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A large number of clinical trials have proved that radiotherapy is an effective strategy for treating breast cancer (BC), but radioresistance is an urgent problem to be solved. Herein, radio-tolerant BC cell lines named MDA-MB-231/R and MCF-7/R were successfully constructed, respectively, and it was shown that microRNA-125b-5p (miR-125b-5p) was lowly expressed in radio-tolerant cells, which was associated with poor prognosis of BC patients. Furthermore, miR-125b-5p over expression could hinder cell proliferation and strengthen radio-sensitivity. Mechanistically, Breast Cancer 1 (BRCA1) was identified as a downstream protein target of miR-125b-5p in BC cells, and Kruppel-like factor 5 (KLF5) was identified as an upstream transcription factor involved in promoting miR-125b-5p expression. More importantly, over expression of KLF5 suppressed BC cell proliferation and increased its radiation sensitivity, which could be retained by miR-125b-5p downregulation. In conclusion, these findings suggested that the KLF5/miR-125b-5p/BRCA1 axis may provide new information on radiation therapy for breast cancer.

Background MicroRNAs (miRs) are small non-coding RNAs. MiR-125b has been described as being downregulated in cataract tissue when compared to a transparent lens. Objectives The aims of the study were: 1) to establish the expression of miR-125b in cataracts complicated by pseudoexfoliation syndrome (PEX), glaucoma or PEX glaucoma; and 2) to determine whether any environmental factors influence miR-125b expression. Material and methods Anterior lens capsules were obtained from 150 patients. The patients were subdivided into 1 of 4 groups: those with PEX (PEXg), those with primary open-angle glaucoma (Gg) and those with PEX glaucoma (PEXGg), plus gender-matched controls with cataracts alone (control group - Cg). Quantitative polymerase chain reaction (qPCR) expression of microRNA-125b was examined in every group. Results The mean age of the 150 patients was 75.18 years (standard deviation (SD) ±9.12 years). Our investigation indicated, for the first time, that miR-125b expression was increased 3.33 times in the PEXg (p = 0.015). The quantitative analysis of miR-125b expression conducted between combined groups of all the patients that have PEX syndrome (with or without glaucoma) and the Cg revealed a statistically significant difference (p = 0.04). Lower miR-125b expression was found in the patients who smoked compared to those who did not (p = 0.01). Conclusions Our data revealed the possible role of miR-125b in PEX syndrome development. There are 2 possible interpretations of these results: either the co-existence of PEX acts as a moderator of miR-125b expression in the anterior lens capsule, or increased expression of miR-125b can play a role in the pathogenesis of PEX.

Aim: The aim was to systematically investigate the miRNA biomarkers for early diagnosis of hepatocellular carcinoma (HCC). Materials & methods: A systematic review and meta-analysis of miRNA expression in HCC were performed. Results: A total of 4903 cases from 30 original studies were comprehensively analyzed. The sensitivity and specificity of miR-224 in discriminating early-stage HCC patients from benign lesion patients were 0.868 and 0.792, which were superior to α-fetoprotein. Combined miR-224 with α-fetoprotein, the sensitivity and specificity were increased to 0.882 and 0.808. Prognostic survival analysis showed low expression of miR-125b and high expression of miR-224 were associated with poor prognosis. Conclusion: miR-224 had a prominent diagnostic efficiency in early-stage HCC, with miR-224 and miR-125b being valuable in the prognostic diagnosis.

Background: Several studies investigated the miRNAs in cancer trying to assess the prognosis or to predict the response to certain treatment .One of these miRNAs are miR 125b , it was suggested by previous results as a good marker for prediction of aromatase inhibitors (AI) response. So, this study was conducted to assess the value using miRNA125b as a predictive factor to AI response. Patients and Methods: A total of 90 patients of postmenopausal HR+ve metastatic breast cancer who attended to medical oncology department outpatient’s clinic in SECI, Assiut university Egypt, from May 2017 to September 2018. Patients presented with metastasis at diagnosis as well as patients who relapsed only after 3 years of adjuvant hormonal treatment with SERM were included. All patients received AI as first line treatment for metastatic, miRNA 125b was isolated from peripheral blood samples and measured by using (q PCR).The response of patients was assessed by RECIST criteria and correlated with its expression levels. Results: Expression of the miR125-b was significantly higher in patients than control p= 0.000. However, no significant difference in its expression between those with single of multiple metastases or even between the site of metastases. We didn’t find also any significant difference in response p=0.648 and survival either PFS p=0.406 or OS p=0.384 between those patients with high expression vs. low expression. Conclusion: Our results suggest that miR125b could be used as a diagnostic marker as it is significantly increased in patients than control. However, we don’t recommend using miR125b as a marker to predict the AI resistance as further studies with large sample size are needed to confirm these results.

The underlying mechanisms of recurrence and metastasis of epithelial ovarian cancer (EOC) are largely unknown. In the present study, we investigated the clinical significance of microRNA-125b (miR-125b) and its role in ovarian tumorigenesis and progression. Seventy patients of EOC and paired tissues were enrolled from 2015 to 2017. qRT-PCR was used to evaluate miR-125b expression in tumor tissues and EOC cell line. Gain-and-loss function of miR-125b was achieved to explore the changes in cell biological function. We found that miR-125b expression in EOC tissues, especially in the high-grade tissues (P, Graphical abstract

Background: Mitochondria are dynamic organelles whose structure are maintained by continuous fusion and fission. During acute kidney injury (AKI) progression, mitochondrial fission in renal tubular cells was elevated, characterized by mitochondrial fragmentation. It is tightly associated with mitochondrial dysfunction, which has been proven as a critical mechanism responsible for AKI. However, the initiating factor for the disruption of mitochondrial dynamics in AKI was not well understood. Objectives: To explore the molecular mechanisms of mitochondrial disorders and kidney damage. Methods: We established cisplatin-induced AKI model in C57BL/6 mice and proximal tubular cells, and detected the expression of miR-125b by qPCR. Then we delivered miR-125b antagomir after cisplatin treatment in mice via hydrodynamic-based gene transfer technique. Subsequently, we performed luciferase reporter and immunoblotting ­assays to prove miR-125b could directly modulate mitofusin1 (MFN1) expression. We also tested the role of miR-125b in mitochondrial and renal injury through immunofluorescent staining, qPCR, and immunoblotting assays. Results: miR-125b levels were induced in cisplatin-challenged mice and cultured tubular cells. Anti-miR-125b could effectively alleviate cisplatin-induced mitochondrial fragmentation and kidney injury both in vitro and in vivo. Furthermore, miR-125b could directly regulate MFN1, which is a key regulator of mitochondrial fusion. Our study indicated that miR-125b is upregulated during cisplatin-induced AKI. Inhibition of miR-125b may suppress mitochondrial and renal damage through upregulating MFN1. This study suggests that miR-125b could be a potential therapeutic target in AKI.

