TargetScan Research Articles

Astrometric accuracy of snapshot fast radio burst localisations with ASKAP

Abstract The recent increase in well-localised fast radio bursts (FRBs) has facilitated in-depth studies of global FRB host properties, the source circumburst medium, and the potential impacts of these environments on the burst properties. The Australian Square Kilometre Array Pathfinder (ASKAP) has localised 11 FRBs with sub-arcsecond to arcsecond precision, leading to sub-galaxy localisation regions in some cases and those covering much of the host galaxy in others. The method used to astrometrically register the FRB image frame for ASKAP, in order to align it with images taken at other wavelengths, is currently limited by the brightness of continuum sources detected in the short-duration (‘snapshot’) voltage data captured by the Commensal Real-Time ASKAP Fast Transients (CRAFT) software correlator, which are used to correct for any frame offsets due to imperfect calibration solutions and estimate the accuracy of any required correction. In this paper, we use dedicated observations of bright, compact radio sources in the low- and mid-frequency bands observable by ASKAP to investigate the typical astrometric accuracy of the positions obtained using this so-called ‘snapshot’ technique. Having captured these data with both the CRAFT software and ASKAP hardware correlators, we also compare the offset distributions obtained from both data products to estimate a typical offset between the image frames resulting from the differing processing paths, laying the groundwork for future use of the longer duration, higher signal-to-noise ratio (S/N) data recorded by the hardware correlator. We find typical offsets between the two frames of ${\sim}0.6$ and ${\sim}0.3$ arcsec in the low- and mid-band data, respectively, for both RA and Dec. We also find reasonable agreement between our offset distributions and those of the published FRBs. We detect only a weak dependence in positional offset on the relative separation in time and elevation between target and calibrator scans, with the trends being more pronounced in the low-band data and in Dec. Conversely, the offsets show a clear dependence on frequency in the low band, which we compare to the frequency-dependent Dec. offsets found in FRB 200430. In addition, we present a refined methodology for estimating the overall astrometric accuracy of CRAFT FRBs.

MiRNA-191-5p represses cell growth by targeting CDK6 in gastric cancer

Objective: To investigate the effects of miR-191-5p on cell migration, clone formation and proliferation of gastric cancer (GC) cells. Methods: The level of miR-191-5p expression was detected by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) in 60 paired GC tissues and their adjacent normal tissues. miR-191-5p overexpression was achieved by transfection of construct pcDNA-miR-191-5p into GC cells. The migration, clone formation and proliferation of GC cells were detected by the scratch wound assay, clone formation assay and cell counting kit-8 (CCK-8), respectively. Low expression of miR-191-5p was achieved with miRNA-191-5p inhibitor. The binding sites of cyclin-dependent kinase 6 (CDK6) and miR-191-5p were analyzed using TargetScan software, and the interaction of CDK6 and miR-191-5p was verified using dual-fluorescence reporter gene expression. Western blot (WB) was used to detect the effect of miR-191-5p on the expression of p21 and CDK6 proteins. Results: miR-191-5p decreased in 53 cases (88%) of GC tissues compared to their controls. Furthermore, overexpression of miR-191-5p effectively inhibited the migration, clone formation and proliferation of GC cells (P<0.05). Dual-fluorescence reporter confirmed that miR-191-5p bound to 3'UTR of CDK6. WB showed that pcDNA-miR-191-5p inhibited the CDK6 expression but promoted the p21. Conclusion: Down-regulation of miR-191-5p has a correlation with the progression of GC. Overexpression of miR-191-5p can decrease the expression of CDK6 and inhibit the growth of GC cells.

Reverberation-based high-speed guided-wave ultrasonic propagation imager for structural inspection of thick composites

Increasing use of thick composite structures in many fields, including the aerospace and shipbuilding industries, necessitates appropriate inspection techniques to ensure structural integrity. Of many nondestructive testing techniques, laser ultrasonic testing is a non-contact method that yields accurate inspection results with high spatial resolution; this method was adopted in this study for the inspection of thick composite structures. We propose a novel guided-wave ultrasonic propagation imaging (UPI) method, called the reverberation wave UPI (R-wave UPI), which is capable of faster target scanning and clearer highlighting of confining waves. To prove the merits of the R-wave UPI, a relatively thin carbon/epoxy composite (thickness, 12 mm) was first tested using the proposed method and the conventional through-transmission UPI. The R-wave UPI modifies the guided-wave UPI with a faster pulse repetition frequency (PRF) laser and ultrasonic energy mapping with straightforward parameter determination. The through-transmission UPI consisting of a raster mechanical scanner, a Q-switched laser, and a laser Doppler vibrometer, was used as a reference, whereas the R-wave UPI consisting of an angular laser mirror scanner, a Q-switched laser, and a PZT sensor was operated at a high PRF in the kilohertz regime to induce reverberation. The reverberation phenomenon was used in the post-image processing, called ultrasonic energy mapping (UEM). UEM with a high PRF was then compared to the through-transmission inspection reference to verify its accuracy and reliability for mapping of structural anomalies. Finally, a thick and curved composite blade with a rough and curved surface that could not be inspected using the conventional through-transmission UPI was tested using 7 kHz PRF and its delamination was successfully visualized using the UEM.

