Expression Levels Of miR-146a Research Articles

MicroRNA-181a mediates the chemo-sensitivity of glioblastoma to carmustine and regulates cell proliferation, migration, and apoptosis

Glioblastoma multiform (GBM) as the most frequent and lethal brain tumor is defined by aggressive invasiveness and considerable resistance to chemotherapy. The molecular mechanisms underlying GBM tumorigenesis still needs to be further investigated. Considering that, the current study was aimed to investigate the function of miR-181a in human glioblastoma cells in combination with carmustine. U373 cell line with the low expression levels of miR-181a was selected for functional investigations. MTT assay was used to determine cell viability and Annexin V/PI and DAPI staining were employed to evaluate apoptosis induction. Also, cell migration and cell cycle progression were investigated using wound healing test and flow cytometry, respectively. qRT-PCR was used for the quantification of gene expression. MTT assay results revealed that miR-181a replacement increased the sensitivity of U373 cells to low doses of carmustine. Moreover, miR-181a was shown to increase the sub G1 cell cycle arrest and apoptosis induction by carmustine via regulating the expression of related genes including caspase-9, Bcl-2, and SIRT1. Furthermore, this miRNA combined with carmustine suppressed cell migration via downregulation of MMP-2 and Bach1 and reduced the clonogenic ability of U373 cells. Additionally, miR-181a-mediated downregulation of AKT1 implied that this miRNA could inhibit cell proliferation by modulating PI3K/AKT signaling pathway. In conclusion, the findings of this study suggest that miR-181a replacement, regarding its tumor-suppressive effects and sensitization of glioblastoma cells to carmustine, could be considered as a potential therapeutic strategy to improve the efficiency of glioblastoma chemotherapy.

Potential role of MicroRNA as a diagnostic tool in the detection of bovine mastitis

Bovine mastitis is a major health problem that affects dairy cows and has a negative impact on milk production. The presence of microRNAs in biofluids, such as blood and milk, could play a pivotal role in the detection of bovine mastitis. The purpose of the current study was to determine the levels of microRNA gene expression in milk, in combination with other reported mastitis indicators, as a biomarker of bovine mastitis. Milk samples (n = 171) were obtained from 113 dairy cows with known disease status (i.e., healthy; n = 23 cows, subclinical mastitis; n = 45 cows, or clinical mastitis; n = 45 cows) and analyzed for the presence of MIR24-2, MIR29B-2, MIR146A, MIR148A, MIR155, MIR181A1, MIR184, and MIR223 expression using the real-time PCR (qPCR) method. The expression data were then utilized in the creation of receiver operator characteristic curves (ROC) and further analyzed by the machine learning (ML) methods. MIR29B-2, MIR146A, MIR148A, and MIR155 expression levels differed significantly among the three groups. These potential microRNA biomarkers of mastitis exhibited high sensitivity and specificity. Next, we applied ML algorithm, specifically, a decision tree (DT) model to predict the status of milk based on MIR29B-2 and MIR146A expression levels. The results suggested that MIR29B-2, when used in combination with the California mastitis test (CMT) and days in milk (DIM) data, was applicable for screening and classification of milk samples from cows as healthy, subclinical mastitis, or mastitis. MIR29B-2 appears to have sufficient discriminatory power to enable it to be utilized as a biomarker in cases where the status of a milk sample cannot be determined based on CMT results.

Expression Level of MiR-146a in Acute Myeloid Leukemia Patients and Its Clinical Significance

To detect the expression of miR-146a in patients with AML, and to evaluate the relationship between miR-146a expression level and clinical characteristics, treatment response, EFS and OS. 154 patients with newly diagnosed AML were enrolled in AML group, 50 controls (patients with thrombocytopenic purpura or voluntary donor of bone marrow) were enrolled in control group. The miR-146a expression levels in bone marrow mononuclear cells was detected by RT-PCR between 2 group. AML patients were treated with chemotherapeutic drugs, and their clinical response and survivals were assessed. The expression level of MiR-146a in AML group was significantly lower than that in control group. The ROC showed that miR-146a could distinguish the patients in AML and control group better (area under curve 0.819 (95%CI: 0.761-0.877). Meanwhile, the proportion of good and moderate good prognosis (P＜0.001), proportion of WBC count ≤15.2×109/L (P＜0.05), CR rate (P＜0.05), EFS (P＜0.01) and OS (P＜0.01) in patients with high miR-146a expression were higher than those in patients with low miR-146a expression. Cox's model showed that miR-146a expression level positively realated with incressed EFS and OS. MiR-146a is downregulated in AML patients, which might be served as a biomarker for predicting risk and prognosis of AML patients.