With the development of miRNAs identification technology, more and more miRNAs have been discovered, and the role of miRNAs in the development of animal hair follicles has become a focus of research on hair-producing animals. In the previous experiment, compare the microRNA (miRNA) trancriptomes of goats and sheep skin using Solexa sequencing and differentially expressed miR-125b was screened. However, the mechanism of miR-125b regulating hair follicle development is not clear. Therefore, in the present study, the expression of miR-125b, MXD4 and FGFR2 in skin tissue of Fine-wool Sheep and Cashmere goats and HEK-293T cells was examined by qPCR and Western blot. Furthermore, the correlation between miR-125b and the predicted target gene (MXD4, FGFR2) was verified using the Dual-luciferase Reporter assay. We demonstrated that the expression of MXD4 and FGFR2 in Cashmere goats was significantly higher than that of Fine-wool Sheep, and the expression was opposite to that of miR-125b. miR-125b can down-regulate the levels of MXD4 and FGFR2. Dual-luciferase reporter gene assay showed that miR-125b could bind to the 3'-UTR region of target genes FGFR2 and MXD4, suggesting that MXD4 and FGFR2 were target genes of miR-125b. This study has shown that the growth and development of hair follicles in skin tissue of Fine-wool Sheep and Cashmere goats from the new regulatory levels of miRNAs, and clarified the mechanism of miR-125b and its target genes in the development of hair follicles in the skin.

Study question To investigate the expression profiles of microRNA–125b and micrpRNA–142–3p in women with endometriosis, compared with controls before surgery and at 1 month after surgery. Summary answer The over-expression of miRNA–125b and miRNA–142–3p may be involved in the aetiopathogenesis of endometriosis which is related to systemic chronic inflammation. What is known already Currently, there is no reliable non-invasive diagnostic biomarker for endometriosis. MicroRNAs (miRs) are small, non-coding RNAs that are involved in the post-transcriptional regulation of gene expression and promising biomarker candidates for noninvasive diagnosis of various diseases. Despite the small number of studies found that miRNA–125b and miRNA–142–3p have been associated with endometriosis, further evidence is therefore required from studies that examine these two miRNAs in diagnosis and even follow-up of women with endometriosis. Owing to endometriosis is a systemic chronic inflammatory disease, investigating these miRs in blood leukocytes of patients with endometriosis may illustrates the molecular mechanism of endometriosis. Study design, size, duration This is a prospective longitudinal study performed between 2018 November and 2021 February. The sample size of 42 individuals of two groups were calculated considering the power analysis (α = 0.05) with Mann-Whitney U test (effect size,d=0.8) and were calculated as 80%. Women with endometriosis (n = 21) and surgically confirmed endometriosis-free women (n = 21) were included in the study. Laparoscopy and/or laparotomy was performed to determine the presence or absence of endometriosis. Participants/materials, setting, methods Women aged 18–50 years were recruited from two tertiary hospital settings. Severity of endometriosis was assessed by the rASRM classification. Using real-time quantitative PCR, miRNA–125b and miRNA–142–3p in leukocyte were analyzed in women with endometriosis before surgery and at 1 month after surgery, compared to controls without endometriosis. The results were calculated as relative quantification values. The presumed targets of these two miRNAs were identified via 3 different target prediction algorithms:TargetScan, miRDB and DIANA-TarBase. Main results and the role of chance There were no demographic discrepancies between groups. The relative expression of miRNA–125b and miRNA–142–3p were significantly higher in women with endometriosis than in control subjects before surgery (p = 0.0001;p=0001) and at 1 month after surgery, respectively (p = 0.0001;p=0001). Despite the relative expression of miRNA–125b and miRNA–142–3p were decreased 1.75- and 2.4-fold, respectively at 1 month after surgery, we observed no significant differences in the relative expression of miRNA–125b and miRNA–142–3p between before surgery and at 1 month after surgery, respectively (p = 0.110; p = 0.910). Bioinformatic analyses of three databases showed that miRNA–142–3p expression levels were found to be closely associated with fifty-seven genes. Among these target genes, CFL2, RGL2, WASL, CRK, BNC2, CLOCK, TGFBR1, CIITA and ZNF217 were found to be associated with endometriosis. Whereas, no target gene were observed to be associated with miRNA–125b expression in common with these three databases. The ROC curves showed that the expression of miRNA–125b and microRNA–142–3p had an area under the ROC curve (AUC) of 0.77 (sensitivity 61.9%, specificity 88.1% (0.0001;95%CI 0.65–0.90)) and AUC of 0.71 (sensitivity 52.4%, specificity 73.8% (p Limitations, reasons for caution Further studies are needed to examine the expression of these miRs with a long-term follow up in order to increase their usefulness as a predictor in the clinical practice. It is required to identify the expressions levels of predicted target genes which are associated with endometriosis and regulated by miRNA–142–3p. Wider implications of the findings: Findings suggest that the over-expression of miRNA–125b and miRNA–142–3p may be potential mechanisms involved in the etiopathogenesis of endometriosis which is a systemic chronic inflammatory disease. We observed that miRNA–125b may be a more reliable biomarker than miRNA–142–3 for noninvasive diagnosis of endometriosis and even follow-up of women with endometriosis. Trial registration number 118S298

