Expression Levels Of miR-21 Research Articles

THE MUCOSAL MICRORNA PROFILE RELATES TO AGE AND SEVERITY OF DISEASE IN PATIENTS WITH ULCERATIVE COLITIS

Despite significant evidence that the expression of several microRNA’s (miRNA) impacts on disease activity in patients with ulcerative colitis (UC), it remains unknown if the more severe disease phenotype seen in pediatric-onset UC can be explained by altered miRNA expression. In this study, we aimed to assess the relationship between miRNA expression, age, and disease severity in pediatric and adult patients with UC. Using RT-qPCR, we analyzed the expression of miR-21, miR-31, miR-126, miR-142, and miR-155 in paraffin-embedded rectum biopsies from 30 pediatric and 30 adult-onset UC patients, and found that lesions from adult patients had significantly higher expression levels of miR-21 compared to pediatric patients and that the expression levels of miR-31 (all patients) and miR-155 (pediatric patients only) correlated inversely with histological assessed disease severity. Using in situ hybridization followed by image analysis, the expression estimates of miR-21 and miR-126 were found to correlate with histological assessed disease severity. In conclusion, we found that the expression of miRNAs depends on the age of the patient and/or the severity of the disease, suggesting that miRNAs may contribute to the regulation of inflammation in UC and could be useful biomarkers in the surveillance of disease severity.

THE MUCOSAL MICRORNA PROFILE RELATES TO AGE AND SEVERITY OF DISEASE IN PATIENTS WITH ULCERATIVE COLITIS

Abstract Despite significant evidence that the expression of several microRNA’s (miRNA) impacts on disease activity in patients with ulcerative colitis (UC), it remains unknown if the more severe disease phenotype seen in pediatric-onset UC can be explained by altered miRNA expression. In this study, we aimed to assess the relationship between miRNA expression, age, and disease severity in pediatric and adult patients with UC. Using RT-qPCR, we analyzed the expression of miR-21, miR-31, miR-126, miR-142, and miR-155 in paraffin-embedded rectum biopsies from 30 pediatric and 30 adult-onset UC patients, and found that lesions from adult patients had significantly higher expression levels of miR-21 compared to pediatric patients and that the expression levels of miR-31 (all patients) and miR-155 (pediatric patients only) correlated inversely with histological assessed disease severity. Using in situ hybridization followed by image analysis, the expression estimates of miR-21 and miR-126 were found to correlate with histological assessed disease severity. In conclusion, we found that the expression of miRNAs depends on the age of the patient and/or the severity of the disease, suggesting that miRNAs may contribute to the regulation of inflammation in UC and could be useful biomarkers in the surveillance of disease severity.

Evaluation of miR-21 Expression in the Gastric Mucosa of Patients with Chronic Gastritis as the Predictive Factor of Gastric Cancer

The aim of the study was to evaluate the potential of miR-21 expression in the gastric mucosa as the marker of early stage gastric adenocarcinoma and cancer development. Material and methods . The study included the following materials: the first group consisted of surgical material of 60 stomachs (gastric cancer of intestinal type), in each case, 1 fragment of tumor tissue and 4 specimens of the gastric mucosa of the distant zone (2 – antrum mucosa and 2 – fundal mucosa) were taken; the second group consisted of biopsy material of 62 cases in patients with chronic gastritis (5 specimens according to the OLGA-system protocol). The level of miR-21 expression in tissues was determined by reverse transcription polymerase chain reaction. Results. miR-21 expression in samples with gastric cancer was significantly higher (median – 158, p=0.0006) compared with the group of endoscopic biopsies of the gastric mucosa (median – 42). The highest level of expression was observed in adenocarcinoma tissues (median – 270), while there were no significant differences when comparing it with the level of miR-21 expression in samples of the distant zone mucosa (p=0.2).тIncreased miR-21 expression may indicate the high risk group of developing gastric adenocarcinoma, probablyтbefore the appearance of histological signs of neoplasia. Conclusion. Regression analysis performed to confirm the predictive potential of miR-21 expression demonstrated 13-fold increased risk of adenocarcinoma development associated with an increase in miR-21 levels for every 50 units of relative expression normalized to small U6 RNA. The constructed statistical model, which includes morphological criteria for evaluating the gastric mucosa and miR-21 expression, may reflect the degree of violation of cellular epigenetic molecular pathways and allows increasing the prognostic value of a biopsy investigation and the accuracy of a stratified assessment of the gastric adenocarcinoma risk development

