signs and symptoms of sinus inflammation that last longer than 12 weeks associated with documented sinus inflammation with imaging techniques ≥4 weeks after appropriate antibiotic therapy. 2 There is agreement in the literature regarding the bacterial etiology of acute rhinosinusitis. However, the etiology of chronic rhinosinusitis is still unclear despite numerous articles about the bacteriology of chronic rhinosinusitis. 3–10 The role of aerobes is more clear than the role of anaerobes in chronic rhinosinusitis; many conflicting reports have been published about the role of anaerobes as etiologic factors in chronic rhinosinusitis. 3–10 The incidence of anaerobes obtained on standard cultures from patients with chronic rhinosinusitis ranges from 0% to 56%. 3–10 The reports that show a high yield of anaerobes argue that the technique used to obtain the specimens, the method of transportation, and even specimen collection are reasons why other reports did not yield high levels of anaerobes. 4–5 With increasing numbers of patients with chronic rhinosinusitis and with increasing bacterial resistance to antimicrobials, which is blamed on the improper use of antimicrobials, knowledge of the bacteriology becomes important in the treatment. The role of aerobes was addressed in a prior publication 11 ; the role of anaerobic bacteria as an etiologic cause of chronic rhinosinusitis is addressed in this report. The goal of this study was to test specimens, using 2 different techniques, obtained during endoscopic sinus surgery from individuals who had chronic rhinosinusitis.

scholarly journals Toxoplasma gondii Cyclophilin 18 Regulates the Proliferation and Migration of Murine Macrophages and Spleen Cells

2010 ◽  
Vol 17 (9) ◽  
pp. 1322-1329 ◽  
Author(s):  
Hany M. Ibrahim ◽  
Xuenan Xuan ◽  
Yoshifumi Nishikawa
Keyword(s):  
Toxoplasma Gondii ◽  
Cell Types ◽  
The Body ◽  
Host Cells ◽  
High Dose ◽  
Spleen Cells ◽  
And Migration ◽  
Migration Of Macrophages ◽  
Proliferation And Migration

ABSTRACT Toxoplasma gondii is an intracellular parasite that shows a unique capacity to infect a variety of cell types in warm-blooded animals. It can invade and survive well inside immune cells, such as macrophages, that disseminate the parasite around the body because of their migratory properties. The aim of the present study was to evaluate the role of T. gondii cyclophilin 18 (TgCyp18) in the proliferation and migration of macrophages and spleen cells (mainly T lymphocytes) in order to understand the effects of TgCyp18 on the dynamics of the infection. A high dose of TgCyp18 enhanced the proliferation of macrophages and spleen cells in a cysteine-cysteine chemokine receptor 5 (CCR5)-independent way. In contrast, TgCyp18 controlled the migration of macrophages and spleen cells in dose- and CCR5-dependent manners. Our data suggest that TgCyp18 recruits cells and enhances the growth of host cells at the site of infection for maintenance of the interaction between the parasite and host.

Role of microenvironment on proliferation and migration of an SDHB silenced murine Pheochromocytoma cell line

2017 ◽  
Author(s):  
Serena Martinelli ◽  
Vanessa D'Antongiovanni ◽  
Susan Richter ◽  
Letizia Canu ◽  
Tonino Ercolino ◽  
...  
Keyword(s):  
Cell Line ◽  
Pheochromocytoma Cell ◽  
And Migration ◽  
Proliferation And Migration

Exosomal miR-214-5p Released from Glioblastoma Cells Modulates Inflammatory Response of Microglia after Lipopolysaccharide Stimulation through Targeting CXCR5

2019 ◽  
Vol 18 (1) ◽  
pp. 78-87 ◽  
Author(s):  
Jian-kai Yang ◽  
Hong-jiang Liu ◽  
Yuanyu Wang ◽  
Chen Li ◽  
Ji-peng Yang ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Critical Role ◽  
Luciferase Reporter ◽  
Clinical Prognosis ◽  
Direct Target ◽  
And Migration ◽  
High Level ◽  
Cxcr5 Expression ◽  
Proliferation And Migration

Background and Objective: Exosomes communicate inter-cellularly and miRNAs play critical roles in this scenario. MiR-214-5p was implicated in multiple tumors with diverse functions uncovered. However, whether miR-214-5p is mechanistically involved in glioblastoma, especially via exosomal pathway, is still elusive. Here we sought to comprehensively address the critical role of exosomal miR-214-5p in glioblastoma (GBM) microenvironment.Methods:The relative expression of miR-214-5p was determined by real-time PCR. Cell viability and migration were measured by MTT and transwell chamber assays, respectively. The secretory cytokines were measured with ELISA kits. The regulatory effect of miR-214-5p on CXCR5 expression was interrogated by luciferase reporter assay. Protein level was analyzed by Western blot.Results:We demonstrated that miR-214-5p was aberrantly overexpressed in GBM and associated with poorer clinical prognosis. High level of miR-214-5p significantly contributed to cell proliferation and migration. GBM-derived exosomal miR-214-5p promoted inflammatory response in primary microglia upon lipopolysaccharide challenge. We further identified CXCR5 as the direct target of miR-214- 5p in this setting.Conclusion:Overexpression of miR-214-5p in GBM modulated the inflammatory response in microglia via exosomal transfer.

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