MicroRNA Profiling in Wilms Tumor: Identification of Potential Biomarkers

Wilms tumor (WT) is the most frequently diagnosed malignant renal tumor in children. With current treatments, ~90% of children diagnosed with WT survive and generally present with tumors characterized by favorable histology (FHWT), whereas prognosis is poor for the remaining 10% of cases where the tumors are characterized by cellular diffuse anaplasia (DAWT). Relatively few studies have investigated microRNA-related epigenetic regulation and its relationship with altered gene expression in WT. Here, we aim to identify microRNAs differentially expressed in WT and describe their expression in terms of cellular anaplasia, metastasis, and association with the main genetic alterations in WT to identify potential prognostic biomarkers. Expression profiling using TaqMan low-density array was performed in a discovery cohort consisting of four DAWT and eight FHWT samples. Relative quantification resulted in the identification of 109 (48.7%) microRNAs differentially expressed in both WT types. Of these, miR-10a-5p, miR-29a-3p, miR-181a-5p, miR-200b-3p, and miR-218-5p were selected and tested by RT-qPCR on a validation cohort of 53 patient samples. MiR-29a and miR-218 showed significant differences in FHWT with low (P = 0.0018) and high (P = 0.0131) expression, respectively. To discriminate between miRNA expression FHWTs and healthy controls, the receiver operating characteristic (ROC) curves were obtained; miR-29a AUC was 0.7843. Furthermore, low expression levels of miR-29a and miR-200b (P = 0.0027 and P = 0.0248) were observed in metastatic tumors. ROC curves for miR-29a discriminated metastatic patients (AUC = 0.8529) and miR-200b (AUC = 0.7757). To confirm the differences between cases with poor prognosis, we performed in situ hybridization for three microRNAs in five DAWT and 17 FHWT samples, and only significant differences between adjacent tissues and FHWT tumors were found for miR-181a, miR-200b, and miR-218, in both total pixels and nuclear analyses. Analysis of copy number variation in genes showed that the most prevalent alterations were WTX (47%), IGF2 (21%), 1q (36%) gain, 1p36 (16%), and WTX deletion/1q duplicate (26%). The five microRNAs evaluated are involved in the Hippo signaling pathway and participate in Wilms tumor development through their effects on differentiation, proliferation, angiogenesis, and metastasis.

MicroRNA-146a Is a Wide-Reaching Neuroinflammatory Regulator and Potential Treatment Target in Neurological Diseases.

Progressive functional deterioration and loss of neurons underlies neurological diseases and constitutes an important cause of disability and death worldwide. The causes of various types of neurological diseases often share several critical nerve-related cellular mechanisms and pathological features, particularly the neuroinflammatory response in the nervous system. A rapidly growing body of evidence indicates that various microRNAs play pivotal roles in these processes in neurological diseases and might be viable therapeutic targets. Among these microRNAs, microRNA-146a (miR-146a) stands out due to the rapid increase in recent literature on its mechanistic involvement in neurological diseases. In this review, we summarize and highlight the critical role of miR-146a in neurological diseases. MiR-146a polymorphisms are associated with the risk of neurological disease. Alterations in miR-146a expression levels are crucial events in the pathogenesis of numerous neurological diseases that are spatially and temporally diverse. Additionally, the target genes of miR-146a are involved in the regulation of pathophysiological processes in neurological diseases, particularly the neuroinflammatory response. In summary, miR-146a mainly plays a critical role in neuroinflammation during the progression of neurological diseases and might be a prospective biomarker and therapeutic target. Understanding the mechanisms by which miR-146a affects the neuroinflammatory response in different neurological injuries, different cell types, and even different stages of certain neurological diseases will pave the way for its use as a therapeutic target in neurodegenerative diseases.

Effects of dexmedetomidine pretreatment on rats with sepsis-induced acute kidney injury and miR-146a expression