DNAzyme cascade circuits in highly integrated DNA nanomachines for sensitive microRNAs imaging in living cells

DNA molecular probes have emerged as powerful tools for fluorescence imaging of microRNAs (miRNAs) in living cells and thus elucidating functions and dynamics of miRNAs. In particular, the highly integrated DNA probes that can be able to address the robustness, sensitivity and consistency issues in a single assay system were highly desired but remained largely unsolved challenge. Herein, we reported for the first time that the development of the novel DNA nanomachines that split-DNAzyme motif was highly integrated in a single DNA triangular prism (DTP) reactor and can undergo target-activated DNAzyme catalytic cascade circuits, allowing amplified sensing and imaging of tumor-related microRNA-21 (miR-21) in living cells. The DNA nanomachines have shown dynamic responses for target miR-21 with excellent sensitivity and selectivity and demonstrated the potential for living cell imaging of miR-21. With the advantages of facile modular design and assembly, high biostability, low cytotoxicity and excellent cellular internalization, the highly integrated DNA nanomachines enabled accurate and effective monitoring of miR-21 expression levels in living cells. Therefore, our developed strategy may afford a reliable and robust nanoplatform for tumor diagnosis and for related biological research.

Transcriptomic analysis reveals essential microRNAs after peripheral nerve injury.

Studies have shown that microRNAs (miRNAs) mediate posttranscriptional regulation of target genes and participate in various physiological and pathological processes, including peripheral nerve injury. However, it is hard to select key miRNAs with essential biological functions among a large number of differentially expressed miRNAs. Previously, we collected injured sciatic nerve stumps at multiple time points after nerve crush injury, examined gene changes at different stages (acute, sub-acute, and post-acute), and obtained mRNA expression profiles. Here, we jointly analyzed mRNAs and miRNAs, and investigated upstream miRNAs of differentially expressed mRNAs using Ingenuity Pathway Analysis bioinformatic software. A total of 31, 42, 30, and 23 upstream miRNAs were identified at 1, 4, 7, and 14 days after rat sciatic nerve injury, respectively. Temporal expression patterns and biological involvement of commonly involved upstream miRNAs (miR-21, let-7, miR-223, miR-10b, miR-132, miR-15b, miR-127, miR-29a, miR-29b, and miR-9) were then determined at multiple time points. Expression levels of miR-21, miR-132, miR-29a, and miR-29b were robustly increased after sciatic nerve injury. Biological processes involving these miRNAs include multicellular organismal response to stress, positive regulation of the epidermal growth factor receptor signaling pathway, negative regulation of epithelial cell differentiation, and regulation of myocardial tissue growth. Moreover, we constructed mechanistic networks of let-7, miR-21, and miR-223, the most significantly involved upstream miRNAs. Our findings reveal that multiple upstream miRNAs (i.e., let-7, miR-21, and miR-223) were associated with gene expression changes in rat sciatic nerve stumps after nerve injury, and these miRNAs play an important role in peripheral nerve regeneration. This study was approved by the Experimental Animal Ethics Committee of Jiangsu Province of China (approval No. 20190303-18) on March 3, 2019.

MiR-21 Induces Chemoresistance in Ovarian Cancer Cells via Mediating the Expression and Interaction of CD44v6 and P-gp.

