Serum miR-21 Research Articles

The expression of miR-155 and miR-181 in gastric cancer and their effects on clinical parameters and prognostic value*

Abstract Objective The aim of this study was to investigate the expression and clinical significance of miR-155 and miR-181 in gastric cancer. Methods Sixty-eight patients with gastric cancer and 80 healthy volunteers were selected as subjects. Serum samples of patients and volunteers were collected to detect miR-155 and miR-181 expression levels in serum and tumor tissues. Results miR-155 and miR-181 serum levels were significantly higher in patients with gastric cancer than those in the control group (P < 0.05). miR-155 and miR-181 expression levels in gastric cancer tissues were 16.74 ± 4.29 and 12.17 ± 3.26, respectively and 3.42 ± 0.39) and 3.06 ± 0.69 in paracancerous tissues, respectively, and they were 1.22 ± 0.21 and 1.08 ± 0.35 in normal tissues, respectively. miR-155 and miR-181 expression levels in cancer tissues were significantly higher than adjacent tissues, and they were lowest in normal tissues (P < 0.05). High miR-155 and miR-181 expression had significant effects on differentiation, T stage, lymph node metastasis, and clinical stage (P < 0.05). Cox multivariate analysis showed that onset, T stage, clinical stage, differentiation degree, lymph node metastasis, miR-155, and miR-181 were independent risk factors for prognosis of patients with gastric cancer. Conclusion Serum miR-155 and miR-181 have a diagnostic value in gastric cancer, and they are strong signals of poor prognosis.

Association of circulating miR-20a, miR-27a, and miR-126 with non-alcoholic fatty liver disease in general population

Non-alcoholic fatty liver disease (NAFLD) is closely associated with obesity, metabolic syndrome, and type II diabetes mellitus. Recently, circulating microRNAs (miRNAs) have been proposed as useful disease biomarkers. We examined whether circulating miRNAs, such as miR-20a, miR-27a, and miR-126, were useful biomarkers for NAFLD. We conducted a cross-sectional analysis of 527 subjects aged 39 years or older who had undergone a health examination in the Yakumo Study. Of the residents, 92 were diagnosed with NAFLD using a registered medical sonographer. Serum miR-20a, miR-27a and miR-126 levels were measured by quantitative real-time PCR. We then calculated the odds ratios for serum miRNA level changes according to the severity of NAFLD using normal liver status as the reference group. Serum levels of miR-20a and 27a, but not miR-126, were significantly lower in NAFLD subjects than normal subjects. Serum miR-20a and miR-27a levels were significantly lower in both male and female severe NAFLD subjects. Logistic regression analysis showed a significant relationship between low circulating miR-20a and 27a levels and severe NAFLD. Down-regulated circulating miR-20a and 27a levels were significantly associated with severe NAFLD in the general population. Circulating miR-20a and miR-27a may be useful biomarkers for severe NAFLD.

The expression level of microRNA-122 in patients with type 2 diabetes mellitus in correlation with risk and severity of coronary artery disease

