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AbstractPurposeMethodsResultsConclusionThis study aimed to explore the expression profile of miR-185-5p in proliferative DR (PDR), and further evaluate its diagnostic value and possible mechanism of miR-185-5p in PDR.The level of miR-185-5p was detected by qRT-PCR. The ROC curve was established to estimate the diagnostic ability of miR-185-5p. Transwell experiment and cell counting kit-8 (CCK-8) assays were conducted to assess the effect of miR-185-5p on the migration and proliferation of human retinal endothelial cells (HRECs) induced by high glucose. Enzyme linked immunosorbent assay (ELISA) was used to detect the concentrations of inflammatory factors. The luciferase reporter gene experiment was used to prove the interaction between miR-185-5p and CXCR4.Compared to the control group, the expression of miR-185-5p was significantly up-regulated in both the type 2 diabetes mellitus (T2DM) group and the PDR groups, with higher levels in the PDR group than in the T2DM group. The ROC curve reveals that serum miR-185-5p can distinguish PDR patients from T2DM patients. MiR-185-5p levels in HRECs increased significantly after high glucose induction. High glucose induction also promoted the migration, proliferation and inflammation response of HRECs. However, when the intracellular miR-185-5p level was down-regulated by miR-185-5p inhibitor transfection, these effects were inhibited. The luciferase reporter gene assay showed that miR-185-5p directly targets CXCR4.The expression of miR-185-5p is out of balance in PDR and it may be involved in regulating the migration and proliferation of HRECs by regulating CXCR4. [ABSTRACT FROM AUTHOR]

AbstractThe aim of this study is to investigate the impact of sh-LncRNA ASB16-AS1 on doxorubicin (DOX) resistance in colorectal cancer (CRC). First, an in vitro study was conducted to investigate the effects of LncRNA ASB16-AS1, miR-185-5p, and TEAD1 on drug resistance in CRC cells. Subsequently, utilizing nanotechnology, poly(beta amino esters) (PBAE)/zeolitic imidazolate framework-8 (ZIF-8)@sh-LncRNA ASB16-AS1 nanoparticles (PZSNP) were synthesized and characterized, evaluating their cellular toxicity and hemolytic activity. Finally, a mouse subcutaneous tumor model was established by subcutaneous injection of SW480/DOX cell suspension to investigate the impact of PZSNP on the tumor. Under DOX treatment, downregulation of LncRNA ASB16-AS1, overexpression of miR-185-5p, or downregulation of TEAD1 suppressed the viability and proliferation of drug-resistant CRC cells while promoting apoptosis. Conversely, overexpression of LncRNA ASB16-AS1, inhibition of miR-185-5p, or overexpression of TEAD1 enhanced the viability and proliferation of drug-resistant CRC cells while inhibiting apoptosis. The synthesized PZSNP exhibited a spherical shape with an average particle size of 123.6 nm, possessed positive charge, displayed good stability. It effectively encapsulated shRNA and displayed low cellular toxicity and hemolytic activity. Under DOX treatment, significant tumor necrosis was observed in the PZSNP group, and tumor growth was suppressed without causing weight loss. LncRNA ASB16-AS1, miR-185-5p, and TEAD1 are involved in regulating cell viability, proliferation, and apoptosis, contributing to drug resistance in CRC cells. sh-LncRNA ASB16-AS1 enhances the sensitivity of CRC cells to DOX during treatment, and in vivo delivery of PZSNP may serve as an effective strategy to overcome chemotherapy resistance in CRC. [ABSTRACT FROM AUTHOR]

Na Yan,1 Xianggui Wang,1 Zufang Xu,1 Linling Zhong,1 Jiangyong Yang2 1Department of Vasculocardiology, Ganzhou People’s Hospital, Ganzhou, People’s Republic of China; 2Department of Cardiology, Ganzhou Hospital of Guangdong Provincial People’s Hospital, Ganzhou Municipal Hospital, Ganzhou, People’s Republic of ChinaCorrespondence: Jiangyong Yang, Department of Cardiology, Ganzhou Hospital of Guangdong Provincial People’s Hospital, Ganzhou Municipal Hospital, No. 49, Dagong Road, Ganzhou, 341000, People’s Republic of China, Tel +86-797-8208553, Email yangjy19861208@163.comIntroduction: Apigenin is a natural flavonoid compound with promising potential for the attenuation of myocardial hypertrophy (MH). The compound can also modulate the expression of miR-185-5p that both promote MH and suppress autophagy. The current attempts to explain the anti-MH effect of apigenin by focusing on changes in miR-185-5p-mediated autophagy.Methods: Hypertrophic symptoms were induced in rats using transverse aortic constriction (TAC) method and in cardiomyocytes using Ang II and then handled with apigenin. Changes in myocardial function and structure and cell viability and surface area were measured. The role of miR-185-5p in the anti-MH function of apigenin was explored by detecting changes in autophagic processes and miR-185-5p/SREBP2 axis.Results: TAC surgery induced weight increase, structure destruction, and collagen deposition in hearts of model rats. Ang II suppresses cardiomyocyte viability and increased cell surface area. All these impairments were attenuated by apigenin and were associated with the restored level of autophagy. At the molecular level, the expression of miR-185-5p was up-regulated by TAC, while the expression of SREBP2 was down-regulated, which was reserved by apigenin both in vivo and in vitro. The induction of miR-185-5p in cardiomyocytes could counteracted the protective effects of apigenin.Discussion: Collectively, the findings outlined in the current study highlighted that apigenin showed anti-MH effects. The effects were related to the inhibition of miR-185-5p and activation of SREBP, which contributed to the increased autophagy.Keywords: apigenin, autophagy, miR-185-5p, myocardial hypertrophy, SREBP2

Abstract Background The 22q11.2 deletion syndrome (22q11.2DS) is a microdeletion syndrome with highly variable phenotypic manifestations, even though most patients present the typical 3 Mb microdeletion, usually affecting the same ~ 106 genes. One of the genes affected by this deletion is DGCR8, which plays a crucial role in miRNA biogenesis. Therefore, the haploinsufficiency of DGCR8 due to this microdeletion can alter the modulation of the expression of several miRNAs involved in a range of biological processes. Results In this study, we used next-generation sequencing to evaluate the miRNAs profiles in the peripheral blood of 12 individuals with typical 22q11DS compared to 12 healthy matched controls. We used the DESeq2 package for differential gene expression analysis and the DIANA-miTED dataset to verify the expression of differentially expressed miRNAs in other tissues. We used miRWalk to predict the target genes of differentially expressed miRNAs. Here, we described two differentially expressed miRNAs in patients compared to controls: hsa-miR-1304-3p, located outside the 22q11.2 region, upregulated in patients, and hsa-miR-185-5p, located in the 22q11.2 region, which showed downregulation. Expression of miR-185-5p is observed in tissues frequently affected in patients with 22q11DS, and previous studies have reported its downregulation in individuals with 22q11DS. hsa-miR-1304-3p has low expression in blood and, thus, needs more validation, though using a sensitive technology allowed us to identify differences in expression between patients and controls. Conclusions Thus, lower expression of miR-185-5p can be related to the 22q11.2 deletion and DGCR8 haploinsufficiency, leading to phenotypic consequences in 22q11.2DS patients, while higher expression of hsa-miR-1304-3p might be related to individual genomic variances due to the heterogeneous background of the Brazilian population.

Background: The 22q11.2 deletion syndrome (22q11.2DS) is a microdeletion syndrome with highly variable phenotypic manifestations, even though most patients present the typical 3 Mb microdeletion, usually affecting the same ~ 106 genes. One of the genes affected by this deletion is DGCR8, which plays a crucial role in miRNA biogenesis. Therefore, the haploinsufficiency of DGCR8 due to this microdeletion can alter the modulation of the expression of several miRNAs involved in a range of biological processes. Results: In this study, we used next-generation sequencing to evaluate the miRNAs profiles in the peripheral blood of 12 individuals with typical 22q11DS compared to 12 healthy matched controls. We used the DESeq2 package for differential gene expression analysis and the DIANA-miTED dataset to verify the expression of differentially expressed miRNAs in other tissues. We used miRWalk to predict the target genes of differentially expressed miRNAs. Here, we described two differentially expressed miRNAs in patients compared to controls: hsa-miR-1304-3p, located outside the 22q11.2 region, upregulated in patients, and hsa-miR-185-5p, located in the 22q11.2 region, which showed downregulation. Expression of miR-185-5p is observed in tissues frequently affected in patients with 22q11DS, and previous studies have reported its downregulation in individuals with 22q11DS. hsa-miR-1304-3p has low expression in blood and, thus, needs more validation, though using a sensitive technology allowed us to identify differences in expression between patients and controls. Conclusions: Thus, lower expression of miR-185-5p can be related to the 22q11.2 deletion and DGCR8 haploinsufficiency, leading to phenotypic consequences in 22q11.2DS patients, while higher expression of hsa-miR-1304-3p might be related to individual genomic variances due to the heterogeneous background of the Brazilian population. [ABSTRACT FROM AUTHOR]

Istianah Purnamasari, Yuyun Sri Wahyuni, Hernawati Basir Department of Pharmacy, Faculty of Medicine and Health Science, Universitas Muhammadiyah Makassar, Makassar, South Sulawesi, IndonesiaCorrespondence: Istianah Purnamasari, Department of Pharmacy, Faculty of Medicine and Health Science, Universitas Muhammadiyah Makassar, Makassar, South Sulawesi, Indonesia, Email istianahpurnamasari@unismuh.ac.id