BackgroundOvarian cancer (OC), a representative female reproductive system tumor, is one of the most malignant tumors in female. The most important reason for its poor prognosis is because of its high rate of chemotherapy resistance.ResultsThis study aims to explore the effects of miR-21 on the chemotherapy resistance of OC cells. The functions of miR-21 on proliferation, migration and invasion of OC cells were assessed by transwell, clonal formation and CCK8 assay. Expression levels of miR-21, P-gp and CD44v6 in SKOV3 (cisplatin sensitive) cells and SKOV3/DDP (cisplatin resistant) cells were detected by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting. Si-CD44v6 was transfected into OC cells to detect the influence on P-glycoprotein (P-gp) expression. Immunofluorescence was used to detect the localization of CD44v6 and P-gp in cell. Co-immunoprecipitation was used to detect the relationship between CD44v6 and P-gp. Results showed that miR-21 expression in cisplatin-resistant SKOV3/DDP cells was significantly higher than that in SKOV3 cells, at the same time, cells proliferation, as well as invasion and migration ability were enhanced after the miR-21 mimics transfected into SKOV3 cisplatin-sensitive cells. Furthermore, miR-21 expression level affected the CD44v6 and P-gp expression. Immunofluorescence and co-immunoprecipitation showed that CD44v6 and P-gp protein could interact.ConclusionIn conclusion, the high miR-21 expression level could increase the proliferation, invasion, and migration ability of OC cells. And the interaction of CD44v6 and P-gp may mediate miR-21 involvement in chemotherapy resistance of OC cells.

Investigation of the efficacy of paclitaxel on some miRNAs profiles in breast cancer stem cells.

Understanding of the functions of microRNAs in breast cancer and breast cancer stem cells have been a hope for the development of new molecular targeted therapies. Here, it is aimed to investigate the differences in the expression levels of let-7a, miR-10b, miR-21, miR-125b, miR-145, miR-155, miR-200c, miR-221, miR-222 and miR-335, which associated with gene and proteins in MCF-7 (parental) and MCF-7s (Mammosphere/stem cell-enriched population/CD44+/CD24-cells) cells treated with paclitaxel. MCF-7s were obtained from parental MCF-7 cells. Cytotoxic activity of paclitaxel was determined by ATP assay. Total RNA isolation and cDNA conversion were performed from the samples. Changes in expression levels of miRNAs were examined by RT-qPCR. Identified target genes and proteins of miRNAs were analyzed with RT-qPCR and western blot analysis, respectively. miR-125b was significantly expressed (2.0946-fold; p = 0.021) in MCF-7s cells compared to control after treatment with paclitaxel. Downregulation of SMO, STAT3, NANOG, OCT4, SOX2, ERBB2 and ERBB3 and upregulation of TP53 genes were significant after 48 h treatment in MCF-7s cells. Protein expressions of SOX2, OCT4, SMAD4, SOX2 and OCT4 also decreased. Paclitaxel induces miR-125b expression in MCF-7s cells. Upregulation of miR-125b may be used as a biomarker for the prediction of response to paclitaxel treatment in breast cancer.

Role of circulating micro-RNA-21 as a noninvasive biomarker for detection of colorectal cancer

ObjectiveTo assess serum micro-RNA-21 (miR-21) expression level as a stable blood-based biomarker for colorectal cancer (CRC).BackgroundCRC represents the third most common tumor worldwide with male predominance, with significant morbidity and mortality despite the recent therapeutic improvements. Therefore, a noninvasive diagnostic marker with good sensitivity and specificity is very important, which leads to better clinical outcomes. MiR-21 is an oncogenic miRNA that modulates the expression of multiple cancer-related target genes. MiR-21 expression was upregulated in the serum and in the malignant tissues in relation to the normal tissues. Its elevated level is claimed to be associated with poor survival and poor response to treatment.Patients and methodsThis cross-sectional study was carried on 50 patients and measured their serum miR-21 (CYBR Green). The cases group comprised 20 consecutive patients with CRC proved by colonoscopy and histopathological biopsy examination, the second group comprised 20 patients with colorectal polyps with no malignant transformation proved by histopathological examination, and the third group included 10 patients with normal colonoscopy finding as the control group. The studied cases were attending the Endoscopy Unit Menoufia University Hospital and Nasser Institute. The three groups agreed to written consent to participate in this study after the approval of the local ethical committee.ResultsSerum miR-21 expression was significantly upregulated in the cases group of patients with CRC compared with the other groups, with P value less than 0.001, showing sensitivity of 85% and specificity of 83.3% at a cutoff point of −2.31.ConclusionMi-RNA-21 shows upregulation in the serum in CRC cases, with statistically significant difference compared with the control group and the polyp group, showing promising results as a noninvasive biomarker of CRC.

