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Circular RNAs (circRNAs) are key regulators of cervical cancer (CC) progression. This study aimed to elucidate the role and mechanism of circ_0051428, a novel circRNA, in CC tumorigenesis. Quantitative real-time polymerase chain reaction and western blotting analyses confirmed that circ_0051428 and matrix metalloprotein-2 (MMP2) were overexpressed in CC, whereas the microRNA miR-885-3p was poorly expressed. After performing a series of in vitro and in vivo experiments, circ_0051428 knockdown was shown to repress CC cell invasion and proliferation in vitro, and hamper tumor formation in vivo. Dual-luciferase reporter and RNA-binding protein immunoprecipitation experiments verified that circ_0051428 interacts with miR-885-3p to regulate the target gene MMP2 of miR-885-3p in CC. In addition, miR-885-3p knockdown offset the anticancer effects of circ_0051428 or MMP2 knockdown on CC cell malignancy. Overall, this study revealed that circ_0051428 executes a tumor-promoting function in CC pathogenesis by modulating the miR-885-3p/MMP2 axis. Our findings provide a novel approach for CC treatment.

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.

Graves ophthalmopathy (GO), a manifestation of Graves' disease, is an organ-specific autoimmune disease. Intravenous glucocorticoid therapy (ivGCs) is the first-line treatment for moderate-to-severe and active GO. However, ivGCs is only effective in 70%–80% of GO patients. Insensitive patients who choose 12-week ivGCs not only were delayed in treatment but also took the risk of adverse reactions of glucocorticoids. At present, there is still a lack of effective indicators to predict the therapeutic effect of ivGCs. Therefore, the purpose of this study is to find biomarkers that can determine the sensitivity of ivGCs before the formulation of treatment, and to clarify the mechanism of its regulation of ivGCs sensitivity. This study first characterized the miRNA profiles of plasma exosomes by miRNA sequencing to identify miRNAs differentially expressed between GO patients with significant improvement (SI) and non-significant improvement (NSI) after ivGCs treatment. Subsequently, we analyzed the function of the predicted target genes of differential miRNAs. According to the function of the target genes, we screened 10 differentially expressed miRNAs. An expanded cohort verification showed that compared with NSI patients, mir-885-3p was upregulated and mir-4474-3p and mir-615-3p were downregulated in the exosomes of SI patients. Based on statistical difference and miRNA function, mir-885-3p was selected for follow-up study. The in vitro functional analysis of exosomes mir-885-3p showed that exosomes from SI patients (SI-exo) could transfer mir-885-3p to orbital fibroblasts (OFs), upregulate the GRE luciferase reporter gene plasmid activity and the level of glucocorticoid receptor (GR), downregulate the level of inflammatory factors, and improve the glucocorticoid sensitivity of OFs. Moreover, these effects can be inhibited by the corresponding miR inhibitor. In addition, we found that high levels of mir-885-3p could inhibit the AKT/NFκB signaling pathway, upregulate the GRE plasmid activity and GR level, and downregulate the level of inflammatory factors of OFs. Moreover, the improvement of glucocorticoid sensitivity by mir-885-3p transmitted by SI-exo can also be inhibited by the AKT/NFκB agonist. Finally, through the in vivo experiment of the GO mouse model, we further determined the relationship between exosomes' mir-885-3p sequence, AKT/NFκB signaling pathway, and glucocorticoid sensitivity. As a conclusion, plasma exosomes deliver mir-885-3p and inhibit the AKT/NFκB signaling pathway to improve the glucocorticoid sensitivity of OFs. Exosome mir-885-3p can be used as a biomarker to determine the sensitivity of ivGCs in GO patients. [ABSTRACT FROM AUTHOR]