Abstract 16596: Immune Evasion by Human Neonatal Cardiac Mesenchymal Stem Cells Demonstrates Therapeutic Application in Myocardial Infarction

Immune rejection of transplanted stem cells is a major stumbling block in designing effective therapy for myocardial infarction. Human neonatal cardiac mesenchymal stem cells (nMSCs) showed superior cardiac functional recovery compared to adult MSCs in immune competent rat MI model. However, molecular mechanisms underlying immune evasion by transplanted nMSCs in the infarcted myocardium remain unexplored. In this investigation, we demonstrate for the first-time the expression, regulation and function of CD47 in human nMSCs and its novel mechanism of immune evasion increases its regenerative potential in rat MI model. Transplanted nMSCs showed significant increased cell retention, reduced phagocytosis and CD68 + cells compared to aMSCs in rat MI model. Comparative proteomic analysis by LC-MS/MS on nMSCs and aMSCs showed that CD47 higher in nMSCs. Increased CD47 expression in nMSCs inhibited phagocytosis compared to aMSCs in vitro and in vivo . Further, CD47 blockade in nMSCs using anti-CD47, siRNA and shRNA lentiviral based approaches increased in vitro , in vivo phagocytosis, CD68 + cells and reduced cell retention and MI recovery in vivo . Microarray analysis and validation showed miR-34a was significantly higher in aMSCs than nMSCs. To unravel CD47 regulation in aMSCs, we performed target scan analysis that predicted miR-34a binds on CD47. Further, miR-34a over expression in nMSCs reduced CD47 expression, increased in vitro and in vivo phagocytosis and reduced cell retention and MI recovery. In conclusion, increased CD47 expression in nMSCs inhibit phagocytosis by CD47/SIRPα immune regulatory axis to demonstrates its immune evasion potential and its therapeutic applications in myocardial infarction.

Identification of Novel Hypothalamic MicroRNAs as Promising Therapeutics for SARS-CoV-2 by Regulating ACE2 and TMPRSS2 Expression: An In Silico Analysis.

Background: Neuroinvasion of severe acute respiratory syndrome coronavirus (SARS-CoV) is well documented and, given the similarities between this virus and SARS-CoV-2, it seems that the neurological impairment that is associated with coronavirus disease 2019 (COVID-19) is due to SARS-CoV-2 neuroinvasion. Hypothalamic circuits are exposed to the entry of the virus via the olfactory bulb and interact centrally with crucial respiratory nuclei. Hypothalamic microRNAs are considered as potential biomarkers and modulators for various diseases and future therapeutic targets. The present study aims to investigate the microRNAs that regulate the expression of hypothalamic angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2), essential elements for SARS-CoV-2 cell entry. Methods: To determine potential hypothalamic miRNAs that can directly bind to ACE2 and TMPRSS2, multiple target bioinformatics prediction algorithms were used, including miRBase, Target scan, and miRWalk2.029. Results: Our in silico analysis has revealed that, although there are over 5000 hypothalamic miRNAs, around 31 miRNAs and 29 miRNAs have shown binding sites and strong binding capacity against ACE2 and TMPRSS2, respectively. Conclusion: These novel potential hypothalamic miRNAs can be used to identify new therapeutic targets to treat neurological symptoms in COVID-19 patients via regulation of ACE2 and TMPRSS2 expression.

MiR103a-3p and miR107 are related to adaptive coping in a cluster of fibromyalgia patients.