Prognostic Values of MicroRNA-21 and Ki-67 in Diffuse Large B-Cell Lymphoma Patients: Egyptian Experience.

MicroRNA-21 (miR-21) is a small non-coding RNA which influences tumorigenesis by inhibiting the expression of target genes. Ki-67 is a nucleolar antigen highly correlated with the rate of proliferating cells. In this study, we aimed to evaluate the prognostic impact of miR-21 and Ki-67 in DLBCL disease in a cohort of Egyptian patients. We prospectively enrolled 53 newly diagnosed DLBCL patients. RT-PCR was used to evaluate the plasma expression levels of miR-21. Tissue Ki-67 was assessed using immunohistochemistry (IHC) of lymph node biopsy sections. Overall survival (OS) and progression free survival (PFS) were the primary outcomes. miR-21 expression was significantly higher in patients with DLBCL in comparison to controls (p < 0.001). The median Ki-67 expression was 70% and positivity ranged from 25% to 100%. Response to treatment was achieved in 23 patients (43.4%). Higher miR-21 was associated with poor response to treatment (p = 0.03). Although patients' age was a significant predictor of OS in univariate analysis, none of the studied factors could predict OS in multivariate analysis. However, we found that Ki-67 expression was a significant predictor of PFS in both univariate and multivariate analyses. The study suggested that plasma miR-21 might be a valuable non-invasive prognostic marker of response to treatment in DLBCL patients. Moreover, Ki-67 is a potential significant predictor of both OS and PFS in those patients.

Identification of Serum microRNA-25 as a novel biomarker for pancreatic cancer.

To identify serum microRNA-25 (miR-25) as a diagnostic biomarker for pancreatic cancer (PCa) and to evaluate its supplementary role with serum carbohydrate antigen 19-9 (CA19-9) in early identification of cancers.Eighty patients with pancreatic cancer and 91 non-cancer controls were enrolled in this study. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression level of miR-25. Levels of CA19-9, carcinoembryonic antigen (CEA) and carbohydrate antigen 125 (CA125) were measured by chemiluminescent immunoassay. The logistic model was established to evaluate the correlation of miR-25 with clinical characteristics. A risk model for PCa was conducted by R statistical software. Diagnostic utility for PCa and correlation with clinical characteristics were analyzed.The expression level of miR-25, in the PCa group was significantly higher (P < .05). Risk Model illustrated the relation between miR-25 and pancreatic cancer. With the combination of CA19-9, the performance of miR-25 in early stages (I+II) in the diagnosis of PCa was profoundly better than CA19-9 and miR-25 alone. This combination was more effective for discriminating PCa from non-cancer controls (AUC-ROC, 0.985; sensitivity, 97.50%; specificity, 90.11%) compared with CA19-9 alone or the combination of CA19-9 and CA125.The expression level of miR-25 among pancreatic cancer patients was significantly higher than that in the control group. miR-25 existed as one of the most relevant factors of PCa. miR-25 can serve as a novel noninvasive approach for PCa diagnosis, and with the supplementary of CA19-9, the combination was more effective, especially in early tumor screening.

The Prognostic Significance of miR-21 Expression among Surgically Resected Hepatocellular Carcinoma Patients: Evidence from a Meta-Analysis and Retrospective Cohort Study.