OBJECTIVE: Late Fetal Growth Restriction (LFGR), a condition associated with perinatal and long-term neurodevelopmental problems, is caused by inadequate placental transfer of oxygen and nutrients to the fetus. This preliminary study evaluates the expression profiles and clinical significance of hypoxia-sensitive microRNA-210 (miR-210) and microRNA-185-5p (miR-185-5p) in placental and maternal circulation. The potential clinical implications of this research could significantly enhance the management of LFGR. STUDY DESIGN: In this prospective cohort study conducted at the Gynecology and Obstetrics Clinic of Trakya University Faculty of Medicine Hospital, miR-210 and miR-185-5p expression was evaluated in the placenta of healthy (n=30) and LFGR (n=30) cases. In healthy (n=15) and LFGR (n=15) cases, miRNA levels in maternal plasma were studied. We determined these two miRNAs using Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR) and statistically analyzed them with clinical data to determine robustness. We provided a comprehensive evaluation. RESULTS: Both the placenta and maternal plasma samples exhibited significant correlations with miR- 210 and miR-185-5p expression (r=0.615; r=0.771, p<0.01). The expression levels in both placenta and plasma were significantly higher in the LFGR group. Placental miR-210 demonstrated an AUC (0.908), sensitivity (93.3%), and specificity (96.7%), while plasma miR-210 showed AUC (0.738), sensitivity (86.7%), and specificity (66.7%). In contrast, placental miRNA 185-5p displayed a diagnostic performance with AUC (0.926), sensitivity (76.1%), and specificity (63.8%); plasma miRNA 185-5p exhibited an AUC (0.778), sensitivity (86.7%), and specificity (86.7%). However, placental miR-210 and miR-185-5p did not fully correlate with birth weight, placental weight, and fetal cerebroplacental Doppler ratio. CONCLUSION: Placental miRNA-210 showed superior diagnostic performance over placental miRNA 185-5p in LFGR pregnancies. However, it is crucial to emphasize that further studies are needed to validate these findings and correlate them with clinical data, underscoring the ongoing importance of research in this field. [ABSTRACT FROM AUTHOR]

Introduction: Self‐repair of spinal cord injury (SCI) has been found in humans and experimental animals with partial recovery of neurological functions. However, the regulatory mechanisms underlying the spontaneous locomotion recovery after SCI are elusive. Aims: This study was aimed at evaluating the pathological changes in injured spinal cord and exploring the possible mechanism related to the spontaneous recovery. Results: Immunofluorescence staining was performed to detect GAP43 expression in lesion site after spinal cord transection (SCT) in rats. Then RNA sequencing and gene ontology (GO) analysis were employed to predict lncRNA that correlates with GAP43. LncRNA smart‐silencing was applied to verify the function of lncRNA vof16 in vitro, and knockout rats were used to evaluate its role in neurobehavioral functions after SCT. MicroRNA sequencing, target scan, and RNA22 prediction were performed to further explore the underlying regulatory mechanisms, and miR‐185‐5p stands out. A miR‐185‐5p site‐regulated relationship with GAP43 and vof16 was determined by luciferase activity analysis. GAP43‐silencing, miR‐185‐5p‐mimic/inhibitor, and miR‐185‐5p knockout rats were also applied to elucidate their effects on spinal cord neurite growth and neurobehavioral function after SCT. We found that a time‐dependent increase of GAP43 corresponded with the limited neurological recovery in rats with SCT. CRNA chip and GO analysis revealed lncRNA vof16 was the most functional in targeting GAP43 in SCT rats. Additionally, silencing vof16 suppressed neurite growth and attenuated the motor dysfunction in SCT rats. Luciferase reporter assay showed that miR‐185‐5p competitively bound the same regulatory region of vof16 and GAP43. Conclusions: Our data indicated miR‐185‐5p could be a detrimental factor in SCT, and vof16 may function as a ceRNA by competitively binding miR‐185‐5p to modulate GAP43 in the process of self‐recovery after SCT. Our study revealed a novel vof16‐miR‐185‐5p‐GAP43 regulatory network in neurological self‐repair after SCT and may underlie the potential treatment target for SCI. [ABSTRACT FROM AUTHOR]

Background: For IgA nephropathy (IgAN), tubular atrophy/interstitial fibrosis is the most important prognostic pathological indicator in the mesangial and endocapillary hypercellularity, segmental sclerosis, interstitial fibrosis/tubular atrophy, and presence of crescents (MEST-C) score. The identification of noninvasive biomarkers for tubular atrophy/interstitial fibrosis would aid clinical monitoring of IgAN progression and improve patient prognosis. Methods: The study included 188 patients with primary IgAN in separate confirmation and validation cohorts. The associations of miR-92a-3p, miR- 425-5p, and miR-185-5p with renal histopathological lesions and prognosis were explored using Spearman correlation analysis and Kaplan-Meier survival curves. Bioinformatics analysis and dual luciferase experiments were used to identify hub genes for miR-185-5p. The fibrotic phenotypes of tubular epithelial cells were evaluated in vivo and in HK-2 cells. Results: miRNA sequencing and cohort validation revealed that the expression levels of miR-92a-3p, miR-425-5p, and miR-185-5p in urine were significantly increased among patients with IgAN; these levels could predict the extent of tubular atrophy/interstitial fibrosis in such patients. The combination of the three biomarkers resulted in an area under the receiver operating characteristic curve of 0.742. The renal prognosis was significantly worse in the miR-185-5p high expression group than in the low expression group (P=0.003). Renal tissue in situ hybridization, bioinformatics analysis, and dual luciferase experiments confirmed that miR-185-5p affects prognosis in patients with IgAN mainly by influencing expression of the target gene tight junction protein 1 (TJP1) in renal tubular epithelial cells. In vitro experiment revealed that an miR-185-5p mimic could reduce TJP1 expression in HK-2 cells, while increasing the levels of a-smooth muscle actin, fibronectin, collagen I, and collagen III; these changes promoted the transformation of renal tubular epithelial cells to a fibrotic phenotype. An miR-185-5p inhibitor can reverse the fibrotic phenotype in renal tubular epithelial cells. In a unilateral ureteral obstruction model, the inhibition of miR-185-5p expression alleviated tubular atrophy/interstitial fibrosis. Conclusion: Urinary miR-185-5p, a non-invasive biomarker of tubular atrophy/ interstitial fibrosis in IgAN, may promote the transformation of renal tubular epithelial cells to a fibrotic phenotype via TJP1. [ABSTRACT FROM AUTHOR]

BACKGROUND: The pathogenesis of periodontitis is still unclear, and its therapeutic target remains to be explored. Recent studies have showed that miR-185- 5p has anti-inflammatory effects in many diseases. However, it is unclear whether miR-185-5p can influence the inflammatory response in periodontitis. OBJECTIVE: To investigate the effect of regulating miR-185-5p expression on inflammatory response in experimental periodontitis in mice. METHODS: Twenty-four C57B/6J mice were randomly divided into normal control group, periodontitis control group, periodontitis miR-NC group and periodontitis miR-mimics group (n=6 per group). Except for the normal control group, other groups were induced into experimental periodontitis. Four weeks after modeling, in the periodontitis miR-mimics group, 100 μL of normal saline containing 0.5 mmol/L miR-185-5p mimics was injected into the proximal and distal gingival papillae of bilateral mandibular second molars of mice. In the periodontitis miR-NC group, 100 μL of normal saline containing 0.5 mmol/L negative control miR-NC was injected. In the normal control group and the periodontitis control group, the same amount of normal saline was injected. After 8 weeks of administration, all mice were sacrificed. Micro-CT was used to detect the left mandibles, and qRT-PCR was used to detect the gene expression of miR-185-5p, marginal zone B and B-1 cell-specific protein (MZB1), tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) in the gingiva of left mandibles. Immunohistochemical staining was used to detect the protein expression of MZB1, phospho-p65 NF-κB (pp65), TNF-α and IL-6 in the gingiva of right mandibles. RESULTS AND CONCLUSION: Micro-CT examination results showed that compared with the periodontitis control group and periodontitis miR-NC group, alveolar bone loss in the periodontitis miR-mimics group was decreased (P < 0.05). qRT-PCR results showed that the expression of miR-185-5p in the periodontitis miR-mimics group was higher than that in the normal control group, periodontitis control group, and periodontitis miR-NC group (P < 0.05). Compared with the periodontitis control group and periodontitis miR-NC group, the mRNA expression of MZB1, TNF-α, and IL-6 was decreased in the periodontitis miR-mimics group (P < 0.05). Immunohistochemical staining results showed that compared with the periodontitis control group and periodontitis miR-NC group, the protein expression of MZB1, pp65, TNF-α and IL-6 was decreased in the periodontitis miR-mimics group (P < 0.05). Therefore, increasing the expression of miR-185-5p can inhibit the inflammatory response in periodontitis and the resorption of alveolar bone, which may be associated with the suppression of MZB1 and nuclear fctor-κB pathway in the gingival tissue. [ABSTRACT FROM AUTHOR]

The healing of skin wounds is a highly coordinated multi-step process that occurs after trauma including surgical incisions, thermal burns, and chronic ulcers. In this study, the authors investigated lncRNA FOXD2-AS1 function in adipose mesenchymal exosomes from ADMSCs that were successfully extracted. Highly expressed lncRNA FOXD2-AS1 in ADMSCs-exosomes accelerated HaCaT cell migration and proliferation. LncRNA FOXD2-AS1 negatively targeted miR-185-5p, and miR-185-5p negatively targeted ROCK2. Highly expressed lncRNA FOXD2-AS1 in ADMSCs-exosomes promoted HaCaT cell migration and proliferation via down-regulating miR-185-5p and further up-regulating ROCK2. In conclusion, LncRNA FOXD2-AS1 overexpression in ADMSCs derived exosomes might accelerate HaCaT cell migration and proliferation via modulating the miR-185-5p/ROCK2 axis. [ABSTRACT FROM AUTHOR]

Abstract Background Lung cancer (LC) is one of the most frequent cancers worldwide, as well as the leading cause of cancer-related death. Non-small cell lung cancer (NSCLC, which accounts for 85% of occurrences) is the main type of LC. MiRNAs appear to play a role in the occurrence and progression of many malignancies, according to mounting data. The underlying mechanism of miRNAs in regulating NSCLC cell biological activity and progression, on the other hand, is still being investigated. Methods QRT-PCR were used to detect miR-185-5p expression and YWHAZ mRNA in NSCLC. The CCK-8 assay was used to determine the tumor cells’ ability to proliferate. Transwall assay was used to test the migratory and invasive properties of cells. Cell apoptosis was detected using flow cytometry. Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ), E-Cadherin, N-Cadherin and cleaved-caspase3 protein expression were assessed using Western Blot. The bioinformatics analysis software StarBase2.0 predicted miR-185-5p downstream targets. To confirm the target association between miR-185-5p and YWHAZ, a luciferase experiment was used. In addition, an NCl-H1299 xenograft model was created to assess the anti-tumor impact of miR-185-5p in vivo. The expression level of YWHAZ in tumor tissues of small xenograft tumor model was detected by immunohistochemistry assay. Results Decreased miR-185-5p expression levels were observed in NSCLC. In vitro, over-expressed miR-185-5p decreased cell viability, proliferation, invasion/migration, and induced cell apoptosis, while inhibiting tumor growth in vivo. Dual-luciferase gene experiments confirmed that YWHAZ binds to miR-185-5p. Overexpression of YWHAZ partially restored the inhibitory effects of miR-185-5p on cell behaviors. Conclusion MiR-185-5p was down-regulated in NSCLC, and that overexpressed miR-185-5p inhibited malignant behaviors of cells and tumor growth by negatively regulating YWHAZ.