The current research aimed to study the effects of dexmedetomidine (DEX) pretreatment on rats with sepsis-induced acute kidney injury (SAKI) and miR-146a expression. The model of SAKI was established through the tail vein injection of lipopolysaccharide (LSP). We used an automatic biochemical analyzer to detect serum urea nitrogen (BUN) and creatinine (Cre) levels. The expression levels of urine KIM-1 and NGAL and serum IL-1β and IL-6 were analyzed by enzyme-linked immunosorbent assay (ELISA). The content and activity of superoxide dismutase (SOD) were detected by the xanthine oxidase method. The content of malondialdehyde (MDA) was determined by the thiobarbituric acid (TBA) method. Reactive oxygen species (ROS) was detected by fluorescent probe DCFH-DA. Catalase (CAT) was detected by potassium permanganate titration. The expression level of miR-146a in the renal tissue and serum was detected by RT-PCR. The expression levels of Nrf2 and HO-1 proteins were detected by Western blot. Compared with those in the model group, rats in the DEX group had significantly lower expression levels of serum BUN, Cre, IL-1β, and IL-6, and oxidant markers MDA and ROS, but significantly higher expression levels of miR-146a and antioxidant markers SOD and CAT. DEX pretreatment could improve the kidney morphology, injury severity, and Nrf2 and HO-1 proteins of rats with SAKI. In conclusion, DEX can improve oxidative stress and inflammatory responses in rats with SAKI, reduce the severity of the renal injury, and up-regulate the expression level of miR-146a.

SUN-LB127 In Experimental Peripheral Arterial Disease MIR29a Modulation Improves Perfusion Recovery and Muscle Function to a Greater Extent Than ADAM12 Augmentation in Mice With Type 2 Diabetes

Diabetes Mellitus (DM) is a major risk factor for developing peripheral arterial disease (PAD) and individuals with DM have worse PAD outcomes but the molecular mechanisms involved are poorly understood. We previously identified a disintegrin and metalloproteinase gene 12 (ADAM12) as a key genetic modifier of post-ischemic perfusion recovery in experimental PAD that it is regulated by miR29a through direct interaction. In ischemic hind limbs non-diabetic mice, miR29a expression is downregulated and allows ADAM12 upregulation. However, in ischemic hind limbs of type 2 DM (DM2) mice miR29a expression remains elevated and prevents ADAM12 upregulation. Hence, we hypothesized that inhibition of miR29a or augmenting ADAM12 would improve the impaired perfusion recovery typical of mice with DM2 in experimental PAD. Here, in mice with DM2 we compare the therapeutic effectiveness of miR29a inhibition (miR29aINH) to ADAM12 gene overexpression on perfusion recovery in experimental PAD.26 - 28 weeks old C57BL/6 mice on high fat diet were randomized into 3 treatment groups: sham (saline), miR29aINH, or ADAM12 plasmid. Agents were delivered by micro-bubbles at 5 days and 1 day pre-surgery. Experimental PAD was achieved by unilateral ligation and excision of the femoral artery of the left hind limb. The right hind limb serves as non ischemic control. Peripheral blood flow is measured by Laser Speckle Contrast Imaging. Tibialis anterior contractile function was assessed using an Aurora 300B-LR. ADAM12 and miR29a expression levels were quantified using qPCR on day 3 post surgery since perfusion at this point is similar between the experimental groups. Data are expressed as means ± SEM. Statistical significance of P < 0.05 as significant is performed with the independent Student’s t-test.Gene expression analysis 3 days post ischemia showed miR29a inhibitor reduced miR29a levels (miR29aINH: 0.70±0.06, n=5, ADAM12: 0.62±0.06, n=4, p=0.3938 and Sham: 1.02±0.06, n=5, p= 0.004 for miR29aINH vs Sham) and increased ADAM12 expression to a greater extent than ADAM12 gene transfer (miR29aINH: 208.62±24.52, n=3, ADAM12: 20.71±2.98, n=4, p= 0.0003 and Sham: 11.75±4.94, n=4). 3 weeks post-surgery, there was significantly higher perfusion recovery in mice treated with miR29aINH compared to ADAM12 gene transfer (miR29aINH: 97.59±6.14, n= 10, ADAM12: 58.45±4.87, n=8, p=0.002 and Sham: 42.52±5.35, n=5). Muscle contraction in the ischemic limb was >2-fold stronger in the miR29aINH treated mice compared to ADAM12 gene transfer (miR29aINH: 0.54±0.08, n=7, ADAM12: 0.26±0.06, n=7, p= 0.0170 and Sham: 0.17±0.06, n=6).Our result reveals for the first time that inhibiting elevated microRNAs may be a more effective therapeutic approach than gene transfer to improve blood flow and muscle function in type 2 DM and PAD.