BackgroundMicroRNA (miRNA) mainly inhibit post-transcriptional gene expression of specific targets and may modulate disease severity.ObjectiveWe aimed to identify miRNA signatures distinguishing patient clusters with fibromyalgia syndrome (FMS).Subjects and methodsWe previously determined four FMS patient clusters labelled “maladaptive”, “adaptive”, “vulnerable”, and “resilient”. Here, we cluster-wise assessed relative gene expression of miR103a-3p, miR107, miR130a-3p, and miR125a-5p in white blood cell (WBC) RNA of 31 FMS patients and 16 healthy controls. Sum scores of pain-, stress-, and resilience-related questionnaires were correlated with miRNA relative gene expression. A cluster-specific speculative model of a miRNA-mediated regulatory cycle was proposed, and its potential targets verified by the online tool “target scan human”.ResultsOne-way ANOVA revealed lower gene expression of miR103a-3p, miR107, and miR130a-3p in FMS patients compared to controls (p < 0.05). Follow-up post-hoc tests indicated the highest peak of gene expression of miR103a-3p for the adaptive cluster (p < 0.05), i.e. in patients with low disability in all symptom categories. Gene expression of miR103a-3p correlated with FMS related disability and miR107 with the score “physical abuse” of the trauma questionnaire (p < 0.05). Target scan identified sucrose non-fermentable serine/threonine protein kinase, nuclear factor kappa-b, cyclin dependent kinase, and toll-like receptor 4 as genetic targets of the miR103a/107 miRNA family.ConclusionWe show an association between upregulated gene expression of miR103a, tendentially of miR107, and adaptive coping in FMS patients. Validation of this pair of miRNA may enable to identify a somatic resilience factor in FMS.

Long noncoding RNA ZFPM2-AS1 acts as a miRNA sponge and promotes cell invasion through regulation of miR-139/GDF10 in hepatocellular carcinoma

BackgroundEmerging evidence has shown that dysregulated expression of long noncoding RNAs (lncRNAs) is implicated in liver hepatocellular carcinoma (HCC). However, the role and molecular mechanism of differentially expressed lncRNAs in HCC has not been fully explained.MethodsThe expression profiles of lncRNAs in HCC samples were derived from microarrays analysis or downloaded from The Cancer Genome Atlas (TCGA), and their correlation with prognosis and clinical characteristics were further analyzed. Silencing of lncRNA ZFPM2-AS1 was conducted to assess the effect of ZFPM2-AS1 in vitro. The miRcode and Target Scan databases were used to determine the lncRNA-miRNA-mRNA interactions. The biological functions were demonstrated by luciferase reporter assay, western blotting, PCR and rescue experiments.ResultsThe expression level of lncRNA ZFPM2-AS1 was significantly higher in HCC tissues than in adjacent normal tissues, and higher ZFPM2-AS1 was remarkably related to poor survival. Functionally, silencing of lncRNA ZFPM2-AS1 inhibited cell proliferation, migration, invasion and promoted cell apoptosis in vitro. Bioinformatics analysis based on the miRcode and TargetScan databases showed that lncRNA ZFPM2-AS1 regulated GDF10 expression by competitively binding to miR-139. miR-139 and downregulated GDF10 reversed cell phenotypes caused by lncRNA ZFPM2-AS1 by rescue analysis.ConclusionsZFPM2-AS1, an upregulated lncRNA in HCC, was associated with malignant tumor phenotypes and worse patient survival. ZFPM2-AS1 regulated the progression of HCC by acting as a competing endogenous RNA (ceRNA) to competitively bind to miR-139 and regulate GDF10 expression. Our study provides new insight into the posttranscriptional regulation mechanism of lncRNA ZFPM2-AS1 and suggests that ZFPM2-AS1/miR-139/GDF10 may act as a potential therapeutic target and prognostic biomarker for HCC.

MiR-23a induced the activation of CDC42/PAK1 pathway and cell cycle arrest in human cov434 cells by targeting FGD4

BackgroundMiRNAs play important roles in the development of ovarian cancer, activation of primitive follicles, follicular development, oocyte maturation and ovulation. In the present study, we investigated the specific role of miR-23a in cov434 cells.ResultsDownregulation of miR-23a was observed in serum of PCOS patients compared with the healthy control, suggesting the inhibitory effect of miR-23a in PCOS. MiR-23a was positively correlated with Body Mass Index (BMI) and negatively correlated with Luteinizing hormone (LH), Testostrone (T), Glucose (Glu) and Insulin (INS) of PCOS patients. MiR-23a mimic inhibited the proliferation and promoted apoptosis of human cov434 cells. In addition, flow cytometry assay confirmed that miR-23a blocked cell cycle on G0/G1 phase. MiR-23a inhibitor showed opposite results. Furthermore, double luciferase reporter assay proved that miR-23a could bind to the 3’UTR of FGD4 directly through sites predicted on Target Scan. FGD4 level was significantly suppressed by miR-23a mimic, but was significantly enhanced by miR-23a inhibitor. We further proved that miR-23a increased the expression of activated CDC42 (GTP bround) and p-PAK-1, suggesting that miR-23a induced cell cycle arrest through CDC42/PAK1 pathway.ConclusionsIn conclusion, our study reveals that miR-23a participates in the regulation of proliferation and apoptosis of cov434 cells through target FGD4, and may play a role in the pathophysiology of PCOS.

