Expression Levels Of miR-222 Research Articles

Role of miR-138 on Radio Sensitivity of Gastric Carcinoma Cells and Its Molecular Mechanism

To explore FOXP4 and miR-138 in gastric carcinoma (GC) and its related mechanisms. Sixty-eight GC patients from January 2018 to January 2019 were selected as group A, and 66 healthy people as group B. GC cells and gastric mucosal epithelial cells were purchased, sh-FOXP4, si-FOXP4, NC, miR-138-inhibitor, and miR-138-mimics were transfected into MKN-45 and NCI-N87 cells. The FOXP4 and miR-138 expression levels in samples were tested by qRT-PCR, and N-cadherin, vimentin, Fibronectin, Slug, E-Cadherin and Y-catenin downstream proteins in cells were detected by WB. The proliferation was tested by MTT assay, invasion was tested by Transwell assay, and apoptosis was detected by flow cytometry assay. F0XP4 was highly expressed in GC, miR-138 was poorly expressed in GC, and AUC of FOXP4 and miR-138 was > 0.8. FOXP4 and miR-138 were tied to the age, gender, tumor invasion, differentiation degree, tumor location and TNM stage of GC patients. Silent expression of FOXP4 and overexpression of miR-138 can promote apoptosis, inhibit cell growth and epithelial-mesenchymal transition (ETM). The two also can inhibit N-cadherin, vimentin, Fibronectin and Slug proteins, and promote E-Cadherin and y-catenin. Dual luciferase report confirmed that FOXP4 and miR-138 had targeted relationship. In terms of radiological parameters, the SF2 and D0 (Gy) values of transfected miR-138mimic+pcDNA3. 1-FOXP4 were dramatically higher than those of transfected miR-138mimic+pcDNA3. 1 cells, and SER was lower than those of transfected miR-138 mimic+ pcDNA3. 1 cells (P<0.05), suggesting FOXP4 partially adjusted the radiosensitization role of miR-138. In conclusion, miR-138 can regulate EMT of GC cells by adjusting F0XP4 and enhance radiosensitivity of those cells.

Abstract P456: High Glucose Suppresses Cardiomyocyte Progenitor Cell Regenerative Capacity And The Role Of Mir-195/ezh2 Crosstalk In Gestational Diabetes Mellitus

Introduction: Gestational diabetes mellitus (GDM) is associated with a five-fold increase in congenital heart defects. It is critical to determine the biological effects of diabetes mellitus (DM) in vivo and high glucose in vitro on neonatal cardiomyocyte progenitor cells (nCPCs) to maximize their regenerative potential. In the present study we seek to investigate the roles of Mir-195 and its hypothesized target gene, enhancer of zeste homolog 2 (Ezh2), in GDM. Hypothesis: We predict that high glucose is associated with decreased cellular proliferation, viability and increased senescence through oxidative stress. We also hypothesize that expression of Mir-195 will be higher in DM-nCPCs and inhibit cell proliferation via Ezh2 silencing. Methods: We subjected nCPCs in vitro to increasing glucose concentrations; cellular proliferation, migration, reactive oxygen species generation, and apoptosis were assessed using cell counting kit-8, wound healing, dihydroethidium, and annexin assays respectively. Our in vivo experiments involved injecting four-week old female mice with streptozocin. After pairing diabetic mice with non-diabetic male mice, timed embryos at E14.5 were evaluated for viability, proliferation and characterization. mRNA expression levels of Mir-195 and Ezh2 protein levels were detected using RT-qPCR and western blot analysis respectively. Lipofectamine transfection, with siRNA inhibiting Mir-195, was performed on c-kit+ cardiac stem cells obtained from diabetic mothers. Results: We found that subjecting nCPCs in vitro to increased glucose concentration led to increased % cell death, decreased proliferation and expression of paracrine factors indicating poorer secretome quality from these cells. Our in vivo models showed that maternal diabetes impedes prenatal development as decreased expression of c-kit+/Lin- cells and ISL1+ cells and increased DHE positivity were seen in DM-nCPCs at E14.5. Expression of Mir-195 was higher in DM-nCPCs but Ezh2 mRNA and protein expression levels were significantly decreased. siRNA inhibition of Mir-195 revealed higher EZH2 expression in c-kit+ cells and concomitant increase in regenerative capacity. Conclusion: In conclusion, the viability of DM-nCPCs both in vivo and in vitro is decreased compared to NDM-nCPCs suggesting decreased postnatal regenerative capacity. Mir-195 is associated with increased apoptosis and decreased proliferation of nCPCs via abrogation of the protective effects of EZH2.