BackgroundType 2 diabetes mellitus (T2DM) has reached epidemic proportions worldwide. Coronary artery disease (CAD) is one of the most important causes of mortality worldwide. MicroRNAs (miRNAs) modulate gene expression and is involved in the pathogenesis of T2DM and CAD. The objective of the current study was to explore the expression pattern of miR-122 in T2DM with or without CAD. Moreover, we aimed to evaluate the association between miR-122 and risk and severity of CAD in T2DM.Participants and methodsThis cross-sectional controlled study enrolled 130 patients with T2DM and 110 control group. The enrolled diabetic patients were classified into two groups: seventy patients without CAD and 60 patients without CAD. All patients were investigated using a 12-lead standard ECG, echocardiography, and coronary arteriography. The serum MiR-122 expression profile was measured using quantitative real-time (qRT) PCR.ResultsmiRNA-122 expression levels were significantly higher in T2DM, in particular patients with T2DM with CAD, compared with the control group. Interestingly, miRNA-122 expression levels were positively correlated with cardiometabolic risks and severity of coronary occlusion. Linear regression analysis test showed that miRNA-122 were independently correlated with high-density lipoprotein, ejection fraction, and uric acid. The power of miRNA-122 expression level to diagnose T2DM among studied participants was evaluated using receiver operating characteristic. The area under curve was 0.997 (95% confidence interval=0.993–1.00), with sensitivity of 96.9% and specificity of 99%, and regarding the power for differentiating patients with T2DM with CAD from patients with T2DM without CAD, the area under curve was 0.832 (95% confidence interval=0.763–0.902), with sensitivity of 93.3% and specificity of 96.86%.ConclusionThe miRNA-122 expression levels were higher in the T2DM group compared with controls, in particular patients with CAD. The higher levels of miR-122 expression were strongly correlated with cardiometabolic risk factors and severity of coronary occlusion. Therefore, miR-122 expression levels seem to be a noninvasive biomarker for CAD.

Circulating microRNAs (miR-21, miR-223, miR-885-5p) along the clinical spectrum of HCV-related chronic liver disease in Egyptian patients

Background and study aimsMicroRNAs (miRNAs), small single stranded RNAs, function in the post-transcriptional regulation of gene expression and incorporated in pathogenesis of HCV related chronic liver disease. This study was designed to evaluate the significance of serum miR-21, miR-223, and miR-885-5p as biomarkers in various clinicopathological stages of HCV related chronic liver disease. Patients and methodsSerum miR-21, miR-223, and miR-885-5p were quantified by quantitative RT PCR in 60 patients with HCV-related liver disease (presumably genotype 4), in addition to 25 healthy controls. HCV patients were classified into: chronic non-cirrhotic HCV (n = 15), HCV related liver cirrhosis (n = 15), and hepatocellular carcinoma (HCC) (n = 30). Results: Serum levels of miR-885-5p in cirrhotic patients ± HCC (n = 45) were significantly higher than the non-cirrhotic patients (n = 15); p = 0.007 and healthy control; p = 0.001. However, no such significance was detected between HCC and non-HCC HCV patients; p = 0.12. Serum miRNA-885-5p was able to discriminate cirrhosis ± HCC from healthy controls using ROC analysis; AUC 0.85, 87% sensitivity and 80% specificity. On the other hand, HCC patients had significantly higher serum miR-2 1evels than non-HCC patients (non-cirrhotic and cirrhotic groups, n = 30); p = 0.048 and the control group; p = 0.002. ROC could differentiate HCC from control group; AUC 0.89, 80% sensitivity, 80% specificity. Both serum bilirubin and albumin showed significant weak correlation with miRNA-885-5p (r = 0.42, p = 0.001) and (r = −0.27, p = 0.04), respectively but no such correlation was observed with serum miRNA-21. In contrast, miRNA-223 showed no significant difference across the studied groups. ConclusionAlong the spectrum of HCV-related chronic liver disease, miR-885-5p could be a potential marker for advanced liver damage while miR-21 could be a helpful diagnostic marker for HCC.

Serum microRNA Prognostic Model and Underlying Immune Alterations in Diffuse Large B Cell Lymphoma