Clinical role of MiRNA 29a and MiRNA 335 on breast cancer management: their relevance to MMP2 protein level

Background Circulating miRNAs are novel biomarkers, authors aimed to investigate the expression level of miR-29a and miR-335 and their relevance to CEA, CA15.3, and matrix metalloproteinase-2 (MMP2). Materials and methods Breast cancer (BC) patients (n = 44), benign breast lesion patients (n = 25), and healthy individuals (n = 19) were enrolled for detection of miRNA expression levels, MMP2 and biochemical markers using quantitative polymerase chain reaction (PCR) and ELISA, respectively. Results Expression of miR-29a and miR-335 were significantly decreased in breast patients as compared to healthy individuals, while biochemical markers were high in BC patients as compared to the other two groups. The diagnostic efficacy for miR-29a, miR-335, and MMP2 were superior to both CEA and CA 15.3 for early detection of BC patients. Conclusions Detection of the miR-29a and miR335 expression levels in serum samples are significant promising biomarkers for BC diagnosis.

Expression and Clinical Significance of MiR-146a and MiR-221 in Childhood Acute T Lymphoblastic Leukemia

To investigate the expression of miR-146a and miR-211 in peripheral blood mononuclear cells (PBMNC) of children with acute T-lymphoblastic leukemia (T-ALL) and their clinical significance. One hundred and thirty newly diagnosed children with T-ALL from Hainan Third People's Hospital (T-ALL group) and 50 healthy persons (control group) were selected. The newly diagnosed T-ALL patients before treatment were taken as initial group, the patients reachived complete remission after induction therapy for 33 days were taken as remission group (n=98), the patients not reachived complete remission after induction therapy fore 33 days were taken as rafractory group (n=32). The expression levels of miR-146a and miR-221 in PBMNC were detected by real-time fluorescence quantitative PCR. ROC curve analysis was used to evaluate the diagnostic value of miR-146a and miR-221 for T-ALL, Pearson correlation analysis was used to estimate the correlation of miR-146a and miR-221 expression with white blood cell count in children with T-ALL. The expression levels of miR-146a and miR-221 of PBMNC in T-ALL group were significantly higher than those in control group (5.83±1.54 vs 0.96±0.17) (7.13±2.60 vs 1.64±0.51) (P＜0.01). The expression levels of miR-146a and miR-221 in refractory group were significantly higher than those in remission group and initial group (8.74±2.35 vs 1.70±0.63 and 5.83±1.54) (11.316±4.83 vs 2.62±0.85 and 7.13±2.60) (P＜0.01). The expression levels of miR-146a and miR-221 correlated with white blood cell count, risk typing and MRD (P＜0.05). ROC curve analysis showed that the best cutoff values of miR-146a and miR-221 for diagnosing childhood T-ALL were 3.90 and 5.28, resoectively. While the AUC (95% CI) of the T-ALL jointy diagnosed by miR-146a and miR-221 was 0.835 (0.764-0.892), and it's sensitivity and specificity were 85.0% and 77.2%, respectively. The correlation analysis showed that the expression levels of miR-146a and miR-221 in PBMNC of children with T-ALL positively correlated with the white blood cell count (r=0.705, r=0.653, P＜0.01), and that of miR-146a positively correlated with miR-221 (r=0.784, P＜0.01). The expression of miR-146a and miR-221 is up-regulated in children with T-ALL and closely relates with the prognosis of children with T-ALL. The combined detection of miR-146a and miR-221 is certain value for diagnosis of T-ALL.

Expressions and clinical significances of miR-15 and miR-29a in serum of patients with diabetic retinopathy

Objective To analyze the expression levels of miR-15 and miR-29a in serum of patients with diabetic retinopathy (DR) and their diagnostic values for DR. Methods 155 patients (155 eyes) with type 2 diabetes mellitus (DM) were treated in our hospital from June 2016 to August 2018, according to the occurrence of retinopathy and the degree of retinopathy, the patients were divided into five groups: 50 cases of non-retinopathy group (NDR group), 56 cases of simple retinopathy group (SDR group), 49 cases of proliferative retinopathy group (PDR group) and another 50 healthy persons in the same period were selected as the control group. Triglyceride (TG), total cholesterol (TC), and fasting blood glucose (FPG) were measured. Levels of serum miR-15 and miR-29a were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Pearson correlation method was used to analyze the relationships between levels of serum miR-15 and miR-29a with clinical indicators in DR patients. Receiver operating characteristic curve (ROC) was used to evaluate the diagnostic values of miR-15 and miR-29a in DR. Results The relative expression of miR-15 in serum of NDR group, SDR group, and PDR group were lower than that of control group (P<0.05), and the relative expression of miR-15 in NDR group, SDR group, and PDR group decreased gradually (P<0.05); the relative expression of miR-29a in serum of NDR group, SDR group, and PDR group were higher than that of control group (P<0.05), and the mRNA relative expression of miR-29a in NDR group, SDR group, and PDR group increased gradually (P<0.05); the expression of serum miR-15 in DR patients was negatively correlated with urinary albumin/creatinine (UACR), glycosylated hemoglobin (HbA1c) and the course of DM (r=-0.732, -0.492, -0.589, P<0.05); the expression level of miR-29a was positively correlated with UACR, HbA1c and the course of DM (r=0.744, 0.508, 0.556, P<0.05); the areas under the ROC curve (AUC) of miR-15 and miR-29a for DR diagnosis were 0.796 (95% CI: 0.724-0.857) and 0.677 (95% CI: 0.597-0.749), with diagnostic thresholds 0.63 and 1.11, sensitivities 84.9% and 53.8%, specificities 65.3% and 79.6%, repectively; miR-29a was a risk factor for DR, while miR-15 was a protective factor for DR. Conclusions The expression of mir-15 and miR-29a in the serum of DR patients is decreased, which is related to the degree of retinopathy and can be used as biomarkers for early diagnosis of DR. Key words: Diabetic retinopathy; miR-15; miR-29a; Early diagnosis