Expression profiles of miR-196, miR-132, miR-146a, and miR-134 in human colorectal cancer tissues in accordance with their clinical significance : Comparison regarding KRAS mutation.

Colorectal cancer (CRC) is among the most widespread malignancies in the world. MicroRNA (miRNA) has been identified as an important modulator of the biological processes of the cells. This group of noncoding RNAs also has apivotal function in the growth and development of human cancers, including CRC. Among these miRNAs, miR-196, miR-132, miR-146a, and miR-134 have fundamental impacts on the regulation of cancers. The current study aimed to investigate the involvement of these miRNAs in CRC patients. In this study, 50pairs of tumor and tumor margin samples of CRC patients were investigated to assess the expression levels of miR-196, miR-132, miR-146a, and miR-134 in this cancer. For this purpose, firstly, quantitative real-time PCR (qRT-PCR) was applied. Also, KRAS mutation and clinicopathological characteristics of the CRC patients were analyzed in the study groups. The findings demonstrated the overexpression of miR-196 (P-value = 0.0045) and miR-146a (P-value = 0.0033) in tumor tissues compared to controls. Conversely, the expression levels of miR-132 (P-value = 0.00032) and miR-134 (P-value < 0.0001) were downregulated in tumor tissues. Also, miR-146a was the only miRNA with significant expression change in the case of the KRAS gene mutation. Interestingly, the expression ratio of these miRNAs was significantly associated with some of the clinicopathological features of the patients, such as lymph node and distant metastases. Our data demonstrated that these miRNAs appear to be promising novel biomarkers for early diagnosis of CRC and may pave the way for the future establishment of novel therapeutic options for CRC.

High expression of miR-195 is related to favorable prognosis in cytogenetically normal acute myeloid leukemia

Acute myeloid leukemia (AML) is a heterogeneous blood disease with poor treatment effect and high recurrence rate. With the deepening of non-coding RNA research, more and more miRNAs have been found to participate in various physiological processes of tumors. In this study, we tried to find the miRNA related to the prognosis of AML. Collect gene expression data and clinical information of AML patients in the Cancer Genome Atlas database for statistical analysis. The expression level of miR-195 of each patient was standardized by logCPM and then produced as a box plot according to subtype classification. TargetScan was used to predict the target genes of miR-195, and these genes were subjected to GO pathway enrichment analysis by Metascape. Differential miRNAs were screened through the DESeq2 package in the R language. Survival rates were estimated using the Kaplan-Meier method and the log-rank test. The multivariate Cox proportional hazard models of EFS and OS were established. We found that the expression of miR-195 was the lowest in cytogenetically normal (CN-) AML, and high expression of miR-195 only promoted the prognosis of chemotherapy-only CN-AML patients (EFS: P = 0.016; OS: P = 0.035). Multivariate analysis showed that miR-195high was a favorable and independent factor for CN-AML (both P < 0.05). Further analysis showed that miR-195 may affect signal transduction through ANHAK2 in AML. We found that high expression of miR-195 can increase prognosis time of chemotherapy-only CN-AML patients, providing a new possibility for treatment.

Identification of microRNAs responsive to shear loading in rat skin.

Pressure injuries (PIs) are localised skin injuries that result from pressure with or without shear force. Shear force is more destructive than pressure in clinical settings. Therefore, determining the critical external forces is important for selecting the appropriate care to prevent PIs. To quantitatively distinguish pressure and shear loading with high specificity, we focused on microRNAs (miRs). This study aimed to identify the miRs that are distinguishable between pressure with and without shear loading in rat skin. Microarray analysis identified six candidate miRs from the comparisons among the pressure, shear, and unloaded groups. We analysed the expression levels of the candidate miRs in the process of PI development using real‐time reverse transcriptase polymerase chain reaction. In the pressure and shear groups, miR‐92b expressions at 6 hours after loading were 2.3 ± 1.3 and 2.9 ± 1.0, respectively, which were significantly higher than those in the control group (P = .014 and .004, respectively). miR‐877 expression at 6 hours after loading was significantly increased only in the shear group (2.8 ± 0.9) compared with the control group (P = .016). These results indicate that miR‐92b and miR‐877 are promising biomarkers to determine for which external force healthcare professionals should intervene.

MiR-221 Expression Level Correlates with Insulin-Induced Doxorubicin Resistance in MCF-7 Breast Cancer Cells.