Serum microRNA prognostic model and underlying immune alterations in diffuse large B cell lymphoma Abstract Background: MicroRNAs (miRs) have been investigated as prognostic biomarkers and therapeutic targets in diffuse large B-cell lymphoma (DLBCL). Translating these findings into clinical approaches should be validated by independent patient cohorts and a better understanding of underlying biology mechanism remains great interests. Methods: MiRNA microarray was performed on serum samples of 20 DLBCL patients at diagnosis, remission and at relapse. Subsequent real-time PCR validation of 504 newly diagnosed DLBCL patients was used to generate a serum prognostic model in predicting clinical outcomes. The mechanism of action of miR signature on tumor progression was further analyzed by whole exome sequencing and RNA sequencing. Results: Four serum miRs (miR-21, miR-155, miR-130b and miR-28) were identified to predict relapse in a training cohort of 225 DLBCL patients and validated in a validation cohort of 279 DLBCL patients. Multivariate Cox regression models indicated that when the International Prognostic Index was controlled, this serum miR prognostic model was an independent prognostic factor in DLBCL. Moreover, mutations of transcription factors (TFs) were significantly correlated with these four miRs. Twenty lymphoma-associated TFs regulated at least one miR by transmiR v2.0 database and different genetic mutation burden was present between low-index and high-index of the model. Furthermore, RNA sequencing data manifested that the four-miR signature correlated with immune status of the DLBCL patients. Conclusions: An easy-to-use miR prognostic model was utilized to predict disease relapse and prognosis of DLBCL patients. The tumor-specific mutation burden and immune dysregulation potentially contribute to tumor progression in DLBCL. Key words: MicroRNAs, Diffuse Large B-cell Lymphoma, Prognosis, Signaling pathways. Disclosures No relevant conflicts of interest to declare.

Low serum miR-223 expression predicts poor outcome in patients with acute myeloid leukemia.

BackgroundIdentification of biomarkers for acute myeloid leukemia (AML) is important for treating this malignancy. Recent studies have reported that microRNAs (miRNAs) are stably detectable in the blood/plasma and can be used as biomarkers for various types of cancer including AML. The aim of this study was to analyze miR‐223 level in serum as a potential indicator for AML diagnosis and prognosis prediction.MethodsQuantitative reverse transcription‐polymerase chain reaction (qRT‐PCR) was used to detect the levels of miR‐223 in the serum samples from 131 patients and 70 healthy individuals.ResultsThe results revealed that serum miR‐223 was underexpressed in AML patients, particularly those in intermediate and unfavorable cytogenetic risk groups. Further analysis revealed that serum miR‐223 could yield a receiver operating characteristic (ROC) area under the curve (AUC) of 0.849 with 83.2% sensitivity and 81.4% specificity. Moreover, a significant increase in serum miR‐223 level was observed in AML subjects after their treatment. Reduced serum miR‐223 level was highly associated with aggressive clinical variables and shorter survival of patients. Furthermore, miR‐223 expression was identified to be an independent prognostic predictor of worse overall survival.ConclusionIn conclusion, miR‐223 may be a reliable diagnostic and prognostic biomarker for AML.

Clinical value of LHPP-associated microRNAs combined with protein induced by vitamin K deficiency or antagonist-II in the diagnosis of alpha-fetoprotein-negative hepatocellular carcinoma.

BackgroundAlpha‐fetoprotein (AFP) has received extensive attention in the differential diagnosis of hepatocellular carcinoma (HCC), especially for AFP‐negative HCC (AFP‐NHCC). The current study aimed to explore the value of targeted regulation of LHPP expression‐related microRNAs (miRs) and protein induced by vitamin K deficiency or antagonist‐II (PIVKA‐II) in the differential diagnosis of AFP‐NHCC.MethodsA retrospective study was conducted on a testing set—including 214 AFP‐NHCC patients, 200 cirrhosis, and 210 controls, and a validation set—including 140 AFP‐NHCC patients, 134 cirrhosis, and 128 controls recruited from The First Affiliated Hospital of Hunan Normal University. Serum miRs were examined using quantitative real‐time PCR method. Serum PIVKA‐II was measured by enzyme‐linked immunosorbent assay.ResultsCompared with adjacent tissues, LHPP protein levels in cancer tissues were significantly decreased (P < .05). Predictive software and dual‐luciferase reporter assays showed that miR‐363‐5p and miR‐765 can target LHPP expression. Serum miR‐363‐5p, miR‐765, and PIVKA‐II levels were significantly higher in AFP‐HCC patients than in cirrhosis and controls. A logistic regression model combining miR‐363‐5p, miR‐765, and PIVKA‐II was performed. This model presented a high discriminating value (AUC: 0.930, sensitivity/specificity: 79.4%/95.4%) than any single indicator. In the validation set, this model still showed a high discriminating value (AUC: 0.936, sensitivity/specificity: 83.6%/94.7%).ConclusionCurrent model combining serum miR‐363‐5p, miR‐765, and PIVKA‐II has potential significance for diagnosis of AFP‐NHCC.