Evaluation of Micro-RNA Levels, Apoptosis and Oxidative Stress Markers in Patients Recieving Chemotherapy.

The primary objective of this study was to compare oxidative DNA damage markers, apoptosis markers and changes in miRNA levels in patients diagnosed with cancer and treated through chemotherapy. Our secondary objective was also to evaluate tumor responses that can be determined after post-chemotherapy clinical evaluations by physical examinations, laboratory results and radiological imagings, and to compare the clinical results to oxidative stress and apoptosis markers and micro RNA levels. To do that we designed a prospective observational cross-sectional study. A total of 34 cancer patients and 27 healthy controls were included in the study from the Harran University School of Medicine Department of Oncology. Newly diagnosed chemotherapy or radiotherapy naive patients without any chronic diseases were included into the study. Patients with a poor performance status (ECOG 2 and 3) and patients who did not meet the inclusion criteria were excluded. The cancer patients received chemotherapy according to their scheduled periods. Blood samples were taken from the patients before the first chemotherapy course and before the second chemotherapy round. Patients were called for toxicity control on the 10th day after the chemotherapy. Pre-chemotherapy, post-chemotherapy and control group miR-29a expression levels, change in apoptosis markers and oxidative DNA damage markers were obtained and compared. We studied 8-hydroxy 2-deoxyguanosine, total oxidant status, total anti-oxidant status, and oxidative status index for oxidative stress markers. We studied M30 and M65 as apoptosis markers. Clinical results of efficiency of the chemotherapy was acquired and compared to biochemical markers based on chemotherapy results. Chemotherapy toxicities were recorded. As a result, we found oxidative DNA damage markers and apoptosis markers were high in the cancer group, demonstrating that oxidative DNA damage and apoptosis might play a direct or indirect role in cancer etiology. However, there were subtle differences between pre-chemotherapy and post-chemotherapy levels. Mir-29a expressions were lower in cancer patients as compared to controls. However, the expression levels were not significantly change in pre- and postchemotherapy status. Moreover, we found no relationship between clinical status of patients (progression and regression) and studied biochemical markers. Thus, checking for DNA damage markers and taking precautions to lower the levels of these markers in individuals with cancer risk may be helpful in preventing cancer.

Chemical composition, antioxidant, antimicrobial and anti-inflammatory activity of Prunus spinosa L. fruit ethanol extract

Abstract Prunus spinosa L. (from Italy) fruit ethanol extract (40 µg/mL) was assessed by evaluating the antioxidant, anti-inflammatory and antimicrobial activities against five bacterial and two fungi ATCC strains. Moreover, the phenolic profile was also investigated and results are indicative of an intense anthocyanin accumulation which may be responsible for the antioxidant properties revealed by the DPPH assay. MIC and MBC/MFC values (4.36–8.72 mg/mL; 8.72–17.44 mg/mL, respectively) revealed a wide antibacterial activity and yeast inhibition. No specific inhibitory action was observed against the tested Gram-negative or Gram-positive bacteria. Preliminary data on the effect on both miR-126 and miR-146a expression levels suggested a very interesting anti-inflammatory activity of the extract. A possible mechanism underpinning the observed effects was hypothesized and discussed. Finally, P. spinosa fruit extract could be used as supplementary source of functional additives and might be a promising antimicrobial compound of natural origin to be employed to fight microbial resistance.