P1747GLOBAL MIRNOME PROFILE REVEALS INVOLMENT OF MITOGEN ACTIVATED PROTEIN KINASE (MAPK) PATHWAY IN PATIENTS WITH ANTIBODY MEDIATED REJECTION (CABMR) OF RENAL TRANSPLANT

Abstract Background and Aims MicroRNAs (miRNAs) are non-coding RNA that play pivotal role in modulating expression of multiple target genes at post-transcriptional level and have potential to modulate physiological &amp; pathological processes thus can be used as potential therapeutic targets. The impact of miRNA regulations have been shown in various kidney disorders but its involvement in allograft function and immunity has not been investigated thoroughly thus we studied the miRNAs expression involved in CABMR. The impact of miRNA regulation involved in allograft function and immunity has not been investigated thoroughly thus we studied the miRNAs expression involved in CABMR. Method Patients with CABMR (Banff’s classification- 2017) were included. 25 blood donors (M=18) with a mean age of 52.4Y were served as controls. Samples were processed for RNA isolation. RNA integrity was checked by running over 1% agarose gel and quantity and qualities were checked using nanodrop with 260/230 and 260/280 ≥ 1.8. Massive parallel sequencing was performed by HiSeq 3000 sequencing system (Illumina Inc.,USA). The data obtained from the sequencing run were processed and filtered using the miRNA analysis app (Illumina Inc., USA) and analysis were performed using the BaseSpace Sequence Hub genomics computing environment (Illumina Inc., USA). Dysregulated miRNAs were detected by quantitative miRNA stem loop RT-PCR technology (TaqMan MicroRNA Assays, Applied Biosystems, CA) as per manufactures protocol. Expression was measured by calculating fold change. Targets of dysregulated miRNAs were predicted by computational analysis using bioinformatics tools like miRTarBase and Target Scan Human as well as using databases. Pathway analysis of predicted targets were performed using DIANA - mirPath online analysis tool and Kyoto Encyclopedia Genes and Genomes (KEGG) database. Evaluation of MAPK signaling pathway was done by Immuno histochemistry (IHC). Results Total 46 patients (Male=40 with mean age of 46.82 Y) were enrolled for the study with post- transplant duration of 33.72 months. The demographic details with mean values were: S Creatinine = 1.68 mg/dl, BUN = 28.26 mg/dl, Tac level = 5.98 ng/ml, S Uric Acid = 6.95 mg/dl, S Albumin = 3.46 g/dl, Na+/K+ = 128.9/4.16 mmol/L, S Phosphorus = 3.18 mg/dl, S Alkaline phosphate = 83.22 u/L. Induction regimen (Basiliximab=40, ATG= 6) Baseline Immunosuppression MMF+Pred (Tacrolimus/Cyclosporin) = 43/3. C4d were positive in 23 and DSA+ in 3 patients. Comparative miRnome analysis revealed that a total of 123 miRNAs showed differential level of expression in CABMR patients (n=3) as compared to the control (n=1) out of which 93 were upregulated while 40 were downregulated. Conserved level of differential expression shows upregulation of miR-9 (p≤0.0001), miR-21 (p=0.0024), miR-29a (p=0.0001), miR-126 (p=0.0076), miR-210 (p=0.0001) and miR-223 (p=0.0006) while hsa-miR-221 was downregulated. Pathway analysis shows involvement of MAPK signaling pathways, Transforming growth factor-β (TGF-β) superfamily signaling pathways, Wnt signaling pathways, ErbB signaling pathways. IHC of MAPK signaling shows significantly positive cytoplasmic tubular intensity as well as inflammatory cells intensity. Conclusion Altered miRNA expression profiling was found in CABMR compared to healthy controls. MAPK signaling pathways is involved in the pathogenesis of CABMR MicroRNAs are crucial regulators of cell function. They are easy to detect and represent potentially good targets for novel therapies.

Involvement of microRNA-124 in biological behaviors of laryngeal carcinoma via PLOD2 signaling pathway.