Background To date, microRNA-21 (miR-21) has been reported to be associated with the prognosis of hepatocellular carcinoma (HCC) in various studies, yet the results were inconsistent. The purpose of this two-part study, consisting of a retrospective cohort study and a meta-analysis, sets out to determine the prognostic role of miR-21 expression among HCC patients who underwent surgical resection. Methods. In this study, we first detected miR-21 expression in HCC patients by quantitative real-time PCR (qRT-PCR). Patients were divided into a high miR-21 expression group and a low miR-21 expression group according to the median level of miR-21 expression in tumor tissues. The survival outcomes of the two groups were analyzed by the Kaplan-Meier method with the log-rank test. Multivariate analysis of the prognostic factors was performed with the Cox regression model. Subsequently, eligible studies were obtained by searching on PubMed, Cochrane Library, and Web of Science, and a meta-analysis was performed to assess the prognostic role of miR-21 expression among HCC. Results The qRT-PCR analysis results of our cohort study showed miR-21 expression was significantly upregulated in HCC tissues when compared with adjacent nontumor tissues. Multivariate analysis suggested that miR-21 expression was an independent prognostic factor for overall survival (OS) (hazard ratio, HR = 2.361) and disease-free survival (DFS) (HR = 2.024) in HCC patients who underwent surgical resection. A total of 10 studies with 969 patients were enrolled in the meta-analysis, consisting of 9 studies from the database search and our cohort study. We observed that elevated miR-21 expression can predict poor OS (HR = 2.24, 95%CI = 1.73‐2.91, P < 0.001) and DFS/recurrence-free survival (RFS) (HR = 2.44, 95%CI = 1.62‐3.67, P < 0.001) in surgically resected HCC patients. Conclusions Our study demonstrated that miR-21 high expression among surgically resected HCC patients is a prognostic factor that indicated adverse survival.

SDF-1α/MicroRNA-134 Axis Regulates Nonfunctioning Pituitary Neuroendocrine Tumor Growth via Targeting VEGFA.

BackgroundNonfunctioning pituitary neuroendocrine tumor (NF-PitNET) is difficult to resect. Except for surgery, there is no effective treatment for NF-PitNET. MicroRNA-134 (miR-134) has been reported to inhibit proliferation and invasion ability of tumor cells. Herein, the mechanism underlying the effect of miR-134 on alleviating NF-PitNET tumor cells growth is explored.MethodsMouse pituitary αT3-1 cells were transfected with miR-134 mimics and inhibitor, followed by treatment with stromal cell-derived factor-1α (SDF-1α) in vitro. MiR-134 expression level: we used quantitative real-time PCR (qRT-PCR) to detect the expression of miR-134. Cell behavior level: cell viability and invasion ability were assessed using a cell counting kit-8 (CCK8) assay and Transwell invasion assay respectively. Cytomolecular level: tumor cell proliferation was evaluated by Ki-67 staining; propidium iodide (PI) staining analyzed the effect of miR-134 on cell cycle arrest; western blot analysis and immunofluorescence staining evaluated tumor migration and invasive ability. Additionally, we collected 27 NF-PitNET tumor specimens and related clinical data. The specimens were subjected to qRT-PCR to obtain the relative miR-134 expression level of each specimen; linear regression analysis was used to analyze the miR-134 expression level in tumor specimens and the age of the NF-PitNET population, gender, tumor invasion, prognosis, and other indicators.Results In vitro experiment, miR-134 was observed to significantly inhibit αT3-1 cells proliferation characterized by inhibited cell viability and expressions of vascular endothelial growth factor A (VEGFA) and cell cycle transition from G1 to S phase (P < 0.01). VEGFA was verified as a target of miR-134. Additionally, miR-134-induced inhibition of αT3-1 cell proliferation and invasion was attenuated by SDF-1α and VEGFA overexpression (P < 0.01). In primary NF-PitNET tumor analysis, miR-134 expression level was negatively correlated with tumor invasion (P = 0.003).ConclusionThe regulation of the SDF-1α/miR-134/VEGFA axis represents a novel mechanism in the pathogenesis of NF-PitNETs and may serve as a potential therapeutic target for the treatment of NF-PitNETs.

Inhibition of miR-152 during In Vitro Maturation Enhances the Developmental Potential of Porcine Embryos.