Osteoarthritis is a bone-joint disease prevalent in older people characterized by joint inflammation. In traditional Chinese medicine, polydatin plays an important anti-inflammatory role. This study analyzed the potential effects and possible internal mechanisms of polydatin on osteoarthritis. First, lipopolysaccharide-induced osteoarthritis injury was established in chondrogenic ATDC5 cells. Lipopolysaccharides significantly stimulated inflammatory injuries in ATDC5 cells as exemplified by a decrease in cell viability and an increase in inflammatory cytokine secretions including interleukin-6, tumor necrosis factor-a, and interleukin-1. Moreover, lipopolysacchrides also increased Cleaved caspase-3 and Cleaved Poly (ADP-ribose) polymerase to promote cell apoptosis. Second, polydatin showed significant protective effects against lipopolysaccharide-induced inflammatory injury, again exemplified by increased cell viability, decreased inflammatory cytokines, Cleaved caspase-3, and Cleaved poly (ADP-ribose) polymerase. Lastly, miR-125b and its binding target Rho-Associated Coiled-Coil Containing Protein Kinase 1 were closely associated with regulatory effects of polydatin against lipopolysaccharide-stimulated ATDC5 cell inflammatory injuries. Polydatin alleviated lipopolysaccharide-stimulated inflammatory injuries via the down-regulation of miR-125b. The present study concludes that polydatin plays an anti-inflammatory role in lipopolysaccharide-stimulated ATDC5 cell inflammatory injuries via the down-regulation of miR-125b.

Propranolol could repress infantile hemangioma cell growth and induce apoptosis. miR-125b could inhibit cell proliferation in some tumors. However, whether propranolol exerts its proliferation inhibition and apoptosis-promoting effect by regulating the expression of miR-125b needs to be further investigated. In tumor tissue and endothelial cells isolated from infantile hemangioma patients, we found that the expression levels of miR-125b were significantly decreased. In-vitro analysis revealed that propranolol increased the expression of miR-125b in hemangioma cells in a dose-dependent and time-dependent manner. Interestingly, it was observed that regression of miR-125b expression by its inhibitor could abrogate the effect of propranolol on hemangioma cell growth and apoptosis. In addition, our data further identified TFAP4 as a direct target of miR-125b. Collectively, our data provided evidence that propranolol may repress infantile hemangioma cell growth and promote apoptosis through upregulating the miR-125b expression, which exerted its suppression of tumor development by targeting TFAP4.

Allowing for the correlation between intermittent hypoxia and impaired cognition, we intended to figure out whether and how dementia-relevant biomolecules (e.g. miR-125b and p38MAPK) might participate in weakening of intelligence in the context of intermittent hypoxia. Within this investigation, four groups of rat models were established, including rats treated with none, 10% continuous hypoxic (CH), 10% chronic intermittent hypoxia (CIH) and 5% chronic intermittent hypoxia (CIH). Moreover, the learning, memorizing and cognitive abilities of rats were evaluated through performing morris water maze, spatial navigation test and spatial probe test, respectively. Furthermore, expressions of miR-125b and p38MAPK were determined based on reverse transcription-polymerase chain reaction (RT-PCR) or western blotting. And the luciferase reporter gene assay was prepared to validate the targeted relationship between miR-125b and p38MAPK. Consequently, our study results showed that intermittent hypoxia generated more serious cognitive impairment to rats than persistent hypoxia (P 0.05), and the rats treated with intermittent hypoxia were observed with down-regulated miR-125b expression and up-regulated phospho-p38MAPK expression (P 0.05). Furthermore, the in-vitro tests exhibited that down-regulated expression of miR-125b and phosphorylation of p38MAPK could promote apoptosis of SH-SY5Y cells, and also depress their viability and proliferation (P 0.05). The luciferase reporter gene assay also validated a targeted relationship between miR-125b and p38MAPK in SH-SY5Y cells. In conclusion, miR-125b targeting p38MAPK might be involved with modifying learning, memorizing and cognitive abilities of rats that were managed with intermittent hypoxia, which might provide theoretical basis for treating intermittent hypoxia-related dementia.

Introduction TP53 and MGMT alterations play a crucial role in glioblastoma (GB) pathogenesis. TP53 and MGMT function is affected by several pathologic mechanisms, such as point mutations or promoter methylation, which are well characterized. Expression of both genes can be regulated by other mechanisms as well, e.g., microRNAs (miRNAs). Moreover, cross-talk among various pathologic processes may occur, further affecting MGMT and TP53 functionality. Material and methods In 49 GB patients, we analyzed the possible associations between TP53 and its miRNA regulators miR-125b, miR-21, and miR-34a, as well as MGMT and its miRNA regulators miR-181d and miR-648. We evaluated the possible influence of mutational and methylation status on the pre-identified associations. Results In patients with immunohistochemistry-detected TP53 overexpression, expression levels of miR-34a and TP53 were negatively correlated (r = -0.56, p = 0.0195), and in patients with TP53 mutations, expression levels of TP53 and miR-21 were negatively correlated (r = -0.67, p = 0.0330). In patients with MGMT methylation, expression levels of MGMT were negatively correlated with miR-648 and miR-125b expression levels (r = -0.61, p = 0.0269 and r = -0.34, p = 0.0727, respectively). Conclusions Our findings demonstrate that selected miRNAs are significantly correlated with MGMT and TP53 levels, but the extent of this correlation differs regarding the TP53 and MGMT mutational and promoter methylation status.