Objective·To explore the regulatory effect of miR-185-5p on autophagy in Mycobacterium-infected macrophages and its influence on the survival of intracellular Mycobacterium.Methods·Macrophages derived from human monocytic-leukemia (THP-1) cells were infected with Bacillus Calmette-Guérin (BCG) strain. Exosomes from cell culture supernatant were isolated by ultracentrifugation, and exosome surface marker proteins, size and morphology were detected by Western blotting, nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM), respectively. The expression level of miR-185-5p in both macrophages and exosomes was detected by real time quantitative polymerase chain reaction (qPCR). The exosomes were labeled with the fluorescent dye PKH26 and co-cultured with macrophages to observe the phagocytosis of exosomes by macrophages. Subsequently, the overexpression and inhibition of miR-185-5p in macrophages were achieved by transfecting cells with miR-185-5p mimic and inhibitor, and the influence of miR-185-5p on intracellular BCG survival was detected by colony-forming units (CFU) assay. Autophagy marker microtubule-associated protein 1 light chain 3 (LC3) protein was detected by Western blotting.The effects of overexpression and inhibition of miR-185-5p and the use of autophagy inhibitor chloroquine (CQ) on macrophage autophagy were studied. The SensGFP-StubRFP-LC3 lentvirus was further employed to detect the effect of miR-185-5p on autophagic flux. Finally, TargetScan, miRDB and Starbase databases were used to predict the target genes of miR-185-5p, and the Database for Annotation, Visualization and Integrated Discovery (DAVID) was used for Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis of these predicted target genes.Results·The macrophage exosomes were isolated by ultracentrifugation. Exosome marker proteins cluster differentiation 9 (CD9) and tumor susceptibility gene 101 (TSG101) were successfully detected by Western blotting. The exosomes had a bilayer membrane with a diameter of approximately 150 nm through TEM and NTA identification. After BCG infection, the expression of miR-185-5p was up-regulated in both macrophages and exosomes (P=0.000), and showed a time-dependent increase in macrophages. Exosomes, which were co-cultured with macrophages, can be phagocytized by macrophages. Compared with respective control of mimic and inhibitor, mimic significantly up-regulated miR-185-5p expression (P=0.000), while inhibitor inhibited miR-185-5p expression (P=0.002). miR-185-5p mimic significantly increased the number of CFU of intracellular BCG (P=0.000), while miR-185-5p inhibitor decreased the number of CFU (P=0.041). BCG infection up-regulated the expression of autophagy protein LC3Ⅱ in macrophages, while LC3Ⅱ expression was decreased by miR-185-5p mimic and increased by miR-185-5p inhibitor. CQ treatment combined with miR-185-5p inhibitor transfection still significantly increased LC3Ⅱ expression in macrophages. The SensGFP-StubRFP-LC3 lentivirus was further used to detect autophagy flux, and it was suggested that, compared with that in the respective control group, mimic significantly reduced the formation of autophagosomes (P=0.034), while inhibitor increased the formation of autophagosomes (P=0.042). Through target genes prediction and GO and KEGG functional enrichment analysis, it was found that miR-185-5p might play a role in autophagy regulation by targeting nuclear receptor subfamily 1 group D member 1 (NR1D1), Ras-related protein Rab-35 (RAB35), cell division control protein 42 homolog (CDC42), etc.Conclusion·Mycobacterium infection can induce the up-regulation of miR-185-5p in both macrophages and exosomes, and miR-185-5p can inhibit the autophagy process of macrophages by inhibiting the formation of autophagosomes, which can promote the growth and survival of intracellular Mycobacterium.

Abstract Background Circ‐ZKSCAN1 has been found to accelerate non‐small cell lung cancer (NSCLC) progression; however, the role and mechanism of circ‐ZKSCAN1 in lung adenocarcinoma (LUAD) cisplatin (DDP) resistance remain unclear. Methods Levels of genes and proteins were examined using qRT‐PCR. Functional experiments were performed using CCK‐8 assay, flow cytometry, transwell assay and xenograft model assay, respectively. Glucose metabolism was calculated by detecting glucose consumption, lactate production, ATP and HK‐2 levels. The interaction between miR‐185‐5p and circ‐ZKSCAN1 or transgelin 2 (TAGLN2) was validated by dual‐luciferase reporter assay. Results Circ‐ZKSCAN1 was highly expressed in DDP‐resistant LUAD tissues and cell lines, and circ‐ZKSCAN1 knockdown weakened DDP resistance and suppressed cell viability, migration, invasion, and glycolysis in LUAD. Circ‐ZKSCAN1 acted as a sponge of miR‐185‐5p, and the regulatory effects of circ‐ZKSCAN1 knockdown on LUAD were reversed by miR‐185‐5p downregulation. Meanwhile, miR‐185‐5p directly targeted TAGLN2, and performed anticancer effects by regulating TAGLN2. Importantly, silencing of circ‐ZKSCAN1 hindered tumor growth and promoted DDP sensitivity in vivo via regulating miR‐185‐5p and TAGLN2. Conclusion Circ‐ZKSCAN1 promoted LUAD tumorigenesis and DDP resistance by regulating miR‐185‐5p/TAGLN2 axis.

ABSTRACTThe healing of skin wounds is a highly coordinated multi-step process that occurs after trauma including surgical incisions, thermal burns, and chronic ulcers. In this study, the authors investigated lncRNA FOXD2-AS1 function in adipose mesenchymal exosomes from ADMSCs that were successfully extracted. Highly expressed lncRNA FOXD2-AS1 in ADMSCs-exosomes accelerated HaCaT cell migration and proliferation. LncRNA FOXD2-AS1 negatively targeted miR-185-5p, and miR-185-5p negatively targeted ROCK2. Highly expressed lncRNA FOXD2-AS1 in ADMSCs-exosomes promoted HaCaT cell migration and proliferation via down-regulating miR-185-5p and further up-regulating ROCK2. In conclusion, LncRNA FOXD2-AS1 overexpression in ADMSCs derived exosomes might accelerate HaCaT cell migration and proliferation via modulating the miR-185-5p/ROCK2 axis.

Background: This study aims to explore whether serum miR-185-5p levels are related to the injury severity and prognosis of traumatic brain injury patients. Methodi: Serum miR-185-5p level was quantified in 120 TBI patients. The Glasgow Coma Scale (GCS) was used to grade the damage, and the Glasgow Outcome Scale (GOS) was used to evaluate the prognosis 3 months after TBI. Pearson correlation analysis was performed to determine the relationship between serum miR-185-5p level and injury severity and prognosis, and the value of serum miR-185-5p level to assess injury severity and prognosis was evaluated by receiver operating characteristic (ROC) curve. Results: Serum miR-185-5p level in moderate and severe TBI patients was higher than in mild TBI patients, and serum miR-185-5p was closely related to GCS score and GOS score. Serum miR-185-5p level higher than 0.36 could distinguish patients with mild to moderate TBI injury, with 72.97% sensitivity and 97.62% specificity, while that higher than 0.43 had 46.34% sensitivity and 91.89% specificity to distinguish moderate to severe TBI patients. Moreover, serum miR-185-5p levels higher than 0.36, with a sensitivity of 96.30% and a specificity of 60.24%, distinguished the poor prognosis of TBI patients. Serum miR- 185-5p level was an independent predictor of poor prognosis in TBI patients after 3 months and was effective in discriminating adverse outcomes at 3 months. Conclusions: Serum miR-185-5p level was significantly correlated with 3-month injury and adverse prognosis in TBI patients, suggesting that serum miR-185-5p level may be a biomarker that provides supplementary prognostic information and can be used to identify the risk of adverse prognosis in TBI patients. [ABSTRACT FROM AUTHOR]

To explore the underlying mechanism of lncRNA MALAT1 in the pathogenesis of diabetic cardiomyopathy (DCM). DCM models were confirmed in db/db mice. MiRNAs in myocardium were detected by miRNA sequencing. The interactions of miR‐185‐5p with MALAT1 and RhoA were validated by dual‐luciferase reporter assays. Primary neonatal cardiomyocytes were cultured with 5.5 or 30 mmol/L D‐glucose (HG) in the presence or absence of MALAT1‐shRNA and fasudil, a ROCK inhibitor. MALAT1 and miR‐185‐5p expression were determined by real‐time quantitative PCR. The apoptotic cardiomyocytes were evaluated using flow cytometry and TUNEL staining. SOD activity and MDA contents were measured. The ROCK activity, phosphorylation of Drp1S616, mitofusin 2 and apoptosis‐related proteins were analysed by Western blotting. Mitochondrial membrane potential was examined by JC‐1. MALAT1 was significantly up‐regulated while miR‐185‐5p was down‐regulated in myocardium of db/db mice and HG‐induced cardiomyocytes. MALAT1 regulated RhoA/ROCK pathway via sponging miR‐185‐5p in cardiomyocytes in HG. Knockdown of MALAT1 and fasudil all inhibited HG‐induced oxidative stress, and alleviated imbalance of mitochondrial dynamics and mitochondrial dysfunction, accompanied by reduced cardiomyocyte apoptosis. MALAT1 activated the RhoA/ROCK pathway via sponging miR‐185‐5p and mediated HG‐induced oxidative stress, mitochondrial damage and apoptosis of cardiomyocytes in mice. [ABSTRACT FROM AUTHOR]