Simple SummaryMiR-152 is a highly conserved miRNA across different species and plays a role in the regulation of cell differentiation, proliferation, and survival. However, the exact role of miR-152 in oocyte and embryo development is not yet known. In this study, we specifically manipulated the expression level of miR-152 in porcine cumulus-oocyte complexes (COCs) and monitored their developmental competence until the blastocyst stage. We mainly found that a suppressed expression of miR-152 during oocyte maturation significantly improved the blastocyst rate. Our results indicate that this negative correlation between miR-152 during oocyte maturation and the blastocyst rate in pigs could be through targeting IGF system components during oocyte development. These results provide more insights into the role of miRNAs during oocyte and embryonic development that could improve the in vitro production system for mammalian embryos. Oocyte developmental competence is regulated by various mechanisms and molecules including microRNAs (miRNAs). However, the functions of many of these miRNAs in oocyte and embryo development are still unclear. In this study, we managed to manipulate the expression level of miR-152 during oocyte maturation to figure out its potential role in determining the developmental competence of porcine oocytes. The inhibition (Inh) of miR-152 during oocyte maturation does not affect the MII and cleavage rates, however it significantly enhances the blastocyst rate compared to the overexpression (OvExp) and control groups. Pathway analysis identified several signaling pathways (including PI3K/AKT, TGFβ, Hippo, FoxO, and Wnt signaling) that are enriched in the predicted target genes of miR-152. Gene expression analysis revealed that IGF1 was significantly up-regulated in the Inh group and downregulated in the OvExp group of oocytes. Moreover, IGF1R was significantly upregulated in the Inh oocyte group compared to the control one and IGFBP6 was downregulated in the Inh oocyte group compared to the other groups. Blastocysts developed from the OvExp oocytes exhibited an increase in miR-152 expression, dysregulation in some quality-related genes, and the lowest rate of blastocyst formation. In conclusion, our results demonstrate a negative correlation between miR-152 expression level and blastocyst rate in pigs. This correlation could be through targeting IGF system components during oocyte development.

Caveolin-1 regulates the expression of miR-183 and inhibits the invasion and migration of invasive pituitary adenomas by affecting early growth response 1 (EGR1)/Krueppel-like factor 5 (KLF5) interaction

This study aims to investigate the mechanism through which Caveolin-1 (CAV-1) regulates the expression of micro ribonucleic acid (miR)-183 in invasive pituitary adenoma (IPA) tissues and GH3 cells, and explore the effects of CAV-1 and miR-183 on the invasion and migration ability of GH3 cells. Western blotting was used to detect the expression level of CAV-1, early growth response 1 (EGR1) and Krueppel-like factor 5 (KLF5). Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect the expression of miR-183. The mechanisms of interaction between CAV-1, EGR1, and KLF5 were studied by immunoprecipitation experiments. Transwell and cell scratch tests were used to determine the invasion and migration ability of GH3 cells. The dual-luciferase reporter gene experiment was used to detect the effects of EGR1 and KLF5 on miR-183 luciferase activity and verify the targeting relationship between miR-183 and ezrin. The expression of CAV-1 was up-regulated. However, following the knockdown of CAV-1, the invasion and migration ability of GH3 cells was significantly inhibited (P<0.05). The expression of miR-183 was down-regulated, but the expression level of miR-183 was markedly increased following the knockdown of CAV-1 (P<0.05). The knockdown of CAV-1 inhibited the nuclear ectopic of the EGR1 protein in GH3 cells. At the same time, the interaction between EGR1 and KLF5 in GH3 cells was significantly inhibited (P<0.05). The luciferase activity of miR-183 increased significantly after overexpression of KLF5 while overexpression of EGR1 and KLF5 had no significant effect on intracellular luciferase activity. Overexpression of miR-183 markedly inhibited the luciferase activity of wild-type EZR and the expression of the EZR protein in GH3 cells. Furthermore, the overexpression of miR-183 or the inhibition of EZR can reduce the invasion and migration ability of GH3 cells. The simultaneous overexpression or inhibition of miR-183 and EZR expression has no obvious effect on the invasion and migration ability of GH3 cells. CAV-1 up-regulates the expression of miR-183 by inhibiting the nuclear ectopic of EGR1 and the interaction between EGR1 and KLF5 in GH3 cells. Also, miR-183 negatively regulates the expression of EZR and inhibits the invasion and migration of GH3 cells.

Differential Expression of miR-21, miR-23a, and miR-27a, and Their Diagnostic Significance in Egyptian Colorectal Cancer Patients.