MicroRNAs, small noncoding RNAs, not only regulate gene expression at the post-transcriptional level in a variety of physiological processes but also accompany the initiation and progression of a vast number of diseases, including dementia. While miR-125b has been shown to be aberrantly expressed in some dementia patients, its role in the pathological process remains ambiguous. Presenilin-1/2 conditional double knockout mice exhibit a range of symptoms, including impaired cognition and memory, increased tau phosphorylation, neuroinflammation, and apoptosis, and are therefore regarded as a useful dementia model. In the prefrontal cortices of double knockout mice, miR-125b was found to be abnormally increased in an age-dependent manner. We further verified the neural cell adhesion molecule (NCAM) as an miR-125b target using the dual luciferase reporter assay. The NCAM protein level was decreased when miR-125b was overexpressed (OE) in neuronal growth factor-induced differentiated PC12 cells, which further inhibited the neuronal growth factor-induced phosphorylation of glycogen synthase kinase 3 beta (GSK3β) at the Ser9 site and ultimately increased the GSK3β activity and tau phosphorylation. Moreover, on serum deprivation, high GSK3β activity in differentiated miR-125b-OE PC12 cells induced increased caspase-3 activation. Finally, adeno-associated virus-mediated miR-125b overexpression in the prefrontal cortexes of wild-type C57B/L6 mice resulted in decreased dendritic spine density. In addition, similar to the in vitro data, elevated GSK3β activity and hyperphosphorylation of the tau protein were confirmed. Taken together, our findings reveal a direct regulation of miR-125b on NCAM, which leads to further effects on downstream GSK3β activity and tau phosphorylation and may contribute to the generation of neurofibrillary tangles in neuropathological progression.

BackgroundIdentification of molecular markers, such as miRNAs is promising for the diagnosis of asthma and its clinical phenotypes. The aim of this study was to examine the changes in the expression of selected microRNAs in plasma exosomal fractions of severe asthma patients. The expression of miRNAs was determined in relation to the changes in inflammatory markers.MethodSevere asthma patients (n = 30) and healthy subjects (n = 30) were selected among the individuals referred to asthma and allergy clinic. Blood was collected from each participant to determine the serum high-sensitive C-reactive protein (hs-CRP) and total IgE. The exosomal fraction of plasma was isolated and processed for quantitation of miR-124, miR-125b, miR-133b, miR-130a and miR-125b-1-3p expression using quantitative real time-PCR (qRT-PCR).ResultsSerum hs-CRP and total IgE were significantly higher in asthma patients compared to controls. Expression of miR-124, miR-133b, and miR-130a was down-regulated in asthma patients as compared to controls (p ConclusionOverexpression of miR-125b in severe asthma which was associated with serum IgE and hs-CRP may suggest that this molecule is linked to inflammatory reactions. Up-regulation of miR-125b together with decreased expression of miR-124, miR-133b, and miR-130a may suggest that this miRNA profile is useful for diagnosis and discrimination of clinical phenotypes of asthma.

IntroductionProcalcitonin (PCT) has been reported to function as a predictive biomarker of post-operative infection. In this study, we aimed to explore the mechanisms underlying the regulation of PCT expression.Material and methods72 children diagnosed with post-operative infection and 58 children without post-operative infection were recruited in this study. Computational analysis, luciferase assay, real-time PCR, Western-blot analysis, and assays of post-operative C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) and PCT were performed to investigate the mechanisms underlying the regulation of PCT expression.ResultsMiR-125b was found to repress STAT3 expression with putative binding sites in 3’UTR of STAT3. The levels of PCT and miR-125b in non-infection group remained stable from day 0 to day 5, while the level of PCT was increased in the infection group along with a decreased level of miR-125b from day 1 to day 5. The post-operative levels of CRP and ESR in both non-infection and infection groups were evidently increased in a time-dependent manner, but the levels of miR-106b and miR-20a in both non-infection and infection groups remained stable. The area under the curve (AUC) values of PCT, CRP, ESR, miR-125b, miR-106b and miR-20a demonstrated that only miR-125b and PCT were involved in infection. Transfection with miR-125b reduced STAT3 expression, while the activation of STAT3 by lipopolysaccharide (LPS) treatment up-regulated PCT production. Finally, miR-125b down-regulated the expression of PCT by targeting STAT3.ConclusionsTaken together, we suggested that miR-125b was involved in the prognosis and diagnosis of poster-operative infection by modulating the signaling pathway of miR-125b/STAT3/PCT.

IntroductionIn this study, we validated the changes in microRNA (miRNA) expression in hypoxic human kidney 2 (HK2) cell-derived extracellular vesicles (EVs). Additionally, we investigated the mechanism by which miRNA that are derived from EVs alleviated renal ischemia-reperfusion (I/R) injury.Material and methodsHK2 cells were treated in a hypoxic chamber (1% O2) for 12 h.EVs were obtained as supernatant from ultracentrifugation and characterized. After examining 16 differentially expressed EV-miRNAs between normoxic and hypoxic conditions by RT-PCR and bioinformatics analysis, miR-125b-5p was selected for further analysis. Normoxic and hypoxic HK2 cells-derived EVs as well as EVs that were isolated from miR-125b-5p negative control (miR-NC)-transfected or miR-125b-5p mimic (miR-MI)-transfected HK2 cells were injected in mice with renal I/R injury. The degree of renal injury was assessed by periodic acid-Schiff staining, renal tubule injury score, and plasma creatinine levels. Bioinformatics analysis was performed to determine the potential target genes of differentially expressed miRNAs. RT-PCR,western blotting, luciferase reporter assay, and immunohistochemistry were performed to investigate the relationship between miR-125b-5p and the NLR family CARD domain containing 5 (NLRC5).ResultsRT-PCR revealed that from 16 differentially expressed miRNAs, four EV-miRNAs were upregulated. Animal study showed that miR-125b-5p overexpression in EVs alleviated renal I/R injury. Bioinformatics analysis predicted that NLRC5 was targeted by miR-125b-5p. Moreover, the relationship between miR-125b-5p and NLRC5 was also validated.ConclusionsThere were several miRNAs that were upregulated (including miR-125b-5p) in hypoxic HK2 cells. Hypoxia induced EVs that alleviate renal IRI, can be attributed to miR-125b-5p for targeting NLRC5.