MicroRNA-146a attenuates the development of morphine analgesic tolerance in a rat model

ABSTRACT Background: Morphine plays an irreplaceable role in relieving severe pain clinically, while long-term medication inevitably leads to drug resistance. MicroRNA (miR) 146a has been reported to be a negative regulator in the process of morphine-tolerance formation. This study aimed to investigate how miR-146a affects the development of morphine analgesic tolerance. Methods: The morphine-tolerance rat model was established by means of one-week continuous morphine administration. Paw withdrawal latency test was performed every day, and spinal cord samples were dissected on the seventh day for Q-PCR and Western blotting to detect the expression level of miR-146a, and IRAK1/TRAF6 participated in TLR4 signaling pathway. Results: The expression of miR-146 was significantly decreased in morphine-tolerant model. Also, overexpression of miR-146a reduced the resistance caused by morphine, followed by the down-regulation of IRAK1/TRAF6 in TLR4 pathway. The inhibition of miR-146a remarkably decreased paw withdrawal latency as well as increased the expression levels of TLR4 signaling pathway-related molecules, IRAK1 and TRAF6. Conclusion: This study suggests that miR-146a attenuates morphine tolerance by inhibiting the expression of IRAK1/TRAF6 in TLR4 pathway, which could provide an essential experimental basis for the settlement of morphine resistance-associated matters.

The single nucleotide variant n.60G>C in the microRNA-146a associated with susceptibility to drug-resistant epilepsy

ObjectiveThe aim of this study was to evaluate single nucleotide variants (SNVs) n.-411A > G (rs57095329) and n.60 G > C (rs2910164) in microRNA (miR)-146a, related to suppressing of TRAF6 with risk for epilepsy, as well as miR-146a and TRAF6 levels. MethodsDNAs were extracted from epileptogenic tissues and blood leukocytes from drug-resistant epilepsy patients and healthy-individuals, respectively. Genotypes were identified by real-time PCR. Hardy-Weinberg equilibrium (HWE) and Fisher or X2 tests evaluated the difference between groups. The disease risk was assessed by odds ratio (OR) with 95 % confidence interval (95 %CI). The prognostic impact on probability seizure-free survival (PSF) was evaluated by Kaplan-Meier and log-rank tests. ResultsFor rs57095329 both control and patient samples were not in HWE (p < 0.05) and the genotypes prevalence was similar in patients and controls (p>0.05). For rs2910164, control samples were in HWE (p = 0.61), contrasting with patients (p = 0.03), and similar frequencies of wild-type homozygous (GG) (43.4 % vs. 34.4 %, p = 0.2) and variant (CC) genotypes (8.0 % vs. 6.6 %, p = 0.6) were observed in patients and controls, respectively. However, increased frequency of heterozygous (GC) was observed in patients compared to controls (59.0 % vs. 42.7 %, p = 0.04) with 1.98 (95 %CI=1.09-3.57) risk for epilepsy. The miR-146a expression level in the epileptogenic tissues was lower in the GC (p = 0.02) and CC (p = 0.09) compared to GG genotype. TRAF6 expression level was higher in CC than in GG genotype (p = 0.09). Interestingly, there was an increased frequency of patients harboring GC genotype and less time until surgery compared to patients harboring GG or CC (36.06 % vs. 11.5 %, p = 0.01), confirmed by PSF (p = 0.04). ConclusionsThe GC genotype for SNV rs2910164 appears associated with susceptibility to drug-resistant epilepsy due to the decreased MIR146a expression, favoring NF-kB pathway through TRAF6.

Overexpression of miR-146a inhibits the apoptosis of hippocampal neurons of rats with cerebral hemorrhage by regulating autophagy.

In this study, to investigate the effect of overexpression of miR-146a on autophagy of hippocampal neurons in rats with intracerebral hemorrhage (ICH), 72 Sprague-Dawley rats were randomly divided into the sham, ICH, miR-146a agomir, and miR-146a agomir control groups. The ICH model was constructed by injection of collagenase VII. The apoptosis of hippocampal neurons was measured by TUNEL assay. The levels of LC3 and Beclin 1 were analyzed by immunohistochemistry. Mitochondrial autophagy was examined by transmission electron microscopy. The levels of LC3A, LC3B, Beclin 1, Bax, Bcl-2, and cleaved caspase 3 were examined by Western blot. Western blot was also used to evaluate the expression of nuclear factor κB signaling pathway-related factors. To examine the effect of autophagy inhibitor (3-methyladenine (3-MA)) on miR-146a-regulated apoptotic protein expression, 30 rats were further divided into the sham, ICH, miR-146a agomir, 3-MA, and miR-146a + 3-MA groups. The levels of Bax, Bcl-2, and cleaved caspase 3 were examined by Western blot. Compared with the sham group, the nerve function scores, brain water content, the percentage of apoptotic cells, and the expression levels of LC3, Beclin 1, Bax, cleaved caspase 3, and p-P65 in the hippocampus of rats in the ICH group were all significantly increased (p < 0.05), whereas the expression levels of miR-146a, Bcl-2, and p-IκBα were markedly decreased (p < 0.05). Mitochondrial autophagy was also evident. Furthermore, compared with the ICH group, the results of the abovementioned tests in the miR-146a agomir group were reversed. The overexpression of miR-146a inhibited the autophagy of hippocampal neurons in rats with ICH.