Chemoresistance to paclitaxel (PTX) is one of the main reasons for treatment failure and poor prognosis in patients with advanced ovarian cancer. Therefore, it is imperative to explore the mechanisms related to chemotherapy resistance in ovarian cancer to find potential therapeutic targets. Circular RNAs (circRNAs) play important roles in cancer development and progression. However, their biological functions and clinical significance in ovarian cancer have not been fully elucidated. Therefore, in this study, we aimed to investigate the function and underlying mechanism of hsa_circ_0002782 (circSLC39A8), identified by circRNA sequencing, in regulating PTX resistance. The effects of circSLC39A8 on PTX resistance was assessed by cell viability, colony formation, flow cytometry assays and an in vivo subcutaneous xenografted tumor mouse model. RNA immunoprecipitation and dual-luciferase reporter assays were performed to verify the interaction between circSLC39A8 and the miR-185–5p/BMF signal axis. We found that circSLC39A8 was downregulated in PTX-resistant ovarian cancer cells and tissues, and its low expression was associated with poor prognosis. Biologically, circSLC39A8 knockdown promoted PTX resistance in vitro and in vivo, while circSLC39A8 overexpression showed the opposite effect. Mechanistically, circSLC39A8, acting as an endogenous sponge for miR-185–5p, could relieve the inhibition of miR-185–5p on the expression of its downstream target, BMF; thus enhancing the sensitivity of ovarian cancer to PTX. Our findings demonstrate that circSLC39A8 can promote PTX sensitivity by regulating the miR-185–5p/BMF axis. This may be a valuable prognostic biomarker and a promising therapeutic target for patients with ovarian cancer.

Background: Lung cancer (LC) is one of the most frequent cancers worldwide, as well as the leading cause of cancer-related death. Non-small cell lung cancer (NSCLC, which accounts for 85% of occurrences) is the main type of LC. MiRNAs appear to play a role in the occurrence and progression of many malignancies, according to mounting data. The underlying mechanism of miRNAs in regulating NSCLC cell biological activity and progression, on the other hand, is still being investigated. Methods: QRT-PCR were used to detect miR-185-5p expression and YWHAZ mRNA in NSCLC. The CCK-8 assay was used to determine the tumor cells' ability to proliferate. Transwall assay was used to test the migratory and invasive properties of cells. Cell apoptosis was detected using flow cytometry. Tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ), E-Cadherin, N-Cadherin and cleaved-caspase3 protein expression were assessed using Western Blot. The bioinformatics analysis software StarBase2.0 predicted miR-185-5p downstream targets. To confirm the target association between miR-185-5p and YWHAZ, a luciferase experiment was used. In addition, an NCl-H1299 xenograft model was created to assess the anti-tumor impact of miR-185-5p in vivo. The expression level of YWHAZ in tumor tissues of small xenograft tumor model was detected by immunohistochemistry assay. Results: Decreased miR-185-5p expression levels were observed in NSCLC. In vitro, over-expressed miR-185-5p decreased cell viability, proliferation, invasion/migration, and induced cell apoptosis, while inhibiting tumor growth in vivo. Dual-luciferase gene experiments confirmed that YWHAZ binds to miR-185-5p. Overexpression of YWHAZ partially restored the inhibitory effects of miR-185-5p on cell behaviors. Conclusion: MiR-185-5p was down-regulated in NSCLC, and that overexpressed miR-185-5p inhibited malignant behaviors of cells and tumor growth by negatively regulating YWHAZ. [ABSTRACT FROM AUTHOR]

Circular RNA (circRNA) is key regulator of diabetic nephropathy (DN) progression. However, the role of circ_0008529 in DN progression remains to be better deciphered. Cell viability, cell cycle, apoptosis and inflammation were measured by MTS assay, flow cytometry and corresponding assay kits. RT-qPCR was used to assess the expression of circ_0008529, miR-185-5p and SMAD family member 2 (SMAD2). Also, western blotting was performed to measure protein expression. Target relationship was validated by RNA pull-down assay, dual-luciferase reporter assay and RNA immunoprecipitation assay. Urinary exosome was isolated using ultracentrifugation method and identified by transmission electron microscopy. Receiver operating characteristic curve was used to analyze the diagnostic value of circ_0008529 in DN patients. Circ_0008529 and SMAD2 were upregulated, while miR-185-5p was downregulated in high glucose (HG)-induced renal tubular HK-2 cells. Under HG treatment, cell viability and cell cycle process were suppressed, while apoptosis, inflammation and extracellular matrix accumulation were enhanced. However, interfering circ_0008529 could attenuate HG-induced effects, and this protection was abated by miR-185 inhibition or SMAD2 re-expression. Mechanically, circ_0008529 and SMAD2 were competing endogenous RNAs for miR-185-5p via target binding, and circ_0008529 regulated SMAD2 expression via miR-185-5p. Notably, circ_0008529 expression was upregulated in urinary exosomes of DN patients, and showed diagnostic value (Sensitivity: 70.21%; Specificity: 86.67%). Circ_0008529 might be a potential target for DN, which regulated DN progression via miR-185-5p/SMAD2 pathway. [ABSTRACT FROM AUTHOR]

Background: Circ‐ZKSCAN1 has been found to accelerate non‐small cell lung cancer (NSCLC) progression; however, the role and mechanism of circ‐ZKSCAN1 in lung adenocarcinoma (LUAD) cisplatin (DDP) resistance remain unclear. Methods: Levels of genes and proteins were examined using qRT‐PCR. Functional experiments were performed using CCK‐8 assay, flow cytometry, transwell assay and xenograft model assay, respectively. Glucose metabolism was calculated by detecting glucose consumption, lactate production, ATP and HK‐2 levels. The interaction between miR‐185‐5p and circ‐ZKSCAN1 or transgelin 2 (TAGLN2) was validated by dual‐luciferase reporter assay. Results: Circ‐ZKSCAN1 was highly expressed in DDP‐resistant LUAD tissues and cell lines, and circ‐ZKSCAN1 knockdown weakened DDP resistance and suppressed cell viability, migration, invasion, and glycolysis in LUAD. Circ‐ZKSCAN1 acted as a sponge of miR‐185‐5p, and the regulatory effects of circ‐ZKSCAN1 knockdown on LUAD were reversed by miR‐185‐5p downregulation. Meanwhile, miR‐185‐5p directly targeted TAGLN2, and performed anticancer effects by regulating TAGLN2. Importantly, silencing of circ‐ZKSCAN1 hindered tumor growth and promoted DDP sensitivity in vivo via regulating miR‐185‐5p and TAGLN2. Conclusion: Circ‐ZKSCAN1 promoted LUAD tumorigenesis and DDP resistance by regulating miR‐185‐5p/TAGLN2 axis. [ABSTRACT FROM AUTHOR]

Objective ·To explore the regulatory effect of miR-185-5p on autophagy in Mycobacterium-infected macrophages and its influence on the survival of intracellular Mycobacterium. Methods ·Macrophages derived from human monocytic-leukemia (THP-1) cells were infected with Bacillus Calmette-Guérin (BCG) strain. Exosomes from cell culture supernatant were isolated by ultracentrifugation, and exosome surface marker proteins, size and morphology were detected by Western blotting, nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM), respectively. The expression level of miR-185-5p in both macrophages and exosomes was detected by real time quantitative polymerase chain reaction (qPCR). The exosomes were labeled with the fluorescent dye PKH26 and co-cultured with macrophages to observe the phagocytosis of exosomes by macrophages. Subsequently, the overexpression and inhibition of miR-185-5p in macrophages were achieved by transfecting cells with miR-185-5p mimic and inhibitor, and the influence of miR-185-5p on intracellular BCG survival was detected by colony-forming units (CFU) assay. Autophagy marker microtubule-associated protein 1 light chain 3 (LC3) protein was detected by Western blotting. The effects of overexpression and inhibition of miR-185-5p and the use of autophagy inhibitor chloroquine (CQ) on macrophage autophagy were studied. The SensGFP-StubRFP-LC3 lentvirus was further employed to detect the effect of miR-185-5p on autophagic flux. Finally, TargetScan, miRDB and Starbase databases were used to predict the target genes of miR-185-5p, and the Database for Annotation, Visualization and Integrated Discovery (DAVID) was used for Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis of these predicted target genes. Results ·The macrophage exosomes were isolated by ultracentrifugation. Exosome marker proteins cluster differentiation 9 (CD9) and tumor susceptibility gene 101 (TSG101) were successfully detected by Western blotting. The exosomes had a bilayer membrane with a diameter of approximately 150 nm through TEM and NTA identification. After BCG infection, the expression of miR-185-5p was up-regulated in both macrophages and exosomes (P=0.000), and showed a time-dependent increase in macrophages. Exosomes, which were co-cultured with macrophages, can be phagocytized by macrophages. Compared with respective control of mimic and inhibitor, mimic significantly up-regulated miR-185-5p expression (P=0.000), while inhibitor inhibited miR-185-5p expression (P=0.002). miR-185-5p mimic significantly increased the number of CFU of intracellular BCG (P=0.000), while miR-185-5p inhibitor decreased the number of CFU (P=0.041). BCG infection up-regulated the expression of autophagy protein LC3Ⅱ in macrophages, while LC3Ⅱ expression was decreased by miR-185-5p mimic and increased by miR-185-5p inhibitor. CQ treatment combined with miR-185-5p inhibitor transfection still significantly increased LC3Ⅱ expression in macrophages. The SensGFP-StubRFP-LC3 lentivirus was further used to detect autophagy flux, and it was suggested that, compared with that in the respective control group, mimic significantly reduced the formation of autophagosomes (P=0.034), while inhibitor increased the formation of autophagosomes (P=0.042). Through target genes prediction and GO and KEGG functional enrichment analysis, it was found that miR-185-5p might play a role in autophagy regulation by targeting nuclear receptor subfamily 1 group D member 1 (NR1D1), Ras-related protein Rab-35 (RAB35), cell division control protein 42 homolog (CDC42), etc. Conclusion ·Mycobacterium infection can induce the up-regulation of miR-185-5p in both macrophages and exosomes, and miR-185-5p can inhibit the autophagy process of macrophages by inhibiting the formation of autophagosomes, which can promote the growth and survival of intracellular Mycobacterium. [ABSTRACT FROM AUTHOR]