The purpose of this study was to investigate the role of microRNA-124 (miR-124) in laryngeal carcinoma (LC) and to explore the underlying mechanisms. LC tissues were collected from 98 patients diagnosed with LC in our hospital. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) analysis was used to detect the expression levels of miR-124 and procollagen-lysine, 2-oxoglutarate 5-dioxygenase 2 (PLOD2) in LC tissues and cell lines. Bioinformatics (TargetScan and miRDB) and double Luciferase assay were performed to predict and confirm the relationship between PLOD2 and miR-124 in LC, respectively. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay and the colony formation assay were designed to measure the proliferation ability of the cells. Additionally, transwell and wound healing assays were used to explore cell migration and invasion. MiR-124 was found significantly downregulated both in LC tissues and cells. PLOD2 was predicted and confirmed as the target gene of miR-124 in LC. By regulating the protein expression of PLOD2, miR-124 could significantly suppress the proliferation, migration, and invasion of cells. However, the transfection of PLOD2 could partially offset the effects of miR-124 on LC cells. MiR-124 was involved in regulating the malignant behaviors of LC cells. Furthermore, the miR-124/PLOD2 axis might become a potential target for the treatment of LC.

IL-6-miR-210 Suppresses Regulatory T Cell Function and Promotes Atrial Fibrosis by Targeting Foxp3.

The aim of this study was to explore the role of IL-6-miR-210 in the regulation of Tregs function and atrial fibrosis in atrial fibrillation (AF). The levels of interleukin (IL)-6 and IL-10 in AF patients were detected by using ELISA. Proportions of Treg cells were detected by fluorescence activated cell sorting analysis in AF patients. The expression of Foxp3, α-SMA, collagen I and collagen III were determined by western blot. The atrial mechanocytes were authenticated by vimentin immunostaining. The expression of miR-210 was performed by quantitative real-time polymerase chain reaction (qRT-PCR). TargetScan was used to predict potential targets of miR-210. The cardiomyocyte transverse sections in AF model group were observed by H&E staining. The myocardial filaments were observed by masson staining. The level of IL-6 was highly increased while the level of IL-10 (Tregs) was significantly decreased in AF patients as compared to normal control subjects, and IL-6 suppressed Tregs function and promoted the expression of α-SMA, collagen I and collagen III. Furthermore, miR-210 regulated Tregs function by targeting Foxp3, and IL-6 promoted expression of miR-210 via regulating hypoxia inducible factor-1α (HIF-1α). IL-6-miR-210 suppresses regulatory T cell function and promotes atrial fibrosis by targeting Foxp3.

Scanner invariant representations for diffusion MRI harmonization.

PurposeIn the present work, we describe the correction of diffusion‐weighted MRI for site and scanner biases using a novel method based on invariant representation.Theory and MethodsPooled imaging data from multiple sources are subject to variation between the sources. Correcting for these biases has become very important as imaging studies increase in size and multi‐site cases become more common. We propose learning an intermediate representation invariant to site/protocol variables, a technique adapted from information theory‐based algorithmic fairness; by leveraging the data processing inequality, such a representation can then be used to create an image reconstruction that is uninformative of its original source, yet still faithful to underlying structures. To implement this, we use a deep learning method based on variational auto‐encoders (VAE) to construct scanner invariant encodings of the imaging data.ResultsTo evaluate our method, we use training data from the 2018 MICCAI Computational Diffusion MRI (CDMRI) Challenge Harmonization dataset. Our proposed method shows improvements on independent test data relative to a recently published baseline method on each subtask, mapping data from three different scanning contexts to and from one separate target scanning context.ConclusionsAs imaging studies continue to grow, the use of pooled multi‐site imaging will similarly increase. Invariant representation presents a strong candidate for the harmonization of these data.

MicroRNA-489 targets XIAP to inhibit the biological progression of ovarian cancer via regulating PI3K/Akt signaling pathway and epithelial-to-mesenchymal transition.

Ovarian cancer (OC) is a deathful malignant tumor in women worldwide, and its poor prognosis mainly results from metastasis. Recently, microRNA (miRNA/miR) has been found to exert crucial functions in the progression of multiple tumors by affecting expressions of their targets. However, the biological roles and the potential mechanism of miR-489 in OC need further elucidation. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was utilized to confirm the miR-489 expressions in OC tissue samples and cell lines. The functions of miR-489 were analyzed by performing functional assays, such as MTT (3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assays and transwell assays. The downstream target of miR-489 was confirmed by TargetScan and luciferase reporter assay. Western blot was conducted to detect the expression of indicators associated with the down-stream signaling pathway. MiR-489 was prominently downregulated in OC tissues and cells, and the decreased miR-489 expression was related to malignant clinicopathologic features and poor prognosis of OC patients. Functional assays demonstrated that miR-489 could suppress OC cell viability, invasion, and migration. X-linked inhibitor of apoptosis protein (XIAP) was identified as a target of miR-489 and partially regulated the functions of miR-489 in OC. Moreover, we found that miR-489 inhibits OC progression via regulating phosphatidyl-inositol 3-kinase/protein kinase B pathway (PI3K/AKT) and epithelial-to-mesenchymal transition (EMT). Our results demonstrated that miR-489 inhibited OC development by directly binding to XIAP and regulating PI3K/Akt and EMT signal pathways, and miR-489 might serve as a promising biomarker for OC treatment in the future.