Graves ophthalmopathy (GO), a manifestation of Graves’ disease, is an organ-specific autoimmune disease. Intravenous glucocorticoid therapy (ivGCs) is the first-line treatment for moderate-to-severe and active GO. However, ivGCs is only effective in 70%–80% of GO patients. Insensitive patients who choose 12-week ivGCs not only were delayed in treatment but also took the risk of adverse reactions of glucocorticoids. At present, there is still a lack of effective indicators to predict the therapeutic effect of ivGCs. Therefore, the purpose of this study is to find biomarkers that can determine the sensitivity of ivGCs before the formulation of treatment, and to clarify the mechanism of its regulation of ivGCs sensitivity. This study first characterized the miRNA profiles of plasma exosomes by miRNA sequencing to identify miRNAs differentially expressed between GO patients with significant improvement (SI) and non-significant improvement (NSI) after ivGCs treatment. Subsequently, we analyzed the function of the predicted target genes of differential miRNAs. According to the function of the target genes, we screened 10 differentially expressed miRNAs. An expanded cohort verification showed that compared with NSI patients, mir-885-3p was upregulated and mir-4474-3p and mir-615-3p were downregulated in the exosomes of SI patients. Based on statistical difference and miRNA function, mir-885-3p was selected for follow-up study. The in vitro functional analysis of exosomes mir-885-3p showed that exosomes from SI patients (SI-exo) could transfer mir-885-3p to orbital fibroblasts (OFs), upregulate the GRE luciferase reporter gene plasmid activity and the level of glucocorticoid receptor (GR), downregulate the level of inflammatory factors, and improve the glucocorticoid sensitivity of OFs. Moreover, these effects can be inhibited by the corresponding miR inhibitor. In addition, we found that high levels of mir-885-3p could inhibit the AKT/NFκB signaling pathway, upregulate the GRE plasmid activity and GR level, and downregulate the level of inflammatory factors of OFs. Moreover, the improvement of glucocorticoid sensitivity by mir-885-3p transmitted by SI-exo can also be inhibited by the AKT/NFκB agonist. Finally, through the in vivo experiment of the GO mouse model, we further determined the relationship between exosomes’ mir-885-3p sequence, AKT/NFκB signaling pathway, and glucocorticoid sensitivity. As a conclusion, plasma exosomes deliver mir-885-3p and inhibit the AKT/NFκB signaling pathway to improve the glucocorticoid sensitivity of OFs. Exosome mir-885-3p can be used as a biomarker to determine the sensitivity of ivGCs in GO patients.

Trastuzumab has been applied widely in the treatment of breast cancer. The majority of initial responders display disease progression again within one year. Regardless of the high resistance rate, the molecular mechanisms affected this disease are not well understood. MicroRNAs are small, non-coding RNA molecules that involved in gene regulation. There is evidence that promotes miRNAs as potential candidates to mediate therapeutic actions by targeting genes involved in drug response. The purpose of this study is to evaluate miR-885-3p in HER2 positive breast cancer chemoresistance. Trastuzumab-resistant BT-474 cells were generated by in vitro culture of BT-474 cells continuously in the presence of trastuzumab about 9 months. The relative expression of miR-885-3p to U6 RNA was evaluated in trastuzumab-resistant and sensitive cells using Relative Real-Time PCR. The Mann-Whitney test is used to compare the differences between the two groups. The MTT assay showed that BT-474 breast cancer cells were resistant to this drug under long-term culturing with trastuzumab (p < 0.05). MiR-885-3p expression was also significantly downregulated in trastuzumab-resistant cells in comparison with the parent cells (p < 0.05).: As the relative expression of candidate microRNAs was statistically different in trastuzumab-resistant and sensitive cells, we hypothesize that miR-885-3p downregulation as a possible mechanism of trastuzumab resistance.

Background/Aims: Accumulative evidence has shown that miR-885-3p plays a crucial role in human carcinogenesis. From miRNA database, we also found that rs739837 polymorphism in miR-885-3p binding site within 3’-untranslated region of Vitamin D receptor (VDR), compromising the suppressive effect of miR-885-3p on VDR. Moreover, Vitamin D is involved in controlling the cell immune response and may play a role in pressure ulcers development. However, whether this polymorphism is actually linked with pressure ulcers remains unclear. The aim of the present study was to investigate the potential association between the rs739837 polymorphism and pressure ulcers, and to explore molecular mechanism of VDR in pressure ulcers. Methods: Luciferase assays were performed to validate the relationship between miR-885 and VDR, which was confirmed by western-blotting analysis. The relationship between rs739837 and the risk of pressure ulcers was explored using logistic regression analysis. Results: We first conducted statistical analysis to explore the association between the rs739837 genotype and risk of pressure ulcers, and found that the polymorphism genotype was significantly associated with the risk of pressure ulcers (OR 0.68, 95% CI 0.43-0.95, P value = 0.02). We then searched the miRNA database online and identified VDR as a direct target. We established the negative regulatory relationship between miR-885-3p and VDR using luciferase assays. Meanwhile, we transfected cells with scramble control, miR-885-3p mimics, VDR siRNA and miR-885-3p inhibitors. The results further confirmed the negative regulatory relationship between miR-885-3p and VDR. Conclusion: VDR is a virtual target of miR-885-3p, and rs739837 might be a predictive biomarker for the risk of pressure ulcers.