Background: This study aims to explore whether serum miR-185-5p levels are related to the injury severity and prognosis of traumatic brain injury patients. Methods: Serum miR-185-5p level was quantified in 120 TBI patients. The Glasgow Coma Scale (GCS) was used to grade the damage, and the Glasgow Outcome Scale (GOS) was used to evaluate the prognosis 3 months after TBI. Pearson correlation analysis was performed to determine the relationship between serum miR-185-5p level and injury severity and prognosis, and the value of serum miR-185-5p level to assess injury severity and prognosis was evaluated by receiver operating characteristic (ROC) curve. Results: Serum miR-185-5p level in moderate and severe TBI patients was higher than in mild TBI patients, and serum miR-185-5p was closely related to GCS score and GOS score. Serum miR-185-5p level higher than 0.36 could distinguish patients with mild to moderate TBI injury, with 72.97% sensitivity and 97.62% specificity, while that higher than 0.43 had 46.34% sensitivity and 91.89% specificity to distinguish moderate to severe TBI patients. Moreover, serum miR-185-5p levels higher than 0.36, with a sensitivity of 96.30% and a specificity of 60.24%, distinguished the poor prognosis of TBI patients. Serum miR185-5p level was an independent predictor of poor prognosis in TBI patients after 3 months and was effective in discriminating adverse outcomes at 3 months. Conclusions: Serum miR-185-5p level was significantly correlated with 3-month injury and adverse prognosis in TBI patients, suggesting that serum miR-185-5p level may be a biomarker that provides supplementary prognostic information and can be used to identify the risk of adverse prognosis in TBI patients.

Circular RNA (circRNA) has been confirmed to be the key regulators of diabetic nephropathy (DN) progression. However, the role of circ_0114428 in the DN progression remains unclear. Glomerular mesangial cells (GMCs) were treated with high glucose (HG) to mimic DN cell models in vitro. The expression levels of circ_0114428, microRNA (miR)-185-5p, and SMAD3 mRNA were examined by quantitative real-time PCR. Cell proliferation ability was detected by MTT assay, EdU staining and flow cytometry. The protein levels of proliferation marker, fibrosis markers, epithelial-mesenchymal transition (EMT) markers and SMAD3 were measured by western blot assay. The interaction between miR-185-5p and circ_0114428 or SMAD3 was confirmed via dual-luciferase reporter assay, RIP assay and RNA pull-down assay. Our data showed that circ_0114428 was upregulated in HG-induced GMCs. Circ_0114428 overexpression could aggravate the promotion effect of HG on the proliferation, fibrosis and EMT process of GMCs, while its knockdown had an opposite effect. In the terms of mechanisms, circ_0114428 could sponge miR-185-5p to regulate SMAD3. MiR-185-5p inhibitor could reverse the suppressive effect of circ_0114428 knockdown on the proliferation, fibrosis and EMT process in HG-induced GMCs. Also, SMAD3 overexpression abolished the inhibition of miR-185-5p on the proliferation, fibrosis and EMT process in HG-induced GMCs. Taken together, our data suggested that circ_0114428 might promote DN progression by regulating the miR-185-5p/SMAD3 axis.

Objective: Intestinal ischemia-reperfusion (II/R) is a common pathological injury in clinic, and the systemic inflammatory response it causes will lead to multiple organ damage and functional failure. miR-185-5p has been reported to be a regulator of inflammatory response and autophagy, but whether it participates in the regulation of autophagy in II/R is still unclear. Therefore, we aimed to explore the mechanism of miR-185-5p regulating intestinal barrier injury in (II/R). Methods: Caco-2 cells was induced by oxygen-glucose deprivation/reoxygenation (OGD/R) to establish II/R model. The superior mesenteric artery of C57BL/6 mice was clamped for 45 min and then subjected to reperfusion for 4 h for the establishment of II/R mice model. miR-185-5p mimic, miR-185-5p inhibitor, pcDNA-autophagy-related 101 (ATG101) were respectively transfected into Caco-2 cells. Real-time quantitative polymerase chain reaction (RT-qPCR) was performed to assess miR-185-5p expression. Western blot detected the level of ATG101 and tight junction-associated proteins ZO1, Occludin, E-cadherin, β-catenin, as well as autophagy markers ATG5, ATG12, LC3Ⅰ/Ⅱ, Beclin1 and SQSTM1. Transepithelial electrical resistance (TEER) values was detected by a resistance meter. FITC-Dextran was performed to measure cell permeability. 5-ethynyl-2’-deoxyuridine (EDU) staining measured cell proliferation. Transmission electron microscope was conducted to observe autophagosomes. Hematoxylin & eosin (H&E) staining observed the damage of mice intestinal. Immunohistochemistry (IHC) measured the percentage of ki67 positive cells. TdT-mediated dUTP nick-end labeling (TUNEL) assay assessed cell apoptosis in intestinal tissues of II/R. Dual-luciferase assay verified the targeting relationship between miR-185-5p and ATG101.Results miR-185-5p was overexpressed in OGD/R-induced Caco-2 cells and intestinal tissues of II/R mice. Knocking down miR-185-5p markedly promoted autophagy and TEER values, reduced cell permeability, and alleviated intestinal barrier damage. ATG101 was a target of miR-185-5p, and overexpression of ATG101 promoted autophagy and dampened OGD/R-induced intestinal barrier damage. Overexpression of miR-185-5p reversed the effect of overexpressed ATG101 on OGD/R-induced Caco-2 cells. Conclusion: Knockdown of miR-185-5p enhanced autophagy and alleviated II/R intestinal barrier damage by targeting ATG101.

Chemotherapeutic drug resistance is a challenge for the effective treatment of OSCC. There are a couple of studies on the involvement of microRNAs (miRNAs) in chemoresistance of oral squamous cell carcinoma (OSCC), but the exact molecular events in many cases are not clearly understood. In this work, we intend to track down key miRNA(s) and unveil their regulatory molecular mechanisms in imparting chemoresistance in this lethal cancer. We analyzed gene and miRNA array profiles of drug-resistant OSCC cells, predicted miRNA targets, performed enrichment analysis, and validated our findings in cisplatin-sensitive and cisplatin-resistant SCC9 and H357 OSCC cells. We evaluated the anticancer and chemosensitivity roles of selected miRNA by adopting several molecular assays like qRT-PCR, MTT assay, wound healing assay, fluorescence imaging by DCFHDA, AO/EB staining, DAPI, and γ-H2AX accumulation assay. We also validated the miRNA-target binding by qRT-PCR and luciferase reporter assay. Among the enriched miRNAs, we found miR-185–5p downregulated in cisplatin-resistant OSCC cells as a signature miRNA modulating chemoresistance. The upregulation of miR-185–5p by mimic transfection restores cisplatin sensitivity by decreasing cell viability in a dose-dependent manner and increasing ROS-induced DNA damage and apoptosis. miR-185–5p overexpression increases miR-203a-3p expression through negative regulation of SOX9. siRNA-mediated silencing of the SOX9 also shows similar results. Mechanistically, miR-185–5p dependent miR-203a-3p expression decreases cisplatin efflux and cisplatin-induced DNA damage repair by regulating ABCC1, ABCB1, RRM2, and RAN. This study will pave the way for employing this miR-185–5p as a combination therapeutic strategy to combat cisplatin resistance in oral cancer. [Display omitted] • miR-185–5p is downregulated in cisplatin-resistant OSCC and correlated with drug resistance. • Overexpression of miR-185–5p increases cisplatin chemosensitivity through increased ROS production, DNA damage, and apoptosis. • miR-185–5p sensitizes cisplatin by decreasing drug efflux by restoring the expression of miR-203a-3p through negative regulation of SOX9 [ABSTRACT FROM AUTHOR]

Circular RNA (circRNA) has been confirmed to be the key regulators of diabetic nephropathy (DN) progression. However, the role of circ_0114428 in the DN progression remains unclear. Glomerular mesangial cells (GMCs) were treated with high glucose (HG) to mimic DN cell models in vitro. The expression levels of circ_0114428, microRNA (miR)-185-5p, and SMAD3 mRNA were examined by quantitative real-time PCR. Cell proliferation ability was detected by MTT assay, EdU staining and flow cytometry. The protein levels of proliferation marker, fibrosis markers, epithelial-mesenchymal transition (EMT) markers and SMAD3 were measured by western blot assay. The interaction between miR-185-5p and circ_0114428 or SMAD3 was confirmed via dual-luciferase reporter assay, RIP assay and RNA pull-down assay. Our data showed that circ_0114428 was upregulated in HG-induced GMCs. Circ_0114428 overexpression could aggravate the promotion effect of HG on the proliferation, fibrosis and EMT process of GMCs, while its knockdown had an opposite effect. In the terms of mechanisms, circ_0114428 could sponge miR-185-5p to regulate SMAD3. MiR-185-5p inhibitor could reverse the suppressive effect of circ_0114428 knockdown on the proliferation, fibrosis and EMT process in HG-induced GMCs. Also, SMAD3 overexpression abolished the inhibition of miR-185-5p on the proliferation, fibrosis and EMT process in HG-induced GMCs. Taken together, our data suggested that circ_0114428 might promote DN progression by regulating the miR-185-5p/SMAD3 axis. [ABSTRACT FROM AUTHOR]

The aim of this study is to investigate the impact of sh-LncRNA ASB16-AS1 on doxorubicin (DOX) resistance in colorectal cancer (CRC). First, an in vitro study was conducted to investigate the effects of LncRNA ASB16-AS1, miR-185-5p, and TEAD1 on drug resistance in CRC cells. Subsequently, utilizing nanotechnology, poly(beta amino esters) (PBAE)/zeolitic imidazolate framework-8 (ZIF-8)@sh-LncRNA ASB16-AS1 nanoparticles (PZSNP) were synthesized and characterized, evaluating their cellular toxicity and hemolytic activity. Finally, a mouse subcutaneous tumor model was established by subcutaneous injection of SW480/DOX cell suspension to investigate the impact of PZSNP on the tumor. Under DOX treatment, downregulation of LncRNA ASB16-AS1, overexpression of miR-185-5p, or downregulation of TEAD1 suppressed the viability and proliferation of drug-resistant CRC cells while promoting apoptosis. Conversely, overexpression of LncRNA ASB16-AS1, inhibition of miR-185-5p, or overexpression of TEAD1 enhanced the viability and proliferation of drug-resistant CRC cells while inhibiting apoptosis. The synthesized PZSNP exhibited a spherical shape with an average particle size of 123.6 nm, possessed positive charge, displayed good stability. It effectively encapsulated shRNA and displayed low cellular toxicity and hemolytic activity. Under DOX treatment, significant tumor necrosis was observed in the PZSNP group, and tumor growth was suppressed without causing weight loss. LncRNA ASB16-AS1, miR-185-5p, and TEAD1 are involved in regulating cell viability, proliferation, and apoptosis, contributing to drug resistance in CRC cells. sh - LncRNA ASB16-AS1 enhances the sensitivity of CRC cells to DOX during treatment, and in vivo delivery of PZSNP may serve as an effective strategy to overcome chemotherapy resistance in CRC.