MiR-630 Regulates Proliferation and Apoptosis of Clear Cell Renal Cell Carcinoma Through Targeted Binding to Phosphatase and Tensin Homolog

Clear cell renal cell carcinoma (ccRCC) occurs as tumor cells in the kidney quickly accumulate and forms a lump (mass). Previous evidence showed that miR-630 overexpression is associated with gastric cancer progression. In this study, the differentially expressed miRNAs were screened in ccRCC patients using the gene expression profile chip. The possible targets for miR-630 were predicted by Target Scan and assessed via the luciferase reporter gene assay. The cell proliferation and apoptosis were detected via CCK-8 assay and flow cytometry. The Bcl-2, Bax, PTEN as well as downstream PI3K/AKT/mTOR signaling were detected via Western blotting. The result revealed that in silico analysis using the Target Scan software predicted that PTEN might be a potential target for miR-630, which was further verified via the luciferase reporter gene assay. The increase of miR-630 expression significantly raised the proliferation rate (P &lt; 0.05) and reduced cellapoptosis (P &lt; 0.05). In addition, the results of Western blotting also revealed that miR-630 mimic could activate PI3K/AKT/mTOR signaling (P &lt; 0.05). In conclusion, miR-630 cantargeted reduce the PTEN expression, and then activate the PI3K/AKT/mTOR signaling, ultimately promoting cell proliferation and inhibiting apoptosis of ccRCC.

Bioinformatic analysis of peripheral blood miRNA of breast cancer patients in relation with anthracycline cardiotoxicity

BackgroundThe current diagnostic methods and treatments still fail to lower the incidence of anthracycline-induced cardiotoxicity effectively. In this study, we aimed to (1) analyze the cardiotoxicity-related genes after breast cancer chemotherapy in gene expression database and (2) carry out bioinformatic analysis to identify cardiotoxicity-related abnormal expressions, the biomarkers of such abnormal expressions, and the key regulatory pathways after breast cancer chemotherapy.MethodsCardiotoxicity-related gene expression data (GSE40447) after breast cancer chemotherapy was acquired from the Gene Expression Omnibus (GEO) database. The biomarker expression data of women with chemotherapy-induced cardiotoxicity (group A), chemotherapy history but no cardiotoxicity (group B), and confirmatory diagnosis of breast cancer but normal ejection fraction before chemotherapy (group C) were analyzed to obtain the mRNA with differential expressions and predict the micro RNAs (miRNAs) regulating the differential expressions. The miRanda formula and functional enrichment analysis were used to screen abnormal miRNAs. Then, the Gene Ontology (GO) analysis was adapted to further screen the miRNAs related to cardiotoxicity after breast cancer chemotherapy.ResultThe data of differential analysis of biomarker expression of groups A, B, and C using the GSE40447-related gene expression profile database showed that there were 30 intersection genes. The differentially expressed mRNAs were predicted using the miRanda and Target Scan software, and a total of 2978 miRNAs were obtained by taking the intersections. Further, the GO analysis and targeted regulatory relationship between miRNA and target genes were used to establish miRNA-gene interaction network to screen and obtain seven cardiotoxicity-related miRNAs with relatively high centrality, including hsa-miR-4638-3p, hsa-miR-5096, hsa-miR-4763-5p, hsa-miR-1273 g-3p, hsa-miR6192, hsa-miR-4726-5p and hsa-miR-1273a. Among them, hsa-miR-4638-3p and hsa-miR-1273 g-3p had the highest centrality. The PCR verification results were consistent with those of the chip data. There are differentially expressed miRNAs in the peripheral blood of breast cancer patients with anthracycline cardiotoxicity. Among them, hsa-miR-4638-3p and hsa-miR-1273 g-3p are closely associated with the onset of anthracycline cardiotoxicity in patients with breast cancer. The signaling pathway is mainly concentrated in TGF-β signaling pathway and adhesion signaling pathway.ConclusionsChanges in expression of hsa-miR-4638-3p and hsa-miR-1273 g-3p may contribute to the detection of anthracyclines induced cardiac toxicity, and their potential function may be related to TGF-β signaling pathway and adhesion signaling pathway.