Long non-coding RNAs (lncRNAs) have been proven to play important roles in carcinogenesis and development of numerous cancers, but their biological functions in glioblastoma remain largely unknown. In this study, we found that HOXB-AS1 was highly expressed in human glioblastoma tissues and cell lines, and was associated with survival time of patients. Further analysis showed that knock-down of HOXB-AS1 inhibited cell proliferation via inducing S phase cell cycle arrest and suppressed migration and invasion ability of cells. Mechanism study revealed that HOXB-AS1 is mainly located in cytoplasm and functions as competing endogenous RNA via sponging of miR-885-3p. Moreover, inhibition of miR-885-3p antagonized the effects of HOXB-AS1 knock-down and promoted proliferation, migration and invasion of glioblastoma cells. Finally, we found that sponging of miR-885-3p by HOXB-AS1 could further affect the expression of HOXB2. Taken together, we demonstrate that HOXB-AS1/miR-885-3p/HOXB2 axis regulates proliferation, migration and invasion of glioblastoma cells and can serve as a potential biomarker for the malignancy. [ABSTRACT FROM AUTHOR]

Background/Aims: Accumulative evidence has shown that miR-885-3p plays a crucial role in human carcinogenesis. From miRNA database, we also found that rs739837 polymorphism in miR-885-3p binding site within 3'-untranslated region of Vitamin D receptor (VDR), compromising the suppressive effect of miR-885-3p on VDR. Moreover, Vitamin D is involved in controlling the cell immune response and may play a role in pressure ulcers development. However, whether this polymorphism is actually linked with pressure ulcers remains unclear. The aim of the present study was to investigate the potential association between the rs739837 polymorphism and pressure ulcers, and to explore molecular mechanism of VDR in pressure ulcers. Methods: Luciferase assays were performed to validate the relationship between miR-885 and VDR, which was confirmed by western-blotting analysis. The relationship between rs739837 and the risk of pressure ulcers was explored using logistic regression analysis. Results: We first conducted statistical analysis to explore the association between the rs739837 genotype and risk of pressure ulcers, and found that the polymorphism genotype was significantly associated with the risk of pressure ulcers (OR 0.68, 95% CI 0.43-0.95, P value = 0.02). We then searched the miRNA database online and identified VDR as a direct target. We established the negative regulatory relationship between miR-885-3p and VDR using luciferase assays. Meanwhile, we transfected cells with scramble control, miR-885-3p mimics, VDR siRNA and miR-885-3p inhibitors. The results further confirmed the negative regulatory relationship between miR-885-3p and VDR. Conclusion: VDR is a virtual target of miR-885-3p, and rs739837 might be a predictive biomarker for the risk of pressure ulcers. [ABSTRACT FROM AUTHOR]

Breast cancer is the most common cancer in women worldwide. Numerous reports have demonstrated that circRNAs play an essential role in regulating the biological characteristics of breast cancer. However, there are currently no reports regarding the role of hsa&#95;circ&#95;0006014 in breast cancer. In this study, qRT-PCR was used to detect the expression of hsa&#95;circ&#95;0006014 and related genes. MTT, colony formation and Transwell assays were used to explore the potential biological functions of hsa&#95;circ&#95;0006014 in breast cancer cells. Western blotting was used to explore the potential molecular mechanisms involving hsa&#95;circ&#95;0006014. In vivo experiments were used to evaluate the influence of hsa&#95;circ&#95;0006014 on animal tumors. In this study, we found higher expression of hsa&#95;circ&#95;0006014 in breast tumor samples than in matched adjacent normal samples, and its expression was positively correlated with histological grade (grade iii). Phenotypically, hsa&#95;circ&#95;0006014 promoted the proliferation of MDA-MB-231 and MCF-7 breast cancer cells. Mechanistically, there were confirmed binding sites between hsa&#95;circ&#95;0006014 and miR-885-3p, and hsa&#95;circ&#95;0006014 promoted breast cancer cell proliferation partially by sponging miR-885-3p and influenced CDK2/CCNE1 and CDK4/6/CCND1. Furthermore, we found that hsa&#95;circ&#95;0006014 regulated NTRK2 through miR-885-3p to modulate the PIK3/AKT signaling pathway. Our results demonstrated that hsa&#95;circ&#95;0006014 promotes breast cancer progression by sponging miR-885-3p to regulate the NTRK2/PIK3CA/AKT axis.