Hepatocellular carcinoma (HCC) is a leading cause of mortality in cancer patients, but the association between miR-125b-2-3p and the onset and prognosis of HCC has not been reported in previous studies; thus, the clinicopathological implications of miR-125b-2-3p in HCC require elaboration. To examine the expression of miR-125b-2-3p in HCC, both in-house RT-qPCR and public datasets were used to calculate the standard mean difference (SMD) and the summary receiver operating characteristic (sROC). MiR-125b-2-3p was markedly lower in HCC than in non-tumor tissue as assessed by the in-house RT-qPCR which was confirmed by the integrative analysis showing the SMD being −0.69 and the area under the curve (AUC) being 0.84 based on 1,233 cases of HCC and 630 cases of non-HCC controls. To gain a overview of the clinical value of miR-125b-2-3p in HCC, all possible datasets were integrated, and lower miR-125b-2-3p levels could lead to poorer differentiation and a more advanced clinical stage of HCC. The hazard ratio (HR) of miR-125b-2-3p was also calculated using a Cox proportional hazards model, and the miR-125b-2-3p level could act as an protective indication for the survival with the HR being 0.74 based on 586 cases of HCC. Furthermore, the effect of nitidine chloride (NC), a natural bioactive phytochemical alkaloid, on the regulation of miR-125b-2-3p and its potential targets was also investigated. The miR-125b-2-3p level was increased after NC treatment, while the expression of its potential target PRKCA was reduced. Above all, a low-expressed level of miR-125b-2-3p plays a tumor suppressive role in HCC., GRAPHICAL ABSTRACT

Objective: To investigate the mechanism of long noncoding RNA (lncRNA) DANCR on the progression of hepatocellular carcinoma (HCC) cells. Methods: The expression levels of DANCR and miR-125b-5p were measured in normal hepatocytes (LO2) and HCC cell lines by quantitative reverse transcription polymerase chain reaction. HepG2 and Huh-7 cells were transfected with sh-DANCR, the negative control (sh-NC), miR-125b-5p mimic, or mimic NC or cotransfected with sh-DANCR and miR-125b-5p inhibitor. HCC cell proliferation was assessed through CCK8 and plate colony formation assay. Western blot quantified the expression levels of Bcl-2, Bax, caspase-3, and cleaved-caspase-3. Apoptotic rate was detected as well as migratory and invasive capacities. The implication of the MAPK signal pathway was assessed by detecting the expression levels of p38, ERK1/2, JNK, p-p38, p-ERK1/2, and p-JNK. Interactions between DANCR and miR-125b-5p were detected by dual luciferase reporter assay. Results: In HCC cells, DANCR was highly expressed and miR-125b-5p was decreased. sh-DANCR or miR-125b-5p mimic stimulation reduced HepG2 or Huh-7 cell progression while promoted cell apoptosis evidenced by increased apoptotic rate, elevated levels of Bax and cleaved-caspase-3, and decreased Bcl-2. Moreover, the migration rate and invasiveness of HCC cells were also inhibited by sh-DANCR and miR-125b-5p mimic. Levels of p-p38/p38, p-ERK1/2/ERK1/2, and p-JNK/JNK were suppressed by sh-DANCR and miR-125b-5p mimic. LncRNA DANCR negatively targeted and directly bound to miR-125b-5p. Knockdown of miR-125b-5p could reverse the inhibitory effects of sh-DANCR on HCC cells. Conclusion: In HCC cells, lncRNA DANCR sponges miR-125b-5p and activates MAPK pathway, thus facilitating HCC cell progression.

MicroRNAs (miRNAs) are small and highly conserved non‐coding RNAs that silence target mRNAs, and compelling evidence suggests that they play an essential role in the pathogenesis of human diseases, especially cancer. miR‐125b, which is the mammalian orthologue of the first discovered miRNA lin‐4 in Caenorhabditis elegans, is one of the most important miRNAs that regulate various physiological and pathological processes. The role of miR‐125b in many types of cancer has been well established, and so here we review the current knowledge of how miR‐125b is deregulated in different types of cancer; its oncogenic and/or tumour‐suppressive roles in tumourigenesis and cancer progression; and its regulation with regard to treatment response, all of which are underlined in multiple studies. The emerging information that elucidates the essential functions of miR‐125b might help support its potentiality as a diagnostic and prognostic biomarker as well as an effective therapeutic tool against cancer., miR‐125b is an important microRNA in cancer. It is deregulated in different types of cancer where it acts as an oncogene and/or a tumour suppressor in a context‐dependent manner. miR‐125b interacts with multiple upstream regulators and downstream targets, which are involved in a variety of signalling pathways and cellular processes during tumourigenesis and cancer progression. It also exerts as a key regulator with regards to the response of cancer cells to chemotherapy. miR‐125b holds a great potential in clinical cancer diagnosis, prognosis and therapies. Created with BioRender.com.

Background: MicroRNAs (miRNAs) have been shown to play a key role in regulating the progression of cervical cancer (CC). This study aimed to develop a circulating miRNA-based molecular signature for the diagnosis and prognosis prediction of early-stage CC. Methods: This study included 112 patients diagnosed with early-stage CC, 45 patients confirmed with cervical intraepithelial neoplasia (CIN) and 90 healthy subjects. Compared to the normal controls, the expression level of miR-21 was increased, while the levels of miR-125b and miR-370 were decreased in CC in both GSE30656 and The Cancer Genome Atlas (TCGA) cohort. The expression levels and diagnostic value of these candidate miRNAs were then validated through qRT-PCR. Their diagnostic and prognostic values for early-stage CC were further explored. Results: Compared to the patients with CIN and healthy subjects, serum miR-21 was increased, while serum miR-125b and serum miR-370 were reduced in early-stage CC. In addition, combining these molecules yielded good performance for differentiating early-stage CC from CIN or healthy subjects. Moreover, strong association was found between serum miR-21 and lymph node metastasis (LNM) as well as recurrence-free survival of early-stage CC. Similar observations were found for serum miR-125b and serum miR-370. Patients with simultaneously high serum miR-21 + low serum miR-125b + low serum miR-370 suffered a high risk of LNM and recurrence, while those with low serum miR-21 + high serum miR-125b + high serum miR-370 had little risk for LNM and recurrence. Conclusions: Combining serum miR-21, miR-125b and miR-370 as a miRNA-based signature is promising for the detection and prognosis prediction of early-stage CC.