MiR-155-5p accelerates cerebral ischemia-reperfusion injury via targeting DUSP14 by regulating NF-κB and MAPKs signaling pathways.

This study aimed to explore the role of miR-155-5p in middle cerebral artery occlusion/reperfusion (MCAO/R) model in rats and oxygen-glucose deprivation/reoxygenation (OGD/R)-induced SH-SY5Y cells. In addition, this study also aimed to explore the underlying mechanisms to expect that miR-155-5p may be investigated as a new and effective diagnostic and therapeutic target for ischemic stroke. The in vivo MCAO/R rat model and in vitro OGD/R cell model were established. The miR-155-5p mRNA expression was detected by quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Dual specificity ATPase (DUSP) 14 was predicted to be a potential target of miR-155-5p by TargetScan. The targeting relationship was confirmed by Luciferase assay. The cell viability was determined using the Cell Counting Kit-8 (CCK-8). The expression level of inflammatory cytokines, including tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) levels were detected by Enzyme-Linked Immunosorbent Assay (ELISA). Western blot was used to detect the protein expression of DUSP14, the apoptotic protein Cleaved cysteine-aspartic acid protease (caspase)-3, and Cleaved PARP, as well as nuclear factor kappa B (NF-κB) and MAPKs signaling pathways related proteins. MiR-155-5p was upregulated in both MCAO/R rats and OGD/R-induced SH-SY5Y cells. MiR-155-5p knockdown inhibited OGD/R-induced cell injury and inflammation, as well as MCAO/R-induced brain injury. MiR-155-5p regulated the NF-κB and MAPKs signaling pathways by targeting DUSP14. DUSP14 knockdown partially reversed the protective effect of miR-155-5p knockdown on OGD/R-induced SH-SY5Y cell injury and inflammation. MiR-155-5p accelerates cerebral I/R injury via targeting DUSP14 by regulating NF-κB and MAPKs signaling pathways. Inhibition of miR-155-5p significantly reduces apoptosis and brain injury. These results indicated that miR-155-5p plays a key role in cerebral I/R injury and has the potential to be explored as a new target for ischemic stroke.

Long non-coding RNA XIST protects chondrocytes ATDC5 and CHON-001 from IL-1β-induced injury via regulating miR-653-5p/SIRT1 axis.

Chondrocyte apoptosis is linked to cartilage degeneration, and considered as a crucial event during the development of osteoarthritis (OA). X inactive specific transcript (XIST) is an oncogenic long non-coding RNA (lncRNA). However, its role in the pathophysiological process of OA remains largely unknown. In this work, quantitative real-time reverse transcriptase PCR (qRT-PCR) was employed to measure the expression of XIST, miR-653-5p and sirtuin1 (SIRT1) mRNA in OA and normal cartilage tissues. Chondrocyte cell lines, CHON-001 and ATDC5, were treated with different doses of interleukin- 1β (IL-1β) to mimic the inflammatory environment of OA in vitro. Overexpression plasmids, microRNA (miRNA) mimics, miRNA inhibitors and small interfering RNAs (siRNAs) were constructed and transfected into CHON-001 and ATDC5 cells. 3-(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay was adopted to determine the cell viability. Western blot was used to detect the expression of apoptosis-related proteins. Enzyme-linked immunosorbent assay (ELISA) was employed to probe the expression levels of inflammatory factors. Flow cytometry was used to analyze the cell apoptosis. StarBase and TargetScan databases were used to predict the binding sites between XIST and miR-653-5p, miR-653-5p and 3'UTR of SIRT1, respectively, which were then verified by dual luciferase reporter assay. The data in the present study demonstrated that XIST and SIRT1 were down-regulated while miR-653-5p was up-regulated in OA tissues and cell models. The up-regulation of XIST increased the viability of CHON-001 and ATDC5 cells, while it impeded their apoptosis and inflammatory response induced by IL-1β. Conversely, miR-653-5p had opposite effects. It was proved that miR-653-5p could be sponged and suppressed by XIST. Additionally, SIRT1 was identified as a target of miR-653-5p, and SIRT1 could be suppressed by XIST indirectly. In conclusion, down-regulated XIST was involved in the injury of chondrocytes during the pathophysiological process of OA, and XIST up-regulation protected chondrocytes from inflammatory injury via regulating miR-653-5p/SIRT1 axis.

LncRNA LINC01278 accelerates colorectal cancer progression via miR-134-5p/KDM2A axis.