Objective Insulin induces anti-cancer drugs resistance in tumor cells. However, the mechanism by which insulin induces its drug resistance effects is not clear. In the present study, the expression of miR-221 in insulin-treated MCF-7 cells in response to the anti-cancer drug doxorubicin, was investigated. Materials and Methods In this experimental study, cell viability was evaluated using MTT (3-[4,5 dimethylthiazol-2- yl]-2,5-diphenyl tetrazolium bromide) assay. The expression level of miR-221 was determined by real time polymerase chain reaction (RT-PCR). Furthermore, the expression of insulin receptor (IR) and cleaved caspase-3 protein was assessed by Western blotting. Results The results showed that treatment of the MCF-7 cells with insulin reduced the anti-cancer effects of doxorubicin. Viability of naive and insulin-treated cells following doxorubicin (DOX) treatment was 62.9 ± 5.7% and 79 ± 7.2%, respectively. Furthermore, the expression of miR-221 in insulin-treated cells was significantly increased (2.6 ± 0.37-fold change) as compared with the control group. A significant decrease (26%) in the expression of caspase-3 protein and a significant increase (24%) in IR were observed in insulin-induced drug resistant MCF-7 cells as compared to the naive cells. ConclusionTogether, the data showed a positive correlation between the expression of miR-221 and IR expression, but a negative correlation with caspase3 expression, in insulin-induced drug resistant MCF-7 breast cancer cells. This could suggest a new mechanism for the role of miR-221 in cancer drugs resistance induced by insulin.

MiR-451 on Myocardial Ischemia-Reperfusion in Rats by Regulating AMPK Signaling Pathway.

Myocardial infarction is the main cause of death in patients with coronary heart disease. At present, the main method to treat cardiovascular disease is perfusion therapy. Myocardial ischemia-reperfusion will inevitably lead to reperfusion injury, which is also a major problem in the treatment of cardiovascular diseases. It has been reported that mir-451 in microRNA family participates in the protection of myocardial ischemia-reperfusion by regulating AMPK. The aim of this study was to investigate the effect of mir-451 on myocardial ischemia-reperfusion in rats by regulating AMPK signaling pathway. Sixty adult male rats were selected to establish myocardial ischemia-reperfusion animal model by ligating and loosening coronary artery. The expression level of mir-451 was regulated by injection of mir-451 virus vector and antibody, and the effect of increased or decreased mir-451 expression level on the activity of AMPK signaling pathway was detected. The myocardial infarct area and apoptosis rate of myocardial tissue were detected after 75 min ischemia-reperfusion. The results showed that when the expression level of mir-451 decreased by 15.7%, the activity index of AMPK signaling pathway was increased by 18.3%, the infarct area was reduced by 22.4%, and the apoptosis rate of myocardial cells was decreased by 25.2%. At the same time, the pathological structure of myocardial tissue was improved. Therefore, mir-451 is an inhibitor gene of AMPK signaling pathway. Reducing the expression of mir-451 can enhance the activity of AMPK signal pathway, and the increase of AMPK signal pathway activity is beneficial to reduce myocardial ischemia-reperfusion injury.

Abstract 1354: Deciphering the functional basis of synergy between taxanes and TAK715: A novel repurposed drug candidate in treatment-refractory aggressive prostate cancer