Circulating MicroRNA-122 and Fibrosis Stage Predict Mortality of Japanese Patients With Histopathologically Confirmed NAFLD.

The impact of circulating microRNA‐122 (miR‐122) on mortality in patients with histopathologically confirmed nonalcoholic fatty liver disease (NAFLD) remains unclear. We analyzed the overall survival rates in 441 Japanese patients with histopathologically confirmed NAFLD after a median follow‐up period of 4.7 years. We also determined the clinicopathologic, genetic, and epigenetic parameters, including serum miR‐122 levels, for prediction of mortality. Of the 441 study patients, 21 (4.8%) died during the follow‐up period. The cumulative survival rates were 95.4% and 90.6% at the end of 5 and 10 years, respectively. Multivariate analysis identified history of liver cancer (presence; hazard ratio [HR], 4.94; 95% confidence interval [CI], 1.84‐13.3), serum miR‐122 (<1.00 fold change; HR, 4.35; 95% CI, 0.06‐0.83), and fibrosis‐4 index (FIB‐4 index) (≥1.30; HR, 15.7; 95% CI, 2.01‐122) as significant risk factors of mortality. Cumulative survival rates varied significantly among patients with none/one risk factor, two risk factors, and three risk factors; particularly, the survival rate of patients with three risk factors was significantly lower than those with two risk factors and none/one risk factor. Two or more risk factors were identified in 17 of 21 (81.0%) death cases. Conclusion: The importance of serum miR‐122 and FIB‐4 index as risk factors for mortality in Japanese patients with histopathologically confirmed NAFLD is shown. Early diagnosis based on the presence of more than one risk factor and early treatment might improve the prognosis.

Immune-Related Circulating miR-125b-5p and miR-99a-5p Reveal a High Recurrence Risk Group of Pancreatic Cancer Patients after Tumor Resection.

Clinical follow-up aided by changes in the expression of circulating microRNAs (miRs) may improve prognostication of pancreatic ductal adenocarcinoma (PDAC) patients. Changes in 179 circulating miRs due to cancer progression in the transgenic KrasG12D/+; Trp53R172H/+; P48-Cre (KPC) animal model of PDAC were analyzed for serum miRs that are altered in metastatic disease. In addition, expression levels of 250 miRs were profiled before and after pancreaticoduodenectomy in the serum of two patients with resectable PDAC with different progression free survival (PFS) and analyzed for changes indicative of PDAC recurrence after resection. Three miRs that were upregulated ≥3-fold in progressive PDAC in both mice and patients were selected for validation in 26 additional PDAC patients before and after resection. We found that high serum miR-125b-5p and miR-99a-5p levels after resection are significantly associated with shorter PFS (HR 1.34 and HR 1.73 respectively). In situ hybridization for miR detection in the paired resected human PDAC tissues showed that miR-125b-5p and miR-99a-5p are highly expressed in inflammatory cells in the tumor stroma, located in clusters of CD79A expressing cells of the B-lymphocyte lineage. In conclusion, we found that circulating miR-125b-5p and miR-99a-5p are potential immune-cell related prognostic biomarkers in PDAC patients after surgery.

Serum miR-223: A Validated Biomarker for Detection of Early-Stage Non-Small Cell Lung Cancer.