BackgroundDifferent gastric cancer (GC) subtypes usually possess distinct clinical outcomes. The function of miR-194 in gastric cancer remains unclear and controversial. This study aimed to identify potential microRNAs that differentially expressed in subtypes and to elucidate the molecular mechanisms of miR-194 in GC.MethodsComprehensive miRNA expression analysis was performed using the available miRNA-seq data from TCGA stomach cancer cohort. The preferences of miR-194 in regulating target genes were determined by RNA sequencing studies. The function of miR-194 in GC was explored in GC cell lines by performing qRT-PCR assays, western blot assays, cell proliferation assays, luciferase report assays and flow cytometry assay.ResultsIn this study, we identified a series of miRNAs that can serve as prognostic biomarkers for GC. Among them, miR-100, miR-125b, miR-199a and miR-194 were the 4 most promising prognostic biomarkers in GC due to their significant associations with various clinical characteristics of patients. MiR-100, miR-125b and miR-199a predicted poor prognosis in GC, while miR-194 predicted favorable prognosis in GC. Besides, we provided the first comprehensive transcriptome analysis about miR-194 in GC. The results showed that miR-194 tended to regulated target genes by binding on their 3' untranslated regions in a 7-mer-A1 or 7-mer-m8 or 8-mer manner. The KEGG pathway analysis showed that cell cycle was one of the most affected pathways by miR-194 in GC. Moreover, CCND1 was proved to be a novel target gene of miR-194 in GC. Additionally, the downregulation of CCND1 by miR-194 in GC further led to cell growth inhibition and cell cycle arrest. ConclusionsMiR-100, miR-125b, miR-199a and miR-194 could serve as prognostic and diagnostic biomarkers for GC. MiR-194 might suppress GC cell growth mainly through targeting CCND1 and induction of cell cycle arrest.

Arteriosclerosis occlusive (ASO) is a manifestation of atherosclerosis (AS), and it is one of the main causes of lower limb ischemic necrosis in the elderly. Excessive proliferation and migration of vascular smooth muscle cells is one of the main causes of ASO, but the mechanism is unclear. MiR-125b is a tumor suppressor, which can inhibit the proliferation and migration in various kinds of tumors. At the same time, we find miR-125b is the most obviously down-regulated microRNA in ASO. Therefore, we explore the role of miR-125b in ASO. This study shows that up-regulated miR-125b inhibits the proliferation of vascular smooth muscle cells through CCK8 assay and Brdu assay. Meantime, the up-regulation of miR-125b restricts the migration of vascular smooth muscle cells through trans-well assay and wound healing assay. Furthermore, this study finds that up-regulated miR-125b inhibits the expression level of angio-associated migratory cell protein (AAMP) and serum response factor (SRF), which may be an important way for miR-125b to regulate the proliferation and metastasis of smooth muscle cells.

Downexpression of miRs was associated with tumor development, progression, and metastasis. This study explored the serum levels of miR-125b in patients with epithelial ovarian cancer (EOC) and to assess its diagnostic value and monitor treatment responses for patients with EOC. A total of 379 individuals were recruited and assigned to the study groups. RT-qPCR analysis was performed to confirm the association of serum miR-125b levels with tumor stages and treatment responses. The median serum levels of miR-125b in patients with EOC were significantly lower than that of other controls (P

To explore the potential effect of miR-125b on p53-mediated regulation of Bax/Cytochrome C/Caspase-3 apoptotic signaling pathway in rats with cerebral ischemia-reperfusion (CIR) injury. Sprague-Dawley (SD) rats were used to conduct CIR injury and injected with miR-125b mimic/inhibitor or p53 inhibitor (Pifithrin-α, PFT-α). Dual-luciferase reporter gene assay was used to analyze the targeting relationship between miR-125b and p53. Longa scoring and Triphenyl tetrazolinm chloride (TTC) staining were used to test the neurologic function and determine infarct size, respectively. Hematoxylin-eosin (HE) and Nissl's stainings were conducted to observe the morphology of cortical neurons. Neuronal nuclei (NeuN) expression was detected by immunohistochemical staining. QRT-PCR was performed to detect the expressions of miR-125b and p53. TUNEL staining and Western blotting was used to determine neuronal apoptosis and expressions of Bax/Cytochrome C/Caspase-3 signaling pathway-related proteins, respectively. Our results showed that miR-125b could directly target p53. As observed, overexpression of miR-125b could obviously reduce the neurological score, infarct size, and brain water content after CIR in rats, which also improved the morphology of cortical neurons, increased the number of neurons, reduced neuronal apoptosis, and inhibited the expressions of Bax/Cytochrome C/Caspase-3 pathway. Moreover,the similar results were observed in rats with CIR after injected with PFT-α. But no significant differences in each index were found in CIR group and CIR + anti-miR-125b + PFT-α group.MiR-125b exerts protective effects on CIR injury through inhibition of Bax/Cytochrome C/Caspase-3signaling pathway via targeting p53, which is likely to be a promising treatment for CIR.3'-UTR: 3-untranslated region; CIR: cerebral ischemia-reperfusion; CIS: cerebral ischemic stroke; PFT-α: Pifithrin-α; PVDF: polyvinylidene fluoride; SD: Sprague-Dawley; TBST: tris buffered saline with tween. TTC staining: Triphenyl tetrazolinm chloride staining; TUNEL: Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling.

Breast cancer is a major leading cause of cancer death in women in almost all countries, including developing countries. Despite the significant progress that has been made in cancer detection and ...