Airway inflammation is one of the typical pathological characteristics of asthma. MicroRNAs (miRNAs) play important roles in regulating inflammation. Nevertheless, miRNA-885-3p (miR-885-3p)'s role in asthmatic inflammation and the underlying mechanism need to be explained. In this work, miR-885-3p expression and toll-like receptor 4 (TLR4) expression in asthma patients' plasma and lipopolysaccharide (LPS)-treated 16HBE cells were detected through quantitative real-time PCR. The interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels in 16HBE cell supernatant were examined via enzyme-linked immunosorbent assay. Cell counting kit-8 (CCK-8) assay and flow cytometry were employed to examine 16HBE cell viability and apoptosis, respectively. Western blotting was performed to examine the expression of TLR4, cleaved caspase-3, B-cell lymphoma-2 (Bcl-2), nuclear factor-kappa B (NF-κB) p65, Bcl-2-related X protein (Bax), phosphorylated (p)-NF-κB p65 and myeloid differentiation primitive-response protein 88 (MyD88) in 16HBE cells. Furthermore, the targeted relationship between TLR4 and miR-885-3p in 16HBE cells was determined through dual-luciferase reporter gene assay. Compared with healthy volunteers, miR-885-3p expression in acute asthma patients' plasma was significantly downregulated. In 16HBE cells, the stimulation of LPS reduced miR-885-3p expression. MiR-885-3p overexpression reduced LPS-stimulated 16HBE cell injury by enhancing cell viability, and suppressing the levels of inflammatory factors and apoptosis. Furthermore, TLR4 was identified as miR-885-3p's target gene. TLR4 overexpression weakened the impacts of miR-885-3p on LPS-stimulated cell injury and NF-κB-MyD88 signaling. In conclusion, miR-885-3p can reduce LPS-induced 16HBE cell damage, via targeting TLR4 to suppress the NF-κB-MyD88 pathway.

• miR-885-3p was up-regulated in response to TGEV-induced inflammation. • miR-885-3p inhibits TNF-α production via inhibiting NF-κB pathway. • Bcl-10 is the direct target gene of miR-885-3p. • TGEV-ORF3b upregulates miR-885-3p and downregulates Bcl-10 expression. Transmissible gastroenteritis (TGE) is an acute viral disease and characterized as severe acute inflammation response that leads to diarrhea, vomiting, and high lethality of piglets. Transmissible gastroenteritis virus (TGEV), a member of coronavirus, is the pathogen of TGE. We previously found NF-κB pathway was activated and 65 miRNAs were changed in response to inflammation caused by TGEV in cell line porcine intestinal epithelial cells-jejunum 2 (IPEC-J2). Bioinformatics results showed that these altered miRNAs were relevant to inflammation. In this study, the candidate targets of differentially expressed (DE) miRNAs were predicted and analyzed using Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Based on the results of KEGG analysis, miR-885-3p might participate in regulating activation of NF-κB pathway and TNF pathway. To study the function of miR-885-3p, miR-885-3p mimics and inhibitors were artificially synthesized and respectively used for overexpression and silence of miR-885-3p in cells. Our results showed that miR-885-3p inhibited NF-κB signaling pathway and tumor necrosis factor-α (TNF-α) production. B-cell CLL/lymphoma 10 (Bcl-10) was identified as the target of miR-885-3p, and promoted NF-κB pathway activation and TNF-α production. It was found that TGEV open reading frame 3b (TGEV-ORF3b) suppressed Bcl-10 expression, activation of NF-κB pathway, and TNF-α production by uniquely up-regulated miR-885-3p expression. Overall, the results indicated that TGEV-ORF3b counteracted NF-κB pathway and TNF-α via regulating miR-885-3p and Bcl-10. [ABSTRACT FROM AUTHOR]

Background: Type 1 diabetes (T1D) is a chronic autoimmune disease characterized by the progressive destruction of insulin‐production pancreatic β cells. Recently, microRNAs (miRNAs) have emerged as important regulators in T1D. The present study aimed to determine miR‐885‐3p expression in T1D patients and to examine the effects of miR‐885‐3p on the inflammatory response in human monocytes. Methods: Relevant gene expression levels were determined by a quantitative polymerase chain reaction; western blotting and enzyme‐linked immunosorbent assays determined the respective protein levels; and the interaction between miRNA and the downstream targets was evaluated using a luciferase reporter assay. Results: MiR‐885‐3p is down‐regulated and the levels of pro‐inflammatory cytokines are increased in peripheral blood mononuclear cells (PBMCs) from T1D patients compared to healthy controls. MiR‐885‐3p overexpression suppressed mRNA expression and secreted protein levels of pro‐inflammatory cytokines in THP‐1. A luciferase reporter assay showed that miR‐885‐3p directly targeted the 3'‐untranslated region of Toll‐like receptor 4 (TLR4) and miR‐885‐3p overexpression down‐regulated TLR4 expression in THP‐1 cells. The TLR4 mRNA expression level was increased in PBMCs isolated from T1D patients compared to heathy controls. TLR4 overexpression increased the secretion of pro‐inflammatory cytokines and enhanced the activity of NF‐κB signaling, and also attenuated the inhibitory effects of miR‐885‐3p overexpression on pro‐inflammatory cytokine secretion and the activity of NF‐κB signaling in THP‐1 cells. Conclusions: The present study identified the down‐regulation of miR‐885‐3p and up‐regulation of TLR4 in PBMCs isolated from T1D patients. Further mechanistic data demonstrated that miR‐885‐3p overexpression represses the production of pro‐inflammatory cytokines via targeting TLR4/NF‐κB signaling in THP‐1 cells. [ABSTRACT FROM AUTHOR]