Background: Colorectal cancer (CRC) rates are affected by genetics, ethnicity, and environmental factors; it is considered one of the most aggressive human malignancies with high mortality and morbidity rates worldwide due, in part, to its asymptomatic nature during the early stages of disease. Objective: Owing to the impact of microRNA (miRNA) dysregulation on CRC development and progression, this study was conducted to explore the expression levels of mir-21, -23a, and -27a in the sera and tissues of Egyptian CRC patients and to evaluate their diagnostic efficacy based on circulating levels. Methods: In the test phase, the relative expression levels of the studied miRNAs were evaluated in the sera of 70 participants (35 CRC patients and 35 healthy controls) using quantitative real-time-polymerase chain reaction and to verify their diagnostic value. The exploratory phase was designed to validate the tumor-derived trait by comparing the miRNA levels in the cancerous and adjacent noncancerous tissues. Results: The relative expression levels of the studied miRNAs were significantly upregulated in both serum and tumor tissues of the patients compared to their corresponding controls. In addition, significant positive correlations were found between the relative expression levels of the studied miRNAs in serum samples and their levels in the matched CRC tissues. The serum expression levels of mir-21 and -23a were more predictive of CRC than mir-27a. Conclusion: Circulating mir-21, -23a, and -27a expression levels appear to be valuable diagnostic biomarkers for CRC, especially when combined.

Evaluation of Serum miR-21 Expression Level in Patients Suspected of Mycobacterium tuberculosis

Evaluation of Serum miR-21 Expression Level in Patients Suspected of Mycobacterium tuberculosis

Modulatory response of chrysin supplementation in a experimental autoimmune encephalomyelitis model: Evaluation of microRNAs influence

BackgroundMultiple sclerosis (MS) is a chronic autoimmune disease which causing demyelination, resulting in several damages to central nervous system. NonCoding microRNAs (miRNAs) are post transcriptional gene expression regulators that have been associated with MS-pathophysiology. In this context, chrysin is a bioactive compound that has several pharmacological, nutraceutical and neuroprotective effects. Thus, the aim of study was to investigate the effect of chrysin supplementation in alterations caused by experimental autoimmune encephalomyelitis (EAE) model in mice, analyzing histological parameters and changes in expression levels of MIR21, MIR155 and MIR326 (miRNAs-21, 155 and 326). MethodsFor induction of EAE was used myelin oligodendrocyte glycoprotein (35–55) peptide in C57BL/6 mice. EAE model caused increase of inflammatory infiltrate in SC and upregulated expression levels of miRNAs analyzed. ResultsSupplementation with chrysin was able to decrease of miRNAs-21 and 155 expression levels altered in EAE. Chrysin supplementation did not modulate miRNA-326 expression levels. ConclusionsThe findings suggest that chrysin attenuated inflammatory process presented in histological analysis, probably due to its indirect modulatory effect on miRNAs-21 and 155. Therefore, chrysin supplementation may represent an advance for MS therapeutic, however, further studies are needed to understand the role of miRNAs-21 and 155 in your modulatory response.

Study on the clinical significance of miR-100 expression in the invasion and metastasis of hepatocellular carcinoma

Objective: To study the miR-100 expression levels in the tissues of hepatocellular carcinoma patients, and to further explore the correlation between miR-100 and the invasion and metastasis of hepatocellular carcinoma cells and its effect on patients' prognostic survival. Methods: Clinicopathological data of 70 cases that underwent hepatectomy from December 2013 to December 2016 in the Department of Hepatobiliary and Pancreatic Surgery of Henan Provincial People's Hospital were retrospectively analyzed. Real-time fluorescent quantitative PCR was used to detect the different miR-100 expression levels in cancerous and adjacent tissues. The expression of miR-100 with different clinicopathological features was compared, and the prognostic factors of patients with hepatocellular carcinoma were comprehensively analyzed. The correlation between miR-100 and patients' clinicopathological features was tested by χ(2). Kaplan-Meier method was used to draw the survival curve. Log-rank test was used to examine the survival rate difference in each subgroup. Cox regression model was used to analyze the multivariate prognosis. Results: miR-100 expression was down-regulated to a different degree in hepatocellular carcinoma tissues than the corresponding adjacent tissues. Among them, the down-regulated expression of miR-100 in hepatocellular carcinoma tissues accounted for 82.9% (58/70, P < 0.05) of all cases when compared to corresponding paracancerous tissues. miR-100 expression level was significantly correlated to high Edmondson's grade, high TNM stage and intrahepatic metastasis (P < 0.05). The overall survival time of miR-100 positive expression was significantly higher than that of miR-100 negative expression (Log-rank χ(2) = 8.257, P < 0.05). Univariate survival analysis results revealed that the miR-100 expression level, tumor size, TNM stage, Edmondson's grade, and presence or absence of venous tumor thrombosis had a poor prognosis (P < 0.05). Cox multivariate regression analysis showed that the tumor size, Edmondson's grade, and miR-100 expression level were independent factors affecting the prognostic survival in hepatocellular carcinoma patients. In addition, patients with low positive expression rate of miR-100, large tumors and high Edmondson's grade had a poor prognosis. Conclusion: The level of miR-100 expression in hepatocellular carcinoma cells is low, so it is closely related to the invasion and metastasis and affects the prognostic survival of hepatocellular carcinoma patients.