Reduced miR-146a Promotes REG3A Expression and Macrophage Migration in Polymyositis and Dermatomyositis.

Background: Growing evidence from studies elsewhere have illustrated that microRNAs (miRNAs) play important roles in polymyositis and dermatomyositis (PM/DM). However, little has been reported on their relationship with regenerating islet-derived protein 3-alpha (REG3A) as well as their associative roles in macrophage migration. Therefore, this study sought to establish the association between miR-146a and REG3A as well as investigate their functional roles in macrophage migration and PM/DM pathogenesis.Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from PM/DM patients and healthy controls through density centrifugation. Macrophages were obtained from monocytes purified from PBMCs via differentiation before their transfection with miRNA or plasmids to investigate cell migration with transwell assay. An experimental autoimmune myositis murine model was used to investigate PM/DM. Real-time PCR and Western blot analysis were conducted to determine the expression levels of miR-146a, interferon gamma (IFN-γ), interleukin (IL)-17A, and REG3A.Results: The messenger RNA (mRNA) expression level of miR-146a markedly decreased, while the mRNA level of REG3A, IFN-γ, and IL-17A expression increased substantially in PBMCs from PM/DM patients compared with the healthy controls. The levels of IFN-γ and IL-17A in serum from PM/DM patients was much higher than the healthy controls. Immunohistochemistry analysis showed that REG3A expression increased in muscle tissues from patients. Consistent with clinical data, the mRNA expression level of miR-146a also decreased, whereas the mRNA and protein level of REG3A, IFN-γ, and IL-17A significantly increased in the muscle tissues of experimental autoimmune myositis mice. Moreover, miR-146a inhibited monocyte-derived macrophage migration, and REG3A promoted macrophage migration. In addition, IL-17A induced REG3A expression, while miR146a inhibited expression of REG3A in monocyte-derived macrophages from the PBMCs of the healthy donors. Notably, inhibition of macrophage migration by miR-146a was via the reduction in REG3A expression.Conclusions: Reduced miR-146a expression in PM/DM leads to increased REG3A expression that increases inflammatory macrophage migration, which may be a possible underlying mechanism of DM/PM pathogenesis.

MicroRNA-146a as a Prognostic Biomarker for Esophageal Squamous Cell Carcinoma.

Background and AimsMicroRNAs including miR146a have a regulatory role on the expression of genes and act with binding to 3ʹ-UTR region of the genes. Cyclooxygenase-2 (COX-2) is involved in carcinogenesis as an inflammatory marker, and microRNA-146a (miR-146a) as a negative regulatory factor. We aimed to evaluate miR146a expression as a prognostic or diagnostic biomarker for esophageal squamous cell carcinoma (ESCC) and also an association between miR146a and COX2 expression.Materials and MethodsWe quantified the level of miR-146a and COX-2 expression in cancerous and adjacent normal tissue samples obtained from 34 patients with ESCC, using real-time–PCR. Statistical analyses were conducted using one-sample t-test. Receiver-operating characteristic (ROC) curve and Kaplan–Meier analysis were applied to assay miR146a as a diagnostic and prognostic marker, respectively, during 4 years of the study. Furthermore, the Cox regression model was performed to assay the hazard ratio (HR). The association between miR-146a and COX2 expression level in ESCC patients was evaluated by nonparametric Spearman’s rho analysis.ResultsThe results revealed a reduction of miR-146a expression in 50% of cancerous tissue when compared with adjacent normal regions (P-value=0.127). COX-2 expression in 80% of ESCC patients was higher than in the controls (P-value=0.001). Overall, in 60% of cases, direct association was seen between microRNA-146a and COX-2 expression level (correlation coefficient= 0.438, P-value=0.011). COX2 can be considered as a diagnostic biomarker (AUC=0.834, sensitivity=72%, specificity =83%, P-value<0.0001) but miR146a cannot be considered as a diagnostic biomarker (AUC=0.553, sensitivity=88%, specificity =28%, P-value=0.453). Survival analysis by Kaplan–Meier method showed miR146a and COX2 expression can be probably considered as prognostic biomarkers for ESCC because patients with high expression of miR146a had 7 months shorter life span and patients with low expression of COX2 had 8 months shorter life span.ConclusionCOX2 expression is a diagnostic biomarker. MiR-146a and COX2 expression can probably be considered as prognostic biomarkers for survival in ESCC.