Background Osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) is essential for bone formation, and its dysfunction is reported to be associated with osteoporosis (OP). Recent researches have determined that lncRNA PART1 participates in the pathogenesis of multiple diseases. However, its role in modulating osteogenic differentiation of hBMSCs is unclear. Methods PART1, miR-185-5p, and RUNX3 levels were assessed via RT-qPCR. The protein levels of OCN, OSN, and COL1A1 were measured by western blotting. The osteoblastic phenotype was evaluated via ALP activity and ARS staining. The relationship between miR-185-5p and PART1 or RUNX3 was validated by luciferase reporter, RIP assays. Results PART1 and RUNX3 expression were enhanced during hBMSC osteogenic differentiation. PART1 deletion decreased OCN, OSN, and COL1A1 levels and weakened ALP activity, but promoted the apoptosis of hBMSCs. Moreover, PART1 served as a ceRNA to influence the RUNX3 level via targeting miR-185-5p. In addition, RUNX3 was verified to activate the transcription of PART1 in hBMSCs. Finally, rescue assays indicated that suppression of miR-185-5p or addition of RUNX3 partially abolished the effects of PART1 knockdown on the levels of OCN, OSX, and COL1A1 levels, ALP activity, and apoptosis. Conclusion Our study elaborated that PART1/miR-185-5p/RUNX3 feedback contributed to osteogenic differentiation and inhibited the hBMSCs apoptosis, suggesting that PART1 might be a novel target for OP treatment.

The study aims to explore the molecular mechanism involved in Parkinson's disease (PD). Hsa_circ_0004381, microRNA-185-5p (miR-185-5p), and Rac family small GTPase 1 (RAC1) level were measured by real-time quantitative polymerase chain reaction (RT-qPCR). Furthermore, cell viability and apoptosis rate were assessed by Cell Counting Kit-8 (CCK-8) and flow cytometry assays, respectively. Protein levels of B cell lymphoma-2 (Bcl-2), Bcl-2-related X protein (Bax), cleaved-caspase 3 (c-caspase 3), and RAC1 were determined by western blot assay. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 were detected by enzyme-linked immunosorbent assay (ELISA). The ROS generation and LDH and SOD activity were detected by the corresponding kits. The binding relationship between miR-185-5p and hsa_circ_0004381 or RAC1 was predicted by Starbase and then verified by a dual-luciferase reporter and RNA Immunoprecipitation (RIP) assays. Hsa_circ_0004381 and RAC1 were increased, and miR-185-5p was decreased in MPP+-triggered SK-N-SH cells. Moreover, hsa_circ_0004381 silencing promoted cell viability, and repressed apoptosis, inflammatory response, and oxidative stress in MPP+-treated SK-N-SH cells. The mechanical analysis suggested that hsa_circ_0004381 served as a sponge of miR-185-5p to affect RAC1 expression. Hsa_circ_0004381 could contribute to MPP+-triggered neuron injury by targeting the miR-185-5p/RAC1 axis, which provided a novel insight into the pathogenesis and treatment of PD. [ABSTRACT FROM AUTHOR]

This article was designed to explore the effects and mechanisms of miR-185-5p on the replication of hepatitis C virus (HCV). Quantitative reverse transcription PCR (qRT-PCR) was performed for detecting the abundance of miR-185-5p and HCV RNA in HCV-infected primary hepatocytes and Huh7.5 cells. Dual-luciferase reporter gene assay was used for exploring the interaction between miR-185-5p and GALNT8. Western blot analyzed protein expression of GALNT8, NS3, and NS5A. miR-185-5p was remarkably downregulated in HCV-infected primary hepatocytes and Huh7.5 cells. miR-185-5p upregulation inhibited HCV RNA expression, while its inhibition promoted HCV replication. miR-185-5p induced accumulation of NS3 and NS5A in the cells. Dual-luciferase reporter gene assay verified the targeted relationship between miR-185-5p and GALNT8. In addition, the effects of overexpressing or knocking down miR-185-5p on HCV replication could be correspondingly eliminated by the overexpression or knockdown of GALNT8. miR-185-5p may target GALNT8 in JFH1-infected Huh7.5 cells and then inhibit HCV replication. miR-185-5p may be a potential target for treating HCV.

Abstract Background Glioma is the most common and malignant tumor of central nervous system. The tumor initiation, self-renewal, and multi-lineage differentiation abilities of glioma stem cells (GSCs) are responsible for glioma proliferation and recurrence. Although circular RNAs (circRNAs) play vital roles in the progression of glioma, the detailed mechanisms remain unknown. Methods qRT-PCR, western blotting, immunohistochemistry, and bioinformatic analysis were performed to detect the expression of circATP5B, miR-185-5p, HOXB5, and SRSF1. Patient-derived GSCs were established, and MTS, EDU, neurosphere formation, and limiting dilution assays were used to detect the proliferation of GSCs. RNA-binding protein immunoprecipitation, RNA pull-down, luciferase reporter assays, and chromatin immunoprecipitation assays were used to detect these molecules’ regulation mechanisms. Results We found circATP5B expression was significantly upregulated in GSCs and promoted the proliferation of GSCs. Mechanistically, circATP5B acted as miR-185-5p sponge to upregulate HOXB5 expression. HOXB5 was overexpressed in glioma and transcriptionally regulated IL6 expression and promoted the proliferation of GSCs via JAK2/STAT3 signaling. Moreover, RNA binding protein SRSF1 could bind to and promote circATP5B expression and regulate the proliferation of GSCs, while HOXB5 also transcriptionally regulated SRSF1 expression. Conclusions Our study identified the SRSF1/circATP5B/miR-185-5p/HOXB5 feedback loop in GSCs. This provides an effective biomarker for glioma diagnosis and prognostic evaluation.

Pyroptosis is associated with various cardiovascular diseases. Increasing evidence suggests that long noncoding RNAs (lncRNAs) have been implicated in gene regulation, but how lncRNAs participate in the regulation of pyroptosis in the heart remains largely unknown. In this study, we aimed to explore the antipyroptotic effects of lncRNA FGF9-associated factor (FAF) in acute myocardial infarction (AMI). The expression patterns of lncRNA FAF, miR-185-5p and P21 activated kinase 2 (PAK2) were detected in hypoxia/ischaemia-induced cardiomyocytes. Hoechst 33342/PI staining, lactate dehydrogenase (LDH) release assay, immunofluorescence and Western blotting were conducted to assay cell pyroptosis. The interaction between lncRNA FAF, miR-185-5p and PAK2 was verified by bioinformatics analysis, small RNA sequencing luciferase reporter assay and qRT-PCR. The expression of LncRNA FAF was downregulated in hypoxic cardiomyocytes and myocardial tissues. Overexpression of lncRNA FAF could attenuate cardiomyocyte pyroptosis, improve cell viability and reduce infarct size during the procession of AMI. Moreover, lncRNA FAF was confirmed as a sponge of miR-185-5p and promoted PAK2 expression in cardiomyocytes. Collectively, our findings reveal a novel lncRNA FAF/miR-185-5p/PAK2 axis as a crucial regulator in cardiomyocyte pyroptosis, which might be a potential therapeutic target of AMI. [ABSTRACT FROM AUTHOR]

Long non-coding RNAs (lncRNAs) are crucial players in tumour progression. Herein, this work was designated to decipher the clinical significance, function and molecular mechanism of a lncRNA, differentiation antagonizing non-coding RNA (DANCR) in colorectal cancer (CRC). Quantitative real-time polymerase chain reaction was adopted to examine DANCR, miR-185-5p and HMGA2 mRNA expressions in CRC tissues and cells. Both gain-of-function and loss-of-function cell models for DANCR were established, and then MTT, wound healing and Transwell, flow cytometry assays were carried out to detect the proliferation, migration, invasion, cell cycle and apoptosis of CRC cells. Dual-luciferase reporter gene assay and RIP assay were utilized to validate the targeting relationships between DANCR and miR-185-5p. Western blot was employed for detecting high mobility group A2 (HMGA2) expressions in CRC cells. In this study, we demonstrated that the expression of DANCR was elevated in CRC tissues and cell lines, and its high expression was significantly associated with increased TNM stage and positive lymph node metastasis. DANCR overexpression promoted CRC cell proliferation, migration, invasion and cell cycle progression, but inhibited apoptosis; while knocking down DANCR caused the opposite effects. DANCR was further identified as a molecular sponge for miR-185-5p, and DANCR could indirectly increase the expression of HMGA2 via repressing miR-185-5p. In conclusion, DANCR/miR-185-5p/HMGA2 axis participated in the progression of CRC. [ABSTRACT FROM AUTHOR]