Abstract Metastatic castration resistant prostate cancer (mCRPC) is the most advanced and lethal stage of prostate cancer. A subset of advanced CRPC patients eventually develop an aggressive variant of prostate cancer (AVPCa), a rapidly progressive disease state with lack of AR expression (AR-ve mCRPC) and limited or no therapeutic options. To address this outstanding issue, we have previously reported using a novel computational algorithm (‘secDrug') and in vitro chemosensitivity and phenotypic assays to identify TAK715-a p38MAPK inhibitor as a synergistic partner for Taxanes/TX in treating AVPCa. Notably, in CRPC, MAPK pathway is involved in sustaining AR-independent cell proliferation. In this study, we investigated the genes, networks and molecular pathways underlining TX+TAK715 drug synergy in AVPCa. Gene expression profiling (mRNA sequencing) followed by Ingenuity pathway analysis (IPA) revealed upregulation of mitochondrial dysfunction and oxidative phosphorylation (OXPHOS) as the top dysregulated pathways following single-agent and combination treatment, which were confirmed by immunoblotting. In vitro ROS generation assay and Mitochondrial membrane potential measurement following combination treatment confirmed elevated intra-cellular ROS level (≈7 folds; indicating oxidative stress) and membrane depolarization (indicating mitochondrial dysfunction), respectively, in mCRPC lines. Furthermore, our IPA causal network analysis predicted significant upregulation of microRNA-132 and downregulation of miR-21 in response to combination treatment. Using in silico analysis on multiple GEO PCa datasets, we found low expression level of miR-132 was associated with poor clinical prognosis, transition from androgen dependent (AD) to independent (AI) stage and metastasis. miRNA 21 was significantly up regulated in the GEO PCa datasets. miRNA 21 is an AR-regulated miRNA that plays key role in nullifying the effect of castration, drive progression to AI stage, TX resistance and cellular invasiveness through down-regulation of tumor suppressor PTEN. Finally, using single-cell transcriptomic analysis we showed that cell subpopulations expressing the PCa stemness marker CD44, or drug resistant markers (CXCL8, CDK1), or epithelial-to-mesenchymal transition markers (Vimentin, TGFB1) were also very high expressors of TAK715 target genes (CSNK1D, MAPK14, MAP4K4) indicating TAK715 is potentially effective against treatment-refractory and stem-cell like subclones.Together, we conclude that TAK715+Taxane combination may be useful in curbing oncogenic progressions in AVPCa through simultaneous inhibition of multiple oncogenic factors/pathways. Currently, we are performing functional analysis of microRNA signatures of drug synergy as well as in vivo validation of the drug combinations using mouse xenograft models. Citation Format: Sayak Chakravarti, Suman Mazumder, Farnaz Hemmati, Farshad Amiri, Taraswi Mitra Ghosh, Ujjal Kumar Mukherjee, Panagiotis Mistriotis, Timothy Moore, Robert Arnold, Clayton Yates, Amit Kumar Mitra. Deciphering the functional basis of synergy between taxanes and TAK715: A novel repurposed drug candidate in treatment-refractory aggressive prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1354.

MiR-143 Targets ERK5 in Granulopoiesis and Predicts Outcome of Patients with Acute Myeloid Leukemia

Acute myeloid leukemia (AML) is a very heterogeneous disease of the hematopoietic system which is characterized by a fast progress with poor survival options. Despite intensive research standard treatment strategies rely since decades on the same chemotherapeutics, and the overall survival options still remain poor.In recent years, there is an emerging role of microRNAs (miRNAs) in the regulation of cancer development and resistance to cancer therapies. Mechanistically, miRNAs regulate gene expression through modulation of multiple target mRNAs by mRNA degradation or block of translation. It is shown that miRNAs can act as tumor suppressors and oncogenes dependent on the type of cancer. The miRNA-143/145 cluster is shown to be down-regulated in most cancers and is able to modulate tumorigenesis by targeting tumor associated genes. Several miR-143/145 targets are well described oncogenes, such as ERK5, DNMT3A or Bcl2. Interestingly, recent studies showed that the miR-143/145 cluster can regulate multidrug resistance proteins (MDR) in various cancer types and that enforced expression of miR-143/145 leads to increased sensitivity to various drugs. Besides its well-studied function mainly in solid tumors the role of the miR-143/145 cluster or miR-143 alone especially in the hematopoietic system remains largely unknown.In this study, we report a previously unrecognized function of miR-143 in granulopoiesis. Based on our observation that hematopoietic cells undergoing granulocytic differentiation exhibit increased miR-143 expression, we show that the absence of miR-143 in mice results in a reduced number of mature granulocytes but not monocytes in blood and bone marrow. Additionally, we show that overexpression or ablation of miR-143 resulted in accelerated granulocytic differentiation or block of differentiation in several in vitro systems, including leukemic cell lines and G-CSF treated CD34+ HSPCs, respectively.To identify the mRNA targets affected by miR-143 in hematopoietic cells we performed Argonaut2-RNA-Immunoprecipitation coupled with next generation sequencing. Here we could demonstrate a strong enrichment of members of the MAPK- and mTOR-pathways in cells overexpressing miR-143. Besides several potential targets of miR-143 we found ERK5 as particularly interesting since ERK5 was found to be a central mediator of cell survival, proliferation, motility, differentiation and apoptotic regulation of normal cells. To test whether miR-143 overexpression would also affect ERK5 protein abundance we performed western blot analysis of ERK5 and activated ERK5 in hematopoietic cells infected with miR-143 O/E constructs. Here we could show that miR-143 leads to reduced ERK5 protein levels in these cells. Since deregulation of both factors miR-143 as well as ERK5 have been implicated in the response to chemotherapeutic agents we analyzed the impact of miR-143 expression on treatment response. Here we show that AML patients receiving 5-azacytidine (Vidaza) and respond to the therapy had significant higher miR-143 levels than patients which did not respond. To test whether miR-143 expression would affect the response to chemotherapeutics, we overexpressed miR-143 in leukemic cells followed by Vidaza and AC220 treatment. Our results show a significant increase of apoptosis in cells overexpressing miR-143. Finally, we observed an association of high miR-143 expression levels with a higher probability of survival in two different cohorts of patients with acute myeloid leukemia. Taken together, we identified miR-143 as a relevant factor in granulocytic differentiation, which expression may be useful as prognostic factor and as new target for therapeutic strategies in AML. DisclosuresSchwind:Novartis: Consultancy.