The published circulating miRNA signatures proposed for early-stage non-small cell lung cancer (NSCLC) detection are inconsistent and difficult to replicate. Reproducibility and validation of an miRNA simple signature of NSCLC are prerequisites for translation to clinical application. The serum level of miR-223 and miR-29c, emerging from published studies, respectively, as a highly sensitive and a highly specific biomarker of early-stage NSCLC, was measured with droplet digital PCR (ddPCR) technique in an Italian cohort of 75 patients with stage I-II NSCLC and 111 tumor-free controls. By ROC curve analysis we evaluated the miR-223 and miR-29c performance in discerning NSCLC cases from healthy controls. Reproducibility and robust measurability of the two miRNAs using ddPCR were documented. In a training set (40 stage I-II NSCLCs and 56 controls), miR-223 and miR-29c, respectively, showed an AUC of 0.753 [95% confidence interval (CI), 0.655-0.836] and 0.632 (95% CI, 0.527-0.729) in identifying NSCLC. Combination of miR-223 with miR-29c yielded an AUC of 0.750, not improved over that of miR-223 alone. Furthermore, in an independent blind set (35 stage I-II NSCLCs and 55 controls), we validated serum miR-223 as an effective biomarker of stage I-II NSCLC (AUC = 0.808; 95% CI, 0.712-0.884), confirming the miR-223 diagnostic performance reported by others in Chinese cohorts. Using ddPCR technology, miR-223 was externally validated as a reproducible, effective serum biomarker of early-stage NSCLC in ethnically different subjects. Combination with miR-29c did not improve the miR-223 diagnostic performance. Serum miR-223 determination may be proposed as a tool for refining NSCLC risk stratification, independent of smoking habit and age.

MicroRNA 122 as adiagnostic biomarker for hepatitis C- related hepatocellular carcinoma

Aim: To explore the potential usefulness of serum microRNA(miR-122) as non-invasive diagnostic marker for hepatitis C virus(HCV)-related hepatocellular carcinoma (HCC). Methods: Thestudy has been conducted on 100 participants and were divided into 4equal groups: Group I; Twenty five normal, healthy individuals. ageand gender-matched healthy volunteers (control group). Group II;Twenty five chronic HCV patients without cirrhosis. Group III;Twenty five HCV related cirrhosis without HCC. Group IV; Twentyfive HCV-related HCC (proved radiologically by abdominal ultrasonography(US) & Triphasic abdominal computed tomography (CT). Allparticipants underwent full clinical assessment and laboratoryinvestigations in addition to the evaluation of the level of serum miR-122 by RT-PCR. Results: microRNA-122 displayed significant foldincrease in expression level in chronic hepatitis group (9.85), and lesssignificant fold increase in cirrhosis group (3.73) and significantfold increase in expression level in HCC group (7.46). Comparingserum miR-122 expression level between different studied groupsdisplayed that, No significant fold change in miR-122 expressionwas found between different groups and that miRNA122 had nosignificant up-regulation in HCC patients in comparison to non-HCCpatients (Hepatitis and Cirrhosis); (P value = 0.682). Specificity &sensitivity was 94% & 16% respectively, with AUROC (0.529).Conclusion: miR-122 could not be used as a biomarker for earlydetection of HCC in HCV related cirrhosis.

Elevated level of miR-17 along with decreased levels of TIMP-1 and IL-6 in plasma associated with the risk of in-stent restenosis.

In-stent restenosis is highly related to the deposition of inflammatory extracellular matrix and the migration of endothelial and vascular smooth muscle cells. The miR-17/TIMP-1/interleukin pathway regulates vascular matrix remodeling and plays an important role in the inflammatory reaction. This study identified miR-17 and its related biomarkers in serum that potentially indicated susceptibility to in-stent restenosis (ISR) after coronary artery stenting. Subjects were 42 patients with single de novo coronary artery lesions who underwent regular coronary angiography one year after percutaneous coronary intervention. The clinical baseline information was recorded. Serum levels of biomarkers (including miR-17, TIMP-1, IL-6, IL-8, IL-2R, TNF-alpha, IL-10, and IL-1beta) were measured with real-time PCR or ELISA. Intergroup comparisons were used to compare patients with or without ISR. Compared to levels in the non-restenosis group, the serum miR-17 level was significantly higher (3.13 ± 0.22 vs. 1.06 ± 0.04, p < 0.01) and the serum TIMP-1 and IL-6 levels were significantly lower in the ISR group (TIMP-1: 0.33 ± 0.04 vs. 1.00 ± 0.05, p < 0.01; IL-6: 1.64 ± 0.18 vs. 3.52 ± 0.11, p < 0.01). Moreover, the levels of TIMP-1 and IL-6 decreased as the level of miR-17 increased. Spearman's correlation analysis indicated that the miR-17 level was inversely correlated with TIMP-1 and IL-6 levels. Findings suggest that an elevated level of miR-17 and decreased levels of TIMP-1 and IL-6 may be associated with the risk of ISR, which is in accordance with vascular matrix remodeling and an inflammatory reaction during the pathologic process of ISR. This study highlighted the potential for miR-17, TIMP-1, and IL-6 to serve as biomarkers for ISR.