Evaluation of color Doppler ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of breast cancer.

To evaluate the clinical value of color Doppler ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of breast cancer (BC). 129 patients with BC and 50 patients with benign breast lesions were selected. They were scanned by color Doppler ultrasound on the breast. The relative expression of miR-21 and miR-27a in plasma was detected by real-time fluorescence quantitative PCR. The clinical value of color Doppler ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of BC was analyzed retrospectively. Two-dimensional ultrasound images of BC showed irregular shape, unclear boundary, burr-like edge, uneven internal echo, weakened rear echo, microcalcification, aspect ratio > 1 and axillary lymph node metastasis. The grade of blood flow signal was improved, mainly in grade II and above, which was statistically different from that of benign control group (P < 0.001). The relative expression of plasma miR-21 and miR-27a in breast cancer group was significantly higher than that in benign control group (P < 0.001). The relative expressions of plasma miR-21 and miR-27a in breast cancer group were significantly correlated with histological grade, clinical stage, lymph node metastasis and postoperation. The relative expression levels of miR-21 and miR-27a in plasma of patients with low histological grade (grade III), high clinical stage (stage III, IV) and lymph node metastasis before operation were significantly higher than those in patients with high histological grade (grade I, II), low clinical stage (stage I, II) and no lymph node metastasis after operation (P < 0.01). The sensitivity, accuracy and negative predictive value of ultrasound combined with plasma miR-21 and miR-27a in the diagnosis of BC were significantly higher than those of each single examination. Color Doppler ultrasound combined with plasma miR-21 and miR-27a can significantly improve the diagnostic efficiency of BC, which is beneficial to early diagnosis and early clinical intervention.

Arecoline enhances miR-21 to promote buccal mucosal fibroblasts activation

Oral submucous fibrosis (OSF) is an irreversible fibrosis disease and a potentially malignant disorder in the oral cavity. Various studies have shown that miR-21 was implicated in the fibrogenesis and carcinogenesis, but its functional role in the development of OSF has not been investigated. The expression levels of miR-21 in arecoline-stimulated normal buccal mucosal fibroblasts (BMFs) and OSF specimens were determined by qRT-PCR. Exogenous administration of TGF-β and its inhibitor (SB431542) were utilized to examine the involvement of TGF-β signaling in miR-21 alteration. Collagen gel contraction, transwell migration, and invasion assays were used to assess the myofibroblast activities. The relationship between α-SMA and miR-21 was calculated using the Pearson correlation coefficient. MiR-21 expression was induced in BMFs by arecoline treatment in a dose-dependent manner. Our results showed that this upregulation was mediated by TGF-β signaling. Subsequently, we demonstrated that the administration of the miR-21 inhibitor suppressed the arecoline-induced myofibroblast characteristics, including a higher collagen gel contractility and cell motility, in normal BMFs. Furthermore, inhibition of miR-21 was sufficient to attenuate the myofibroblast features in fibrotic BMFs. Besides, we showed that the expression of miR-21 was aberrantly upregulated in the OSF tissues and there was a positive correlation between miR-21 and myofibroblast marker, α-SMA. MiR-21 overexpression in OSF may be due to the stimulation of areca nut, which was mediated by the TGF-β pathway. Our data suggested that the repression of miR-21 was a promising direction to palliate the development and progression of OSF.