The expression of miR-17 and miR-29a in placenta-derived exosomes in LPS-induced abortion mice model: An experimental study.

BackgroundThe expression pattern of microRNAs in placenta-derived exosomes plays a crucial role in the regulation of immune responses and inflammation at the fetal–maternal interface.ObjectiveConsidering the immunomodulatory properties of miR-17 and miR-29a, we determined their expression levels in placenta-derived exosomes in a lipopolysaccharide (LPS)-induced abortion mice model.Materials and Methods A total of 14 pregnant BALB/c mice, aged 6–8 wk, were randomly divided into two groups (n = 7/each) on the gestational day 11.5. While the mice in the experimental group were treated with LPS, those in the control group were treated with Phosphate buffered saline; 5 hr after the treatment, the placental cells were isolated and cultured for 48 hr. Then, the cell culture supernatants were collected and used for isolation of exosomes. The isolated exosomes were confirmed by western blot and scanning electron microscopy. The miRNAs were then extracted from exosomes, and cDNA synthesized. The expression levels of miR-17 and miR-29a were evaluated by quantitative real-time PCR analysis.ResultsOur results showed that the expression levels of miR-29a in placenta-derived exosomes obtained from the experimental group increased significantly compared to the control group. Also, the expression levels of miR-17 in the placenta-derived exosomes obtained from the experimental group were found to decrease; however, it did not show significant changes compared with the control group (p > 0.05).Conclusion Inflammatory reactions at the fetal–maternal interface can alter miRNAs expression patterns in placenta-derived exosomes, especially miRNAs with immunomodulatory effects such as miR-29a.

MicroRNA-9inhibitsproliferationand progression inretinoblastoma cells bytargeting PTEN.

Retinoblastoma (RB) is the most prevalent primary intraocular malignancy, which commonly occurs during infant and childhood. Our study aimed to investigate whether microRNA-9 (miR-9) could regulate RB cells and its mechanism. qRT-PCR analysis was used to detect the expression of miR-9. In addition, to detect the migration of RB cells, wound healing assay was conducted. Xenograft tumor models in nude mice were also established, in order to assess the effects of miR-9 on tumor growth. qRT-PCR, luciferase reporter assay and western blot analysis were used to detect the target of miR-9. Initially, the expression level of miR-9 was significantly decreased in the RB tissues and blood samples from patients with RB. qRT-PCR, luciferase reporter assay and western blot analysis were used to confirm that PTEN was the target genes of miR-9 and it was negatively regulated by miR-9. When the expression of miR-9 was up-regulated, the cell viability, proliferation, migration and tumor formation were significantly suppressed. Furthermore, the expression level of PTEN was decreased after transfection of miR-9 mimic. Taken together, these results indicated that miR-9 might suppress the cell viability, proliferation, migration and tumor formation in RB by inhibiting PTEN. The in vitro and in vivo experiments demonstrated that miR-9 acts as a tumor suppressor function in RB cells and might serve as novel therapeutic targets for the treatment of RB.

Overexpression of miR-124 in Motor Neurons Plays a Key Role in ALS Pathological Processes.