Ischemic stroke is a common disease threatening human health. ADAMTS9-AS2 is a lncRNA that has been widely studied in tumors, but not in ischemic stroke. The purpose of this study was to investigate the role and potential molecular mechanism of ADAMTS9-AS2 in endothelial cell function after ischemic stroke in vivo and in vitro. The results showed that ADAMTS9-AS2 was decreased in the plasma of acute ischemic stroke (AIS) patients and in the brain tissue and plasma of MCAO mice, and the low expression of ADAMTS9-AS2 was associated with the increase in infarct size. Besides, compared with the control group, MCAO treatment slightly promoted angiogenesis, which was enhanced by the overexpression of ADAMTS9-AS2. Molecular mechanism results indicated that ADAMTS9-AS2 and miR-185-5p affected the pathological process of ischemic stroke through ceRNA mechanism, and IGFBP-2 was a downstream target gene of miR-185-5p. These findings demonstrated that ADAMTS9-AS2 promoted angiogenesis through regulating miR-185-5p/IGFBP-2 axis, suggesting that ADAMTS9-AS2 may be a potential marker for the diagnosis and treatment of neurological diseases. [ABSTRACT FROM AUTHOR]

Osteosarcoma (OS) is a malignant bone tumor characterized by poor prognosis due to its regional invasion and early metastasis. In this study, we aimed to find the role and the underlying mechanism of Cathepsin E (CTSE) in OS growth and metastasis. We found CTSE is upregulated in metastatic OS, rather than in the primary lesion, as confirmed by RT-qPCR and western blot analysis of clinical OS samples. Furthermore, both in vitro and in vivo experiments illustrated that CTSE promoted both growth and metastasis of OS cells, partially mediated through the modulation of Epithelial–Mesenchymal Transition (EMT). Bioinformatics analysis predicted that miR-185-5p downregulates CTSE via directly binding to the 3'UTR of CTSE, which was verified by luciferase reporter assay and rescue assays. This study reported for the first time that CTSE is a potential biomarker in OS tumorigenesis and metastasis, providing a promising therapeutic target for OS treatment. [ABSTRACT FROM AUTHOR]

The involvement of certain circular RNAs (circRNAs) in the development of glioma has been revealed. CircRNA periostin (circPOSTN) was validated to be positively associated with glioma cell growth and metastasis. However, the mechanism underlying circPOSTN in glioma tumorigenesis remain vague. The expression of circPOSTN, KIF1B (Kinesin Family Member 1B) and miR-185–5p was detected using quantitative real-time polymerase chain reaction and Western blot. In vitro assays were conducted using cell counting kit-8 assay, colony formation assay, EdU assay, flow cytometry, Western blot, and transwell assay, respectively. The direct interactions between miR-185–5p and circPOSTN or KIF1B was confirmed by using dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. CircPOSTN was highly expressed in glioma tissues and cells. Knockdown of circPOSTN restrained glioma cell proliferation, migration and invasion in vitro, as well as hindered glioma xenograft growth in vivo. Mechanistically, circPOSTN acted as miR-185–5p sponge to up-regulate the expression of its target KIF1B. Moreover, miR-185–5p inhibition reversed the anticancer effects of circPOSTN knockdown on glioma tumorigenesis, and miR-185–5p re-expression suppressed the malignant phenotype of glioma cells via targeting KIF1B. CircPOSTN acted as an oncogene to expedite glioma tumorigenesis via targeting miR-185–5p/KIF1B axis, indicating a potential therapeutic target for glioma. • CircPOSTN was highly expressed in glioma tissues and cells. • Knockdown of circPOSTN restrained glioma cell proliferation and mobility in vitro, as well as impeded tumor growth in vivo. • CircPOSTN served as a sponge for miR-185–5p to positively modulate KIF1B expression. • CircPOSTN contributed to glioma tumorigenesis via miR-185–5p/KIF1B axis. [ABSTRACT FROM AUTHOR]

The emerging role of long noncoding RNAs (lncRNAs) in cancer, especially in lung adenocarcinoma (LUAD), is attracting increasingly more attention as a potential therapeutic target. However, whether lncRNA LINC00205 regulates the malignancy of LUAD has not been characterized. In this study, we discovered that LINC00205 was markedly upregulated in LUAD tissues and cell lines and correlated with poor prognosis of patients with LUAD. Our data showed that LINC00205 promoted the migration and proliferation of LUAD cells in vitro and tumor growth in vivo. Notably, the tumor suppressor miR-185-5p was found to be a direct target of LINC00205. In addition, miR-185-5p diminished the promotion of cell proliferation and migration mediated by LINC00205, whereas miR-185-5p inhibition had the opposite effect. In summary, our results show that LINC00205 contributes to LUAD malignancy by sponging miR-185-5p, which provides new insight into LUAD progression. [ABSTRACT FROM AUTHOR]

As a common malignant tumor, colorectal cancer (CRC) has a high incidence. Recent investigations have suggested that although great improvement has been achieved in the survival rate of early-stage CRC patients, the overall survival rate remains low. Mounting reports have proved that lncRNAs take part in the development of various cancers and possess the regulatory functions in cancers. For example, ASB16 antisense RNA 1 (ASB16-AS1) is a poorly researched novel lncRNA whose specific functions in CRC are still unknown. In our research, we discovered that ASB16-AS1 was with high expression in CRC cells. In addition, ASB16-AS1 silencing restrained the proliferation, migration, invasion, and stemness while accelerating cell apoptosis of CRC cells. Mechanism experiments were applied to explore the regulatory mechanism of ASB16-AS1. It turned out that miR-185-5p could interact with ASB16-AS1 and inhibited the progression of CRC cells. TEAD1 (TEA domain transcription factor1) - a major effector of the Hippo signaling was proved to serve as the target of miR-185-5p and promote CRC development. In short, ASB16-AS1 drove the progression of CRC through the regulation of miR-185-5p/TEAD1 axis. [ABSTRACT FROM AUTHOR]

Osteosarcoma (OS) is a common malignant tumor in the world. Circular RNAs are endogenous non-coding RNAs that have been linked to the development of cancer. However, the role of circ_001569 in OS progression is still unclear. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of circ_001569, microRNA-185-5p (miR-185-5p) and flotillin-2 (FLOT2). The abilities of cell proliferation, migration and invasion were evaluated by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and Transwell assays, respectively. Also, western blot analysis was performed to assess the levels of epithelial–mesenchymal transition (EMT)-related proteins and FLOT2 protein. Besides, the dual-luciferase reporter assay was used to verify the interactions among circ_001569, miR-185-5p and FLOT2. Circ_001569 expression was increased in OS tissues and cells, and its knockdown reduced the proliferation, migration, invasion and EMT of OS cells. MiR-185-5p could interact with circ_001569. Inhibition of miR-185-5p could recover the suppression effects of silenced-circ_001569 on the proliferation and metastasis of OS cells. Furthermore, FLOT2 was a target of miR-185-5p. Overexpressed FLOT2 could restore the inhibition effects of miR-185-5p mimic on the proliferation and metastasis of OS cells. Also, FLOT2 expression was regulated by circ_001569 and miR-185-5p. In addition, circ_001569 knockdown also reduced the OS tumor growth in vivo. Circ_001569 might act as an oncogene in OS progression by regulating the miR-185-5p/FLOT2 axis, which provided a reliable new approach for the treatment of OS patients.

Esophageal squamous cell carcinoma (ESCC) is a common cancer occurring in males and females worldwide. Accumulating evidence continues to highlight the crucial roles of long non-coding RNAs (lncRNAs) in the process of tumorigenesis. However, the regulatory mechanism of lncRNAs in ESCC remains unclear. The aim of this study is to elucidate the role of lncRNA Krüppel-like factor 3 antisense RNA 1 (KLF3-AS1) in ESCC by regulating miR-185-5p and KLF3. Initially, ESCC cell spheres with stem cell-like properties were prepared by suspension culture, and subsequently characterized by assessing colony formation ability and stem cell markers. LncRNA KLF3-AS1 was found to be poorly expressed in ESCC and could upregulate the expression of KLF3 by binding to miR-185-5p. lncRNA KLF3-AS1 upregulation was observed to inhibit miR-185-5p, thereby contributing to decreased expression of SOX2 and Oct4 (octamer-binding transcription factor 4). Furthermore, enhancement of lncRNA KLF3-AS1 resulted in reduced colony formation ability, cell invasion and migration, and tumor volume in vivo while promoting cell apoptosis in ESCC through downregulation of miR-185-5p. Collectively, this study indicated that lncRNA KLF3-AS1 inhibited ESCC cell invasion and migration by impairing miR-185-5p-mediated inhibition of KLF3, highlighting a promising novel potential target for ESCC treatment.

Abstract Background Breast cancer is the most common cancer types among women. Recent researches have focused on determining the efficiency of alternative molecules and miRNAs in breast cancer treatment. The aim of this study was to determine the effect of usnic acid response-miR-185-5p on proliferation in the breast cancer cell and to determine its relationship with apoptosis pathway. Methods The cell proliferation and cell apoptosis rate were significantly increased following the ectopic expression of miR-185-5p in BT-474 cells. Furthermore, the results of cell cycle assay performed by flow cytometry revealed that the transfection with miR-185-5p induced G1/S phase arrest. The apoptosis-related genes expression analysis was performed by qRT-PCR and the direct target of miR-185-5p in BT-474 cells was identified by western blot and luciferase reporter assay. Results Our data showed that miR-185-5p can cause significant changes in apoptosis-related genes expression levels, suggesting that cell proliferation was suppressed by miR-185-5p via inducing apoptosis in breast cancer cells. According to western blot results, miR-185-5p lead to decrease BCL2 protein level in BT-474 cells and direct target of miR-185-5p was identified as BCL by luciferase reporter assay. Conclusion This study revealed that miR-185-5p may be an effective agent in the treatment of breast cancer.

APP misexpression plays a crucial role in triggering a complex pathological cascade, leading to Alzheimer’s disease (AD). But how the expression of APP is regulated in pathological conditions remains poorly understood. In this study, we found that the exosomes isolated from AD mouse brain promoted APP expression in neuronal N2a cells. Moreover, exosomes derived from N2a cells with ectopic expression of APP (APP-EXO) also induced APP dysregulation in normal N2a cells. Surprisingly, the effects of APP-EXO on APP expression in recipient cells were not mediated by the direct transferring of APP gene products. Instead, the effects of APP-EXO were highly likely mediated by the reduction of the expression levels of exosomal miR-185-5p. We found that the 3′UTR of APP transcripts binds to miR-185-5p, therefore inhibiting the sorting of miR-185-5p to exosomes. N2a cell-derived exosomes with less amount of miR-185-5p exert similar roles in APP expression to APP-EXO. Lastly, we demonstrated a significant decline of serum exosomal miR-185-5p in AD patients and AD mice, versus the corresponding controls. Together, our results demonstrate a novel mechanism in the exosome-dependent regulation of APP, implying exosomes and exosomal miRNAs as potential therapeutic targets and biomarkers for AD treatment and diagnosis, respectively.