BACKGROUND MircroRNA (MiRNA) levels are associated with disease pathophysiology and are high in plasma exosomes. Plasma exosomal miRNAs serve as potential therapeutic targets and diagnosis biomarkers in some diseases but few studies have examined them in Ischemic Stroke (IS). Therefore, we explored the potential predictive value of plasma exosomal miR-422a and miR-125b-2-3p in different IS phases (acute and subacute phases). METHODS Fifty-five IS patients and 25 age and sex matched healthy controls were recruited. Patients were classified into two groups: 27 patients in acute phase (days 1-3) and 28 patients in subacute phase (days 4-14). The plasma exosomal levels of miR-422a and miR-125b-2-3p were examined via quantitative real-time polymerase chain reaction (qRT-PCR). The Areas Under the Curve (AUC) of the Receiver Operating Characteristic (ROC) curve were constructed to evaluate the diagnostic accuracy of these miRNAs in IS. RESULTS The expression levels of plasma exosomal miR-422a and miR-125b-2-3p were significantly decreased in the subacute phase group (P

// Chia-Yen Dai 1, 2, 3, 4, 5, 6, * , Yi-Shan Tsai 1, * , Wen-Wen Chou 1 , Tawei Liu 1 , Chung-Feng Huang 1, 2, 4 , Shu-Chi Wang 3 , Pei-Chien Tsai 1 , Ming-Lun Yeh 1, 4, 5 , Ming-Yen Hsieh 1 , Ching-I Huang 1, 5 , Shang-Yin Vanson Liu 7 , Jee-Fu Huang 1, 4 , Wan-Long Chuang 1, 4, 6 and Ming-Lung Yu 1, 2, 3, 4, 5, 6, 8 1 Hepatobiliary Division, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan 2 Department of Occupational and Environmental Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan 3 Health Management Center, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan 4 Faculty of Internal Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan 5 Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan 6 Center for Infectious Disease and Cancer Research, Kaohsiung Medical University, Kaohsiung, Taiwan 7 Department of Marine Biotechnology and Resources, National Sun Yat-Sen University, Kaohsiung, Taiwan 8 Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung, Taiwan * These authors contributed equally to this work Correspondence to: Ming-Lung Yu, email: fish6069@gmail.com Keywords: miR-125b; HCV; IL-6; STAT3; HCV Received: July 28, 2017 Accepted: December 01, 2017 Published: January 10, 2018 ABSTRACT Background/Aims: MicroRNA-125b (miR-125b) has been found to regulate inflammation and acts as an oncogene in many cancers. The mechanisms of miR-125b expression during hepatitis C virus (HCV) infection remain to be clarified. The present study aims to identify the factors that might regulate miR-125b expression in HCV infection. Results: High expression of miR-125b was found to correlate with HCV infection in replicon cells and in sera from HCV-infected patients, whereas the miR-125b inhibitor reduced HCV gene expression. The interleukin 6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) pathway plays an inducible effect on miR-125b gene expression. STAT3 siRNA or inhibitor could reduce HCV replication. Materials and Methods: HCV replicon cells Con1 (type 1b) and Huh7/Ava5 (type 1b) were treated with 17-hydroxy-jolkinolide B (HJB) or STAT3 siRNA. Cell viability assay and Renilla Luciferase Assay were used. Fragments of the miR-125b-1 promoter were constructed for the luciferase reporter assay. PSMB8, PSMB9, miR-125b-1, and miR-125b-2 expression was determined using TaqMan ® Gene Expression Assays. Western blot analysis was performed to assess protein abundance. Conclusions: This study elucidates a novel pathway for miR-125b in the pathogenesis of chronic HCV infection and suggests it as a possible target for treating HCV infection.

Mastitis is a complex inflammatory disease caused by pathogenic infection of mammary tissue in dairy cows. The molecular mechanism behind its occurrence, development, and regulation consists of a multi-gene network including microRNA (miRNA). Until now, there is no report on the role of miR-125b in regulating mastitis in dairy cows. This study found that miR-125b expression is significantly decreased in lipopolysaccharide (LPS)-induced MAC-T bovine mammary epithelial cells. Also, its expression is negatively correlated with the expression of NF-κB inhibitor interacting Ras-like 2 (NKIRAS2) gene. MiR-125b target genes were identified using a double luciferase reporter gene assay, which showed that miR-125b can bind to the 3' untranslated region (3' UTR) of the NKIRAS2, but not the 3'UTR of the TNF-α induced protein 3 (TNFAIP3). In addition, miR-125b overexpression and silencing were used to investigate the role of miR-125b on inflammation in LPS-induced MAC-T. The results demonstrate that a reduction in miR-125b expression in LPS-induced MAC-T cells increases NKIRAS2 expression, which then reduces NF-κB activity, leading to low expression of the inflammatory factors IL-6 and TNF-α. Ultimately, this reduces the inflammatory response in MAC-T cells. These results indicate that miR-125b is a pro-inflammatory regulator and that its silencing can alleviate bovine mastitis. These findings lay a foundation for elucidating the molecular regulation mechanism of cow mastitis.

Weifeng Liu,1–3,* Jie Hu,1,2,* Kaiqian Zhou,1,2,* Feiyu Chen,1,2 Zheng Wang,1,2 Boyi Liao,1,2 Zhi Dai,1,2 Ya Cao,4 Jia Fan,1,2,5 Jian Zhou1,2,5,6 1Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, China; 2Key Laboratory of Carcinogenesis and Cancer Invasion, Fudan University, Ministry of Education, Shanghai, China; 3Department of Hepatobiliary and Pancreatic Surgery, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China; 4Cancer Research Institute, Central South University, Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Changsha, China; 5Institute of Biomedical Sciences, Fudan University, Shanghai, China; 6Shanghai Key Laboratory of Organ Transplantation, Shanghai, China *These authors contributed equally to this work Abstract: Hepatocellular carcinoma (HCC) is the sixth most common cancer worldwide with high mortality. Circulating miRNA has been demonstrated as a novel noninvasive biomarker for many tumors. This study aimed to investigate the potential of circulating miR-125b as a prognostic marker of HCC. Exosomes were extracted from serum samples collected from two independent cohorts: cohort 1: HCC (n=30), chronic hepatitis B (CHB, n=30), liver cirrhosis (LC, n=30); cohort 2: HCC (n=128). We found that miR-125b levels were remarkably increased in exosomes compared to those in serum from patients with CHB, LC, and HCC (P

microRNA-125b has been reported to play an novel biological function in the progression and development of several kinds of leukemia. However, the detail role of miR-125b in acute myeloid leukemia (AML) is remains largely unknown. The present study aimed to investigate the biological role of miR-125b in AML and the potential molecular mechanism involved in this process. Our results showed that overexpression of miR-125b suppressed AML cells proliferation, invasion and promotes cells apoptosis in a dose-dependent manner, while the miR-NC did not show the same effect. In addition, miR-125b induced AML cells G2/M cell cycle arrest in vitro. Overexpression of miR-125b resulted in a significant decrease of the expression of p-IκB-α and inhibition of IκB-α degradation, and the nuclear translocation of NF-κB subunit p65 was abrogated by miR-125b simutaneously. To further verify that miR-125b targeted NF-κB signaling pathway, the NF-κB-regulated downstream genes that were associated with cell cycle arrest and apoptosis was also determined. The results showed that, miR-125b also affect NF-κB-regulated genes expression involved in cell cycle arrest and apoptosis. In conclusion, the present work certificates that miR-125b can significantly inhibit human AML cells invasion, proliferation and promotes cells apoptosis by targeting the NF-κB signaling pathway, and thus it can be viewed as an promising therapeutic target for AML.