miRNA(miR)-124 is an important regulator of neurogenesis, but its upregulation in SOD1G93A motor neurons (mSOD1 MNs) was shown to associate with neurodegeneration and microglia activation. We used pre-miR-124 in wild-type (WT) MNs and anti-miR-124 in mSOD1 MNs to characterize the miR-124 pathological role. miR-124 overexpression in WT MNs produced a miRNA profile like that of mSOD1 MNs (high miR-125b; low miR-146a and miR-21), and similarly led to early apoptosis. Alterations in mSOD1 MNs were abrogated with anti-miR-124 and changes in their miRNAs mostly recapitulated by their secretome. Normalization of miR-124 levels in mSOD1 MNs prevented the dysregulation of neurite network, mitochondria dynamics, axonal transport, and synaptic signaling. Same alterations were observed in WT MNs after pre-miR-124 transfection. Secretome from mSOD1 MNs triggered spinal microglia activation, which was unno-ticed with that from anti-miR-124-modulated cells. Secretome from such modulated MNs, when added to SC organotypic cultures from mSOD1 mice in the early symptomatic stage, also coun-teracted the pathology associated to GFAP decrease, PSD-95 and CX3CL1-CX3CR1 signaling im-pairment, neuro-immune homeostatic imbalance, and enhanced miR-124 expression levels. Data suggest that miR-124 is implicated in MN degeneration and paracrine-mediated pathogenicity. We propose miR-124 as a new therapeutic target and a promising ALS biomarker in patient sub-populations.

Long non-coding RNA XIST regulates ovarian cancer progression via modulating miR-335/BCL2L2 axis

BackgroundX inactivation-specific transcript (XIST) is the long non-coding RNA (lncRNA) related to cancer, which is involved in the development and progression of various types of tumor. However, up to now, the exact role and molecular mechanism of XIST in the progression of ovarian cancer are not clear. We studied the function of XIST in ovarian cancer cells and clinical tumor specimens.MethodsRT-qPCR was performed to detect the expression levels of miR-335 and BCL2L2 in ovarian cancer cells and tissues. MTT and transwell assays were carried out to detect cell proliferation, migration, and invasion abilities. Western blot was performed to analyze the expression level of BCL2L2. The interaction between miR-335 and XIST/BCL2L2 was confirmed using a luciferase reporter assay.ResultsThe inhibition of XIST can inhibit the proliferation invasion and migration of human ovarian cancer cells. In addition, the miR-335/BCL2L2 axis was involved in the functions of XIST in ovarian cancer cells. These results suggested that XIST could regulate tumor proliferation and invasion and migration via modulating miR-335/BCL2L2.ConclusionXIST might be a carcinogenic lncRNA in ovarian cancer by regulating miR-335, and it can serve as a therapeutic target in human ovarian cancer.

Potential role of miR-425, miR-155 and miR-33 in Streptococcus pneumoniae pneumonia by using bioinformatics analysis and experimental validation.

Streptococcus pneumoniae (S. pneumoniae) pneumonia is the most common cause of community-acquired pneumonia (CAP). Previous studies have suggested the diagnostic potential of microRNAs (miRNAs) in infectious diseases. In the present study, we aimed to evaluate the potential role of miRNAs in S. pneumoniae pneumonia by using bioinformatics analysis and experimental validation. Gene Expression Omnibus (GEO) datasets including GSE97922 and GSE83615 were analyzed for identifying the differentially expressed miRNAs; the miRNA-target genes network was constructed by using miRNet and the targeted genes were subject to Gene Ontology enrichment, Kyoto Encyclopedia of Genes and Genomes and REACTOME pathway analysis; the miRNA and mRNA expression levels were determined by quantitative real-time PCR; protein concentrations were determined by enzyme-linked immunosorbent assay. Our results showed that miR-425, miR-155 and miR-33 were up-regulated in the serum from CAP patients when compared to healthy controls; whereas there was no significant difference in serum miR-222, miR-149, miR-186 and miR-132 expression levels between healthy controls and CAP patients. In vitro functional studies showed that lipopolysaccharides (LPS) induced the up-regulation of miR-425, miR-155 and miR-33 in RAW264.7 cells, and miR-425, miR-155 and miR-33 inhibition attenuated LPS-induced inflammatory responses in RAW264.7 cells. In conclusion, our results showed that miR-425, miR-155 and miR-33 were up-regulated in the serum from CAP patients by using bioinformatics analysis and experimental validation; furthermore, miR-425, miR-155 and miR-33 inhibition attenuated LPS-induced inflammatory responses in RAW264.7 cells. Nevertheless, our studies are still at the preliminary stages, and the detailed roles of miR-425, miR-155 and miR-33 in S. pneumoniae pneumonia still require further investigation.

An Updated Review of the Cross-talk Between MicroRNAs and Epigenetic Factors in Cancers.