Circulating miR-130b- and miR-21-based diagnostic markers and therapeutic targets for hepatocellular carcinoma.

BackgroundHepatocellular carcinoma (HCC) is one of the histological types of primary liver cancer with high recurrence and mortality in the world. The purpose of this study was to explore the diagnostic and therapeutic value for HCC patients.MethodsIn this study, we investigated the circulating miR‐130b‐5p (miR‐130b) and miR‐21‐5p (miR‐21) expression levels in patients with HCC and their association with clinical parameters.ResultsThe circulating miR‐130b and miR‐21 were all upregulated in patients with HCC. The upregulated microRNAs (miRNAs) were associated with clinicopathological parameters of tumor capsular infiltration and clinical TNM stage. Also, the poor prognosis of patients with upregulated miRNAs levels suggested that it may be an effective therapeutic target for HCC by suppression of the miRNAs levels. In addition, the combined detection of serum miR‐130b and miR‐21 performed better in the diagnosis of HCC with a sensitivity of 92.16% and an accuracy rate of 77.51%. In vivo, tumors treated with the nanoparticle (NP)/miR‐130b and miR‐21 inhibitor complexes had significantly lower growth than the other groups.ConclusionThe circulating miR‐130b and miR‐21 can be used as potential tumor biomarkers to diagnose liver cancer, and the combined detection of serum miR‐130b and miR‐21 is superior to the diagnosis of HCC. NP/miR‐130b and miR‐21 inhibitor complexes show good therapeutic effects in vivo and are expected to become therapeutic targets worthy of further study.

Comprehensive transcriptomic analyses of tissue, serum, and serum exosomes from hepatocellular carcinoma patients

BackgroundThe expression of microRNAs (miRNAs) is a promising prognostic and diagnostic tool in hepatocellular carcinoma (HCC). Here we performed small RNA sequencing (sRNA-seq) of tissue, serum and serum exosomes to investigate changes in miRNA expression between the different sample types and correlated the expression with clinical parameters. We also performed gene expression arrays on tumor and normal tissue.ResultsPaired tissue, serum and serum exosomes sequencing revealed consistent positive correlation of miR-21 between serum exosomes and tumor tissue, indicating that miR-21 could be exported from tissue to circulation via exosomes. We found that let-7 miRNAs are generally upregulated in serum exosomes compared to whole serum, indicating that these miRNAs could be preferentially loaded into exosomes. Comparing serum from HCC patients with serum from healthy individuals revealed a global increase of miRNAs in serum from HCC patients, including an almost 4-fold increase of several miRNAs, including the liver-specific miR-122. When correlating miRNA expression with clinical parameters we detected significant association between hepatitis B virus (HBV) infection and miR-122 in serum as well as several serum and tissue-miRNAs that correlated with surgery type. We found that miR-141 and miR-146 correlated with cirrhosis in tumor tissue and normal tissue, respectively. Finally, high expression of miR-21 in tumors were associated with poor survival. Focusing on gene expression we found several significant messenger RNAs (mRNAs) between tumor and normal tissue and a Gene Ontology (GO) analysis revealed that these changes were mainly related to cell cycle and metabolism. Further, we detected mRNAs that correlated with cirrhosis and HBV infection in tissue. Finally, GO analysis of predicted targets for miRNAs down-regulated in tumor found that these were enriched for functions related to collagen synthesis.ConclusionsOur combined data point to altered miRNA and mRNA expression contributing to both generally impaired lipid metabolism and increased cell proliferation and a miRNA-driven increase in collagen synthesis in HCC. Our results further indicate a correlation in miRNA expression between exosomes, serum, and tissue samples suggesting export from tumors via exosomes. This correlation could provide a basis for a more tumor-specific miRNA profile in serum.