Long non-coding RNAs (lncRNAs) play vital roles in the pathogenesis and development of multiple cancers, including colorectal cancer (CRC). Nevertheless, the regulatory mechanisms of LINC01278 in CRC remain unknown. Our research aims to identify the regulatory mechanisms of LINC01278 in CRC. The expression of LINC01278 was examined by quantitative real-time polymerase chain reaction (RT-qPCR). StarBase and TargetScan websites were used to predict the interaction between miR-134 and LINC01278 or KDM2A, which was further confirmed by Dual-Luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Cell viability, migration, and invasion were detected by Cell Counting Kit-8 (CCK-8) and transwell assays. LINC01278 was upregulated in CRC tissues and cell lines, and knockdown of LINC01278 suppressed CRC cell progression. In addition, LINC01278 inhibited miR-134 expression by direct interaction, and the inhibition of miR-134 abolished the suppressive effects of LINC01278 knockdown on viability, migration, and invasion of CRC cells. Furthermore, KDM2A was confirmed to be a target gene of miR-134. Overexpression of KDM2A facilitated the tumorigenesis of CRC, while this effect was reversed by the upregulation of miR-143. Finally, it was demonstrated that miR-134 inhibitor reversed the shLINC01278‑mediated inhibitory effect on KDM2A expression. Our study demonstrated that LINC01278 upregulated KDM2A to promote CRC progression by interacting with miR-143, suggesting that LINC01278 might be a new therapeutic target of CRC.

MiR-188 inhibits proliferation and promotes apoptosis of lung adenocarcinoma cells by targeting SIX1 to negatively regulate ERK signaling pathway.

To explore the effects of micro ribonucleic acid (miR)-188 on proliferation and apoptosis of lung adenocarcinoma (LUAD) cells, and its potential mechanism. The expression level of miR-188 in LUAD cell lines was detected via quantitative Real Time-Polymerase Chain Reaction (PCR). The effects of miR-188 overexpression on proliferation and apoptosis of A549 cells were detected using methyl thiazolyl tetrazolium (MTT) assay, colony formation assay, and flow cytometry. The potential targets for miR-188 were predicted using the TargetScan Human database, and the interaction between miR-188 and target gene was determined through Dual-Luciferase reporter assay. Moreover, the associations of miR-188 and sine oculis homeobox homolog 1 (SIX1) with the extracellular signal-regulated kinase (ERK) pathway were detected via Western blotting. The expression of miR-188 significantly declined in LUAD cell lines (p<0.05). The overexpression of miR-188 significantly reduced the proliferation rate of A549 cells and increased the percentage of apoptotic A549 cells (p<0.05). Similarly, it was found in colony formation assay that the overexpression of miR-188 inhibited the colony formation ability of A549 cells most significantly (p<0.05). SIX1 was a direct target for miR-188, and its mRNA and protein expressions were downregulated by the overexpression of miR-188. The remarkable downregulation of phosphorylated ERK was observed in A549 cells overexpressing miR-188, while the decline in phosphorylated ERK was reversed in A549 cells overexpressing miR-188 and SIX1. The expression of miR-188 is downregulated in LUAD cell lines. The overexpression of miR-188 inhibits proliferation and promotes apoptosis of LUAD cells, whose functional mechanism may be related to its regulation on the ERK signaling pathway by targeting SIX1.

MiR-5195-3p inhibits the proliferation of glioma cells by targeting BIRC2.

The aim of this study was to investigate the effects of microRNA-5195-3p (miR-5195-3p) on the proliferation of glioma cells and to explore its related mechanisms. The expression level of miR-5195-3p in glioma tissues and cell lines was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Online prediction websites (TargetScan and miRanda) were used to screen the potential targets of miR-5195-3p. Luciferase reporter gene assay and Western blot were performed to confirm the targets of miR-5195-3p. Furthermore, the effects of miR-5195-3p on cell proliferation were detected by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide), colony formation assay, and flow cytometry, respectively. MiR-5195-3p was lowly expressed in both glioma tissues and cells. Baculoviral IAP repeat-containing 2 (BIRC2) was identified as a direct target of miR-5195-3p. Over-expression of miR-5195-3p in glioma cells significantly decreased the protein expression of BIRC2. Besides, the proliferative capacity and colony formation ability were significantly inhibited after transfection of miR-5195-3p in vitro. In addition, flow cytometry indicated that an evident G1 phase arrest occurred in miR-5195-3p over-expressed group. In this work, we emphasized the suppressor function of miR-5195-3p in the proliferation of glioma cells. Furthermore, our findings provided an experimental basis for the research and treatment of glioma.