Background: There are ever increasing researches implying that noncoded RNAs (ncRNAs) specifically circular RNAs (circRNAs) and microRNAs (miRNAs) in exosomes play vital roles in respiratory disease. However, the detailed mechanisms persist to be unclear in mycobacterial infection. Methods: In order to detect circRNAs and miRNAs expression pattern and potential biological function in tuberculosis, we performed immense parallel sequencing for exosomal ncRNAs from THP-1-derived macrophages infected by Mycobacterium tuberculosis H37Ra, Mycobacterium bovis BCG and control Streptococcus pneumonia , respectively and uninfected normal cells. Besides, THP-1-derived macrophages were used to verify the validation of differential miRNAs, and monocytes from PBMCs and clinical plasma samples were used to further validate differentially expressed miR-185-5p. Results: Many exosomal circRNAs and miRNAs associated with tuberculosis infection were recognized. Extensive enrichment analyses were performed to illustrate the major effects of altered ncRNAs expression. Moreover, the miRNA–mRNA and circRNA–miRNA networks were created and expected to reveal their interrelationship. Further, significant differentially expressed miRNAs based on Exo-BCG, Exo-Ra and Exo-Control, were evaluated, and the potential target mRNAs and function were analyzed. Eventually, miR-185-5p was collected as a promising potential biomarker for tuberculosis. Conclusion: Our findings provide a new vision for exploring biological functions of ncRNAs in mycobacterial infection and screening novel potential biomarkers. To sum up, exosomal ncRNAs might represent useful functional biomarkers in tuberculosis pathogenesis and diagnosis. [ABSTRACT FROM AUTHOR]

Osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) is essential for bone formation, and its dysfunction is reported to be associated with osteoporosis (OP). Recent researches have determined that lncRNA PART1 participates in the pathogenesis of multiple diseases. However, its role in modulating osteogenic differentiation of hBMSCs is unclear. PART1, miR-185-5p, and RUNX3 levels were assessed via RT-qPCR. The protein levels of OCN, OSN, and COL1A1 were measured by western blotting. The osteoblastic phenotype was evaluated via ALP activity and ARS staining. The relationship between miR-185-5p and PART1 or RUNX3 was validated by luciferase reporter, RIP assays. PART1 and RUNX3 expression were enhanced during hBMSC osteogenic differentiation. PART1 deletion decreased OCN, OSN, and COL1A1 levels and weakened ALP activity, but promoted the apoptosis of hBMSCs. Moreover, PART1 served as a ceRNA to influence the RUNX3 level via targeting miR-185-5p. In addition, RUNX3 was verified to activate the transcription of PART1 in hBMSCs. Finally, rescue assays indicated that suppression of miR-185-5p or addition of RUNX3 partially abolished the effects of PART1 knockdown on the levels of OCN, OSX, and COL1A1 levels, ALP activity, and apoptosis. Our study elaborated that PART1/miR-185-5p/RUNX3 feedback contributed to osteogenic differentiation and inhibited the hBMSCs apoptosis, suggesting that PART1 might be a novel target for OP treatment. [ABSTRACT FROM AUTHOR]

Circular RNAs (circRNAs) have been reported to play vital roles in atherosclerosis. However, the precise roles of circUBR4 in atherosclerosis remain unclear. The purpose of this study is to investigate the regulatory roles of circUBR4 in atherosclerosis. The expression levels of circUBR4, miR-185-5p, and Fibroblast growth factor receptor substrate 2 (FRS2) were analyzed by real-time quantitative polymerase chain reaction (RT-qPCR) assay. Human vascular smooth muscle cells (VSMCs) were treated with oxidized low-density lipoprotein (ox-LDL) to mimic atherosclerosis condition in vitro. Cell proliferation was determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazol-3-ium bromide (MTT), colony-forming, and 5-ethynyl-2′-deoxyuridine (EdU) assays. Wound healing and transwell assays were used to assess cell migration. The interaction relationship between miR-185-5p and circUBR4 or FRS2 was confirmed by dual-luciferase reporter and RNA pull-down assays. CircUBR4 was overexpressed in atherosclerosis patients and VSMCs treated with ox-LDL, and the knockdown of circUBR4 abolished ox-LDL-induced enhanced effects on the proliferation and migration of VSMCs. MiR-185-5p, interacted with FRS2, was a target of circUBR4 in VSMCs. The silencing of miR-185-5p reversed the effects caused by circUBR4 knockdown on ox-LDL-induced VSMCs. In addition, overexpression of miR-185-5p suppressed the proliferation and migration of VSMCs by targeting FRS2. CircUBR4 contributed to ox-LDL-induced VSMC proliferation and migration through up-regulating FRS2 via miR-185-5p. [ABSTRACT FROM AUTHOR]

AT-rich interaction domain 1A (ARID1A) is a tumor suppressor gene that mutates in several cancer types, including breast cancer, ovarian cancer, and colorectal cancer (CRC). In colon adenocarcinoma (COAD), the low expression of ARID1A was reported but the molecular reason is unclear. We noticed that ARID1A low expression was associated with increased levels of miR-185 in the COAD. Therefore, this study aims to explore ncRNA-dependent mechanism that regulates ARID1A expression in COAD regarding miR-185. The expression of ARID1A was tested in COAD cell line under the effect of miR-185 mimics compared with inhibitor. The molecular features associated with loss of ARID1A and its association with tumor prognosis were analyzed using multi-platform data from The Cancer Genome Atlas (TCGA), and gene set enrichment analysis (GSEA) to identify potential signaling pathways associated with ARID1A alterations in colon cancer. Kaplan-Meier survival curve showed that a low level of ARID1A was closely related to low survival rate in patients with COAD. Results showed that inhibiting miR-185 expression in the COAD cell line significantly restored the expression of ARID1A. Further, the increased expression of ARID1A significantly improved the prolonged overall survival of COAD. We noticed that there is a possible relationship between ARID1A high expression and tumor microenvironment infiltrating immune cells. Furthermore, the increase of ARID1A in tumor cells enhanced the response of inflammatory chemokines. In conclusion, this study demonstrates that ARID1A is a direct target of miR-185 in COAD that regulates the immune modulations in the microenvironment of COAD.

AT-rich interaction domain 1A (ARID1A) is a tumor suppressor gene that mutates in several cancer types, including breast cancer, ovarian cancer, and colorectal cancer (CRC). In colon adenocarcinoma (COAD), the low expression of ARID1A was reported but the molecular reason is unclear. We noticed that ARID1A low expression was associated with increased levels of miR-185 in the COAD. Therefore, this study aims to explore ncRNA-dependent mechanism that regulates ARID1A expression in COAD regarding miR-185. The expression of ARID1A was tested in COAD cell line under the effect of miR-185 mimics compared with inhibitor. The molecular features associated with loss of ARID1A and its association with tumor prognosis were analyzed using multi-platform data from The Cancer Genome Atlas (TCGA), and gene set enrichment analysis (GSEA) to identify potential signaling pathways associated with ARID1A alterations in colon cancer. Kaplan-Meier survival curve showed that a low level of ARID1A was closely related to low survival rate in patients with COAD. Results showed that inhibiting miR-185 expression in the COAD cell line significantly restored the expression of ARID1A. Further, the increased expression of ARID1A significantly improved the prolonged overall survival of COAD. We noticed that there is a possible relationship between ARID1A high expression and tumor microenvironment infiltrating immune cells. Furthermore, the increase of ARID1A in tumor cells enhanced the response of inflammatory chemokines. In conclusion, this study demonstrates that ARID1A is a direct target of miR-185 in COAD that regulates the immune modulations in the microenvironment of COAD. [ABSTRACT FROM AUTHOR]

Background: Long non-coding RNA differentiation antagonizing non-protein coding RNA (DANCR) acts as an oncogene in different cancers, although its roles in prostate cancer are not fully reported. We aimed to explore its mechanism in facilitating the malignancy of prostate cancer. Methods: The expression of DANCR, microRNA (miR)-185-5p and LIM and SH3 protein 1 (LASP1) in 40 pairs of prostate cancer tissues and normal tissues, five prostate cancer cell lines and one epithelial cell line was assessed by a quantitative real-time polymerase chain reaction, western blotting and immunohistochemistry, respectively. In transfected PC3 and C4-2 cells, cell proliferation, migration, invasion, cell cycle distribution and epithelial-mesenchymal transition (EMT) protein expression were tested via cell counting kit-8, wound healing, transwell, flow cytometry and western blot assays, respectively. The interactions between DANCR, miR-185-5p and LASP1 were verified by a dual-luciferase reporter assay. Rescue experiments were conducted to determine the roles of DANCR on the malignant properties of PC3 and C4-2 cells. The involvement of the signaling pathway was examined using a p-FAK inhibitor. Results: DANCR and LASP1 expression was enhanced, whereas miR-185-5p expression was diminished in prostate cancer tissues and cell lines. Knockdown of DANCR suppressed cell proliferation, migration, invasion, G1-S transition and expression of EMT proteins of the transfected PC3 and C4-2 cells. DANCR sponged miR-185-5p to upregulate LASP1 expression. DANCR-miR-185-5p-LASP1 axis activates the FAK/PI3K/AKT/GSK3ß/Snail pathway to promote the malignant properties of PC3 and C4-2 cells. Conclusions: These findings suggest that DANCR exerts oncogenic roles in prostate cancer via the miR-185-5p/LASP1 axis activating the FAK/PI3K/AKT/GSK3ß/Snail pathway. It can be a potential biomarker in the diagnosis and monitoring of prostate cancer. [ABSTRACT FROM AUTHOR]