MiR-132 serves as a diagnostic biomarker in gestational diabetes mellitus and its regulatory effect on trophoblast cell viability

BackgroundGestational diabetes mellitus (GDM) leads to poor pregnancy outcomes. Strategies that improve trophoblast cell function are important methods for GDM treatment. This study aimed to investigate the expression and diagnostic potential of microRNA-132 (miR-132) in GDM patients, and further analyzed the effects of miR-132 on HTR-8/SVneo cell proliferation.MethodsQuantitative real-time PCR was applied to estimate the expression of miR-132. A receiver operating characteristics curve (ROC) analysis was performed to evaluate the diagnostic value of serum miR-132 in GDM patients. In vitro regulation of miR-132 in trophoblast cell HTR-8/SVneo was achieved by cell transfection, and the effects of miR-132 on cell proliferation were assessed using CCK-8 assay.ResultsExpression of miR-132 was decreased in serum and placenta tissues in GDM patients compared with the healthy women. A negative correlation was found between the serum miR-132 levels and fasting blood glucose of the GDM patients. A ROC curve shown the serum miR-132 had considerable diagnostic accuracy with an area under the curve (AUC) of 0.898. High glucose (HG) treatment induced an inhibition in HTR-8/SVneo cell proliferation and the expression of miR-132. The overexpression of miR-132 in HTR-8/SVneo cells could markedly rescued the HG - induced suppressed cell proliferation.ConclusionAll the data of this study revealed the reduced expression of miR-132 in serum and placenta tissues of GDM, and serum miR-132 serves a candidate biomarker in the diagnosis of GDM. miR-132 may act a protective role against GDM via enhancing the trophoblast cell proliferation.

Serum and tissue miRNAs: potential biomarkers for the diagnosis of cervical cancer

BackgroundFinding new biomarkers for the early detection of cervical cancer is an essential requirement in this field. In this study, we aimed to evaluate the expression level of potential biomarkers in progression of cervical cancer in patients with cervical cancer compared to normal subjects.MethodsThe expression levels of tissue and serum miRNAs, including miR-9, miR-192 and miR-205, were investigated in 36 normal, 18 precancer, and 18 cervical cancer samples using real-time polymerase chain reaction.ResultsThe results showed the higher significant expressions of miR-9, miR-192 and miR-205 in the tissue of cancer samples than those in the normal samples. Moreover, the miR-192 and miR-205 expression were significantly increased in the cancer group in comparison with the precancer group. Examination of serum samples revealed the increase in the expression level in the cancer groups than in the normal samples, for miR-9, miR-192 and miR-205 and the expressions of miR-9, miR-192 and miR-205 were significantly up-regulated in the precancer group in comparison with the normal group. Also the expression of miR-205 was remarkably increased in the cancer group in comparison with the precancer group. The receiver operating characteristic (ROC) analyses showed the highest area under the curve value for miR-192.ConclusionsGiven the increased expression level of miR-192 in cancer and in precancerous tissue and serum compared with the normal tissue and serum validated by analysing the ROC curve, miR-192 can be used as potential biomarker for the early detection of cervical cancer.

Expression and predictive value of miR-489 and miR-21 in melanoma metastasis.