Growing evidence indicates that miRs have critical activities in adjusting cellular processes, e.g., cell death, proliferation, and cell-cycle. This study aimed to provide a concise review of the recent findings regarding tumoral miRs and the cross-talk between miRs and epigenetic factors. Like the protein-coding genes, the expression levels of miRs are mediated by various transcriptional networks. Indeed, the expression of miRs could be epigenetically modulated by DNA methylation factors and histone modifiers. Furthermore, miRs can suppress critical factors, which mediate epigenetic modifications. Besides, miRs have been implicated in cancer development, metastasis, and chemo-resistance. The aberrant expression of miRs and dysregulated modulatory circuits between miRs and epigenetic factors participate in tumor progression. Identifying tumoral miRs can provide ample opportunity to overcome chemo- resistance and bring a forefront treatment for affected patients.

Upregulation of miR-3195, miR-3687 and miR-4417 is associated with castration-resistant prostate cancer

PurposeProstate cancer (PCa) is a leading cause of cancer-related death. Upon androgen-deprivation therapy, the disease may progress further to castration-resistant PCa (CRPC) with a poor prognosis. MicroRNAs (miRNAs) are small non-coding RNAs, which play crucial roles in gene regulation. The aim of our study is to find CRPC-associated miRNAs and to evaluate their functional role.MethodsIn this study, 23 benign prostatic hyperplasia (BPH), 76 primary PCa, and 35 CRPC specimens were included. Total RNA extracted from tissue sections was used for miRNA profiling on the Affymetrix GSC 3000 platform. Subsequently, stem-loop RT-qPCR analysis was performed to validate the expression levels of selected miRNAs. PCa cell lines were transfected with miRNA mimics or inhibitors to evaluate the effects on cell proliferation, cell migration and cell invasion.ResultsIn our profiling study, several miRNAs were found to be deregulated in CRPC compared to primary PCa tissue, of which miR-205 (− 4.5-fold; p = 0.0009), miR-92b (− 3.1 fold; p < 0.0001) were downregulated and miR-3195 (5.6-fold; p < 0.0001), miR-3687 (8.7-fold; p = 0.0006) and miR-4417 (5.0-fold; p = 0.0005) were most upregulated. While KLK3, miR-21 and miR-141 expression levels in androgen-treated VCaP and LNCaP cells were increased, the expression levels of miR-3687 and miR-4417 were reduced. None of the miRNAs were androgen-regulated in the AR-negative PC3 cell line. Overexpression of miR-3687 reduced cell migration and cell invasion, whilst miR-3195 enhanced cell migration.ConclusionWe have identified several novel deregulated miRNAs in CRPC tissue, including two microRNAs that are potentially involved in tumor invasion. Our data support the hypothesized involvement of miRNAs in PCa tumorigenesis and progression to CRPC. The applicability of these miRNAs as novel biomarkers for CRPC remains to be further investigated.

Effect of miR-126 on the Proliferation and Migration of Vascular Smooth Muscle Cells in Aortic Aneurysm Mice Under PI3K/AKT/mTOR Signaling Pathway.

This paper is to investigate the expression changes of Phosphatidylinositol-3 Kinase (PI3K), protein kinase B (AKT), and Mammalian Target of Rapamycin (mTOR) in Vascular Smooth Muscle Cells (VSMCs) of aortic aneurysm mice, and to analyze the mechanism of VSMCs proliferation and migration. Aortic VSMCs cells were cultured using BALB/c mice as the research object. VSMCs were identified using artificial intelligence-based digital microscopy equipment, and liposome-transfected VSMCs experiments were performed. Real-time PCR was used for the mRNA expression levels of miR-126 and Phosphatase and Tensin Homolog (PTEN). Western blot was used for the protein expression levels of PTEN, PI3K, AKT, and mTOR. The cultured cells were identified as mouse VSMCs using digital microscopes based on artificial intelligence. Compared with the normal group, the expression of miR-126 and PTEN mRNA in the model group were significantly increased and reduced, respectively. Compared with the model group, the expression level of miR-126 and PTEN mRNA in the inhibitor group were significantly reduced and increased, respectively. Compared with the model group, the expression of miR-126 and PTEN mRNA in the ursolic acid group was significantly reduced and increased, respectively. After liposome transfection, compared with the normal group, the expression of PTEN protein in the model group was significantly reduced, and the expression of PI3K protein was significantly increased. Compared with the model group, the expression of PTEN protein was significantly increased and the expression of PI3K protein was significantly decreased in the transfection group. Compared with the control group, the expression of PI3K, AKT and mTOR protein in the model group was significantly increased. Compared with the model group, the expression of PI3K, AKT, and mTOR protein in the ursolic acid group was significantly reduced. The expressions of PI3K, AKT and mTOR protein in PI3K inhibitor group and AKT inhibitor group were significantly reduced. In conclusion, ursolic acid can inhibit the proliferation and migration of VSMCs in aortic aneurysm mice through the miR-126/PTEN/PI3K/AKT/mTOR signaling pathway. Furthermore, PTEN gene and miR-126 negatively regulate PI3K/AKT/mTOR and PTEN/PI3K/AKT/mTOR signaling pathway, respectively .