// Elena Vendramini 1, 2, 3 , Marco Giordan 1 , Emanuela Giarin 1 , Barbara Michielotto 1 , Grazia Fazio 4 , Gianni Cazzaniga 4 , Andrea Biondi 4 , Daniela Silvestri 4 , Maria Grazia Valsecchi 5 , Martina U. Muckenthaler 6 , Andreas E. Kulozik 6 , Valter Gattei 7 , Shai Izraeli 2, 3 , Giuseppe Basso 1 and Geertruy te Kronnie 1 1 Department of Women’s and Children’s Health, University of Padova, Padova, Italy 2 Edmond and Lily Safra Children’s Hospital, Sheba Medical Center, Ramat Gan, Israel 3 Tel Aviv University, Tel Aviv, Israel 4 Centro Ricerca Tettamanti, Clinica Pediatrica, University of Milano-Bicocca, Monza, Italy 5 School of Medicine and Surgery, University of Milano-Bicocca, Milano, Italy 6 Department of Pediatric Oncology Hematology, University of Heidelberg, Heidelberg, Germany 7 Clinical and Experimental Onco-Hematology Unit, Centro di Riferimento Oncologico, I.R.C.C.S., Aviano (PN), Italy Correspondence to: Elena Vendramini, email: elena.vendramini@gmail.com Keywords: B cell precursor acute lymphoblastic leukemia, ERG aberrations, noncoding RNAs, miR-125, SNORD116 Received: June 29, 2016 Accepted: March 04, 2017 Published: March 21, 2017 ABSTRACT ERG-related leukemia is a B cell precursor acute lymphoblastic leukemia (BCP ALL) subtype characterized by aberrant expression of DUX4 and ERG transcription factors, and highly recurrent ERG intragenic deletions. ERG-related patients have remarkably favorable outcome despite a high incidence of inauspicious IKZF1 aberrations. We describe clinical and genomic features of the ERG-related cases in an unselected cohort of B-other BCP ALL pediatric patients enrolled in the AIEOP ALL 2000 therapeutic protocol. We report a small noncoding RNA signature specific of ERG-related group, with up-regulation of miR-125b-2 cluster on chromosome 21 and several snoRNAs in the Prader-Willi locus at 15q11.2, including the orphan SNORD116 cluster.

IntroductionMicroRNA-125b has been found to be down-regulated in many types of malignant tumours and diseases with excessive proliferation of keratinocytes, such as cutaneous squamous cell carcinoma and psoriasis. Cholesteatoma, which is mainly composed of keratinocytes, also has characteristics of abnormal proliferation similar to a malignant tumour. However, the expression and regulatory mechanisms of miR-125b and its downstream genes in cholesteatoma have not been clarified.Material and methodsReal time fluorescence quantitative PCR was applied to detect the expression of miR-125b in the cholesteatoma and corresponding retroauricular skin. Immunohistochemical staining and western blot were used to detect signal transducers and activators of transcription 3 (STAT3) and the downstream gene cyclinD1, survivin, and vascular endothelial growth factor (VEGF) in the cholesteatoma and corresponding retroauricular skin. The targeted regulatory relationship between miR-125b and STAT3 was confirmed by dual luciferase reporter assay. Proliferation and apoptosis of transfected HaCaT cells were detected by MTS, cell cycle, and apoptosis assays.ResultsWe observed down-regulation of miR-125b and up-regulation of STAT3, cyclinD1, survivin, and VEGF in cholesteatoma tissues. STAT3 was a direct target gene of miR-125b. Inhibition of miR-125b enhanced STAT3 and its downstream genes expression, promoted HaCaT cell proliferation, and inhibited apoptosis.ConclusionsThe results of this study demonstrate that miR-125b can influence the growth of cholesteatoma by targeting STAT3 and its downstream genes, including cyclinD1, survivin, and VEGF, thus providing an opportunity to establish new medical therapy strategies and facilitating further study of the pathogenesis of cholesteatoma.

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Objective This study aimed to investigate the correlation of miR‐125a/b expressions with disease risk, progression, and prognosis of sepsis. Methods MiR‐125a/b expressions and inflammatory cytokines were detected by RT‐qPCR and ELISA assays in plasma samples from 120 sepsis patients. Besides, blood biochemical indexes, disease severity scores, and in‐hospital mortality of sepsis patients were recorded. Meanwhile, miR‐125a/b expressions in plasma from 120 health controls (HCs) were also detected by RT‐qPCR. Results MiR‐125b was elevated in sepsis patients compared with HCs, and ROC curve revealed that miR‐125b could well distinguish sepsis patients from HCs with AUC 0.658. MiR‐125b positively correlated with APACHE II score, SOFA score, Scr, CRP, PCT, TNF‐α, and IL‐6 levels. Most interestingly, miR‐125b was greatly decreased in survivors compared with non‐survivors, and multivariate analysis revealed that miR‐125b independently predicted higher mortality risk in sepsis patients. Besides, miR‐125a showed no significant correlation with sepsis risk, disease severity, or prognosis. Conclusion MiR‐125b but not miR‐125a is upregulated and exhibits a trend to correlate with enhanced disease severity, inflammation, and increased mortality in sepsis patients.