Tumor Necrosis Factor-Induced miR-146a Upregulation Promotes Human Lung Adenocarcinoma Metastasis by Targeting Merlin

Inflammation plays a key role in carcinogenesis and metastasis. This process involves the inactivation of tumor suppressor molecules, yet the molecular mechanisms by which inflammation impairs tumor suppressors are not completely understood. In this study, we show that proinflammatory signals such as tumor necrosis factor (TNF) support lung cancer metastasis by reducing the levels of the tumor suppressor Merlin through regulation of miR-146a. Immunodeficient mice inoculated with A549 cells expressing high miR-146a levels and low Merlin protein levels exhibited reduced survival, which correlated with the number of metastatic nodes formed. Accordingly, restoring Merlin protein levels inhibited metastasis and increased survival of the mice. Consistent with these results, we found that elevated miR-146a expression levels correlated with low Merlin protein levels in human lung adenocarcinoma. Furthermore, human invasive and metastatic tumors showed higher TNF and miR-146a levels, but lower Merlin protein levels than noninvasive tumors. These findings indicate that upregulation of miR-146a by TNF in lung adenocarcinoma promotes Merlin protein inhibition and metastasis. Thus, we suggest that the ratio between miR-146a and Merlin protein levels could be a relevant molecular biomarker that can predict lung cancer progression and that the TNF/miR-146a/Merlin pathway is a promising new therapeutic target to inhibit lung adenocarcinoma progression.

Predictive Value of miRNA-181a in Pediatric Acute Lymphoblastic Leukemia

Background: Acute lymphoblastic leukemia (ALL), a common pediatric malignant neoplasia, showed high relapse rate after induction therapy. Some miRNAs have been shown to regulate normal hematopoiesis and their disruption could contribute to leukemogenesis: Recently, specific miRNA, including miR-181a, was shown to be involved in the pathogenesis of ALL, serving as a biomarker for diagnosis and relapse of ALL. Aim of the Study: To evaluate miR-181a expression level as a predictive marker for children with acute lymphoblastic leukemia. Patients and Methods: 40 pediatric ALL patients were included in this study. miR-181a expression was assessed at diagnosis before start of treatment. Samples were either peripheral blood or bone marrow aspirate sample. Patients were evaluated clinically and laboratory after the induction therapy. Results: The remission rate was significantly higher in patients with high miR-181a expression compared to those with low expression (p Conclusion: The expression level of miR-181a was significantly higher in remission group than in non-remission group was and predict good response to induction therapy.

The renoprotective effect of curcumin against cisplatin-induced acute kidney injury in mice: involvement of miR-181a/PTEN axis

Background Nephrotoxicity, especially acute kidney injury (AKI), is the main dose-limiting toxicity of cisplatin. Although recent studies showed that curcumin prevented cisplatin-induced AKI effectively, further studies to understand the mechanism are required. Methods We established an AKI mouse model. Male C57BL/6 mice were assigned to three groups: saline group (control), cisplatin group (CP), and curcumin + cisplatin group (CP + Cur). The CP group received a single intraperitoneal (i.p.) injection of cisplatin, while the control group received saline. The CP + Cur group received i.p. curcumin three days before cisplatin injection and curcumin administered for another three days until the day before euthanization. Renal injury was assessed by serological and histological analysis. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) were used to detect the phosphatase and tensin homolog (PTEN), and microRNA (miR)-181a expression in the renal tissues. Bioinformatics prediction and western blotting methods validated the targets of miR-181a in vitro. Results Curcumin treatment alleviated cisplatin-induced nephrotoxicity as validated by the blood urea nitrogen (BUN) values, and histological analysis of kidneys. At the molecular level, curcumin treatment decreased miR-181a expression level, which was induced by cisplatin and restored the in vivo expression of PTEN, which was suppressed by cisplatin. We verified the direct regulation of PTEN by miR-181a in cultured human embryonic kidney 293T cells. Conclusions We showed the involvement of miR-181a/PTEN axis in the renoprotective effect of curcumin against cisplatin-induced AKI, and provide new evidence on the ability of curcumin to alleviate cisplatin-induced nephrotoxicity.