Umbilical cord miRNAs to predict neonatal early onset sepsis.

ObjectiveTo determine if miRNA (miR) expression in umbilical cord blood and umbilical cord tissue differs between neonates with early onset sepsis (EOS) versus neonates without true infection.MethodsRetrospective case-control study design of human patients with EOS (n = 8), presumed sepsis (N = 12) and non-infected control patients (N = 21). Differential expression of >300 miRs was examined using the MIHS-3001ZE-miScript miRNA PCR Array Human miFinder 384HC. Expression levels of miRs were normalized using the global Ct mean of expressed miR and compared between groups. Data analysis was performed using GeneGlobe data analysis software. Ratios of over and under-expressed miRs were calculated and compared between groups using receiver operating characteristic (ROC) curves.ResultsBoth umbilical cord plasma and umbilical cord tissue revealed several miRs with differential expression with little overlap between the two specimen types. The most overexpressed miR in plasma of EOS patients was miR-211-5p and the most overexpressed in EOS cord tissue was miR-223-5p. ROC curves comparing the ratios of over and under-expressed miRs for EOS patients and controls resulted in an area under the curve of 0.787 for cord plasma (miR-211-5p/miR-142-3p) and 0.988 for umbilical cord tissue (miR-223-5p/miR-22-3p), indicating good discrimination.ConclusionsmiRs show differential expression in EOS versus non-infected controls and presumed sepsis. A ratio of over and under-expressed miRs can provide a potentially sensitive and specific diagnostic test for EOS.

Functions and Targets of miR-335 in Cancer.

MicroRNAs (miRNAs) are small non-coding RNAs (18~25 nt in length) that act as master regulators of eukaryotic gene expression. They might play an oncogenic or tumor-suppressive role in multiple cancers. In recent decades, several studies have focused on the functions and mechanisms of miR-335 in cancer. The expression level of miR-335 in tissues and cells varies with cancer types, and miR-335 has been proposed as a potential biomarker for the prognosis of cancer. Besides, miR-335 may serve as an oncogene or tumor suppressor via regulating different targets or pathways in tumor initiation, development, and metastasis. Furthermore, miR-335 also influences tumor microenvironment and drug sensitivity. MiR-335 is regulated by various factors such as lncRNAs and microRNAs. In this review, we reveal the functions and targets of miR-335 in various cancers and its potential application as a possible biomarker in prognostic judgment and treatment of malignant tumors.

Impact of RARα and miR-138 on retinoblastoma etoposide resistance

Retinoblastoma (RB) is the most common childhood eye cancer. Chemotherapeutic drugs such as etoposide used in RB treatment often cause massive side effects and acquired drug resistances. Dysregulated genes and miRNAs have a large impact on cancer progression and development of chemotherapy resistances. This study was designed to investigate the involvement of retinoic acid receptor alpha (RARα) in RB progression and chemoresistance as well as the impact of miR-138, a potential RARα regulating miRNA. RARα and miR-138 expression in etoposide resistant RB cell lines and chemotherapy treated patient tumors compared to non-treated tumors was revealed by Real-Time PCR. Overexpression approaches were performed to analyze the effects of RARα on RB cell viability, apoptosis, proliferation and tumorigenesis. Besides, we addressed the effect of miR-138 overexpression on RB cell chemotherapy resistance. A binding between miR-138 and RARα was shown by dual luciferase reporter gene assay. The study presented revealed that RARα is downregulated in etoposide resistant RB cells, while miR-138 is endogenously upregulated. Opposing RARα and miR-138 expression levels were detectable in chemotherapy pre-treated compared to non-treated RB tumor specimen. Overexpression of RARα increases apoptosis levels and reduces tumor cell growth of aggressive etoposide resistant RB cells in vitro and in vivo. Overexpression of miR-138 in chemo-sensitive RB cell lines partly enhances cell viability after etoposide treatment. Our findings show that RARα acts as a tumor suppressor in retinoblastoma and is downregulated upon etoposide resistance in RB cells. Thus, RARα may contribute to the development and progression of RB chemo-resistance.