BACKGROUNDMelanoma is a highly malignant skin tumour, and is one of the most rapidly growing malignant tumors in recent years. According to statistics, the morbidity of cancer increases with age, accounting for 1.6% of new cancer cases and 0.6% of deaths worldwide. Melanoma has a serious impact on society and families, thus it is of great significance to find biological markers related to the diagnosis and treatment of melanoma.AIMTo explore the expression and predictive value of mir-489 and mir-21 in melanoma metastasis.METHODSA total of 60 patients with malignant melanoma treated at our hospital from June 2017 to December 2018 were selected as a research group, while 40 healthy subjects were selected as a control group. qRT-PCR technique was used to detect miR-489 and miR-21 in serum of the two groups. ROC curve was drawn to evaluate the predictive value and diagnostic efficiency. Spearman test was used for correlation analysis. Logistic single- and multiple-factor analyses were performed to identify the risk factors related to melanoma metastasis.RESULTSThe expression of miR-489 in the research group was significantly lower than that in the control group (P < 0.001). However, the expression of miR-21 in the research group was significantly higher than that in the control group (P < 0.001). The expression of miR-489 and miR-21 was related to TNM stage and metastasis (P < 0.001). In the diagnosis of melanoma patients, the sensitivity, specificity, and AUC of miR-489 alone were 75.56%, 80.00%, and 0.852, respectively. The sensitivity, specificity, and AUC of miR-21 alone were 77.78%, 82.22%, and 0.844, respectively. MiR-489 was negatively correlated with TNM stage of melanoma (r = -0.612, P < 0.001), while miR-21 was positively correlated with TNM stage (r = 0.609, P < 0.001). Logistic single- and multiple-factor regression analyses showed that TNM stage, miR-489, and mir-21 were independent risk factors for malignant melanoma metastasis.CONCLUSIONMiR-489 and miR-21 may participate in the process of melanoma occurrence, development, and metastasis, and can be used as potential serum biomarkers for melanoma metastasis diagnosis and disease assessment.

Changes of serum inflammatory factors and miR-145 expression in patients with osteoarthritis before and after treatment and their clinical value.

BACKGROUNDOsteoarthritis is a chronic degenerative disease with an incidence of 50% in people over 65 years old and 80% in people over 80 years old worldwide. It is the second leading reason of loss of working capacity after cardiovascular diseases and severely affects the society and families. Therefore, finding biological markers related to the diagnosis and treatment of osteoarthritis is of great significance in clinical practice.AIMTo observe the changes and clinical value of serum inflammatory factors and miR-145 expression in patients with osteoarthritis before and after treatment.METHODSEighty-three patients with knee osteoarthritis (observation group) who were admitted to our hospital from April 2013 to June 2015, and 60 healthy people (control group) during the same period were selected. After 4 wk of treatment, the levels of miR-145, tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-10 were compared between the control group and the observation group before treatment. The correlation of miR-145, TNF-α, IL-6, and IL-10 levels with visual analogue scale (VAS), Lysholm, and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores was assessed by Pearson correlation analysis. The correlation of the expression of miR-145, TNF-α, IL-6, and IL-10 with Kellgren-Lawrence (K-L) grades was assessed by Spearman correlation analysis. The critical levels of miR-145, TNF-α, IL-6, and IL-10 in distinguishing different K-L grades were determined by receiver operating characteristic (ROC) curve analysis.RESULTSThe expression level of miR-145 in the observation group was significantly higher than that in the control group before treatment (P < 0.05). After treatment, the expression level of miR-145 in the observation group was significantly lower than that before treatment (P < 0.05). The levels of TNF-α and IL-6 in the observation group were significantly higher than those in the control group (P < 0.05), and the level of IL-10 was significantly lower than that in the control group (P < 0.05). After treatment, the levels of TNF-α and IL-6 in the observation group were significantly lower than those before treatment (P < 0.05), and IL-I0 level was significantly higher than that before treatment (P < 0.05). VAS and WOMAC scores were both positively correlated with miR-145, TNF-α, and IL-6 (P < 0.05), and negatively correlated with IL-10 (P < 0.05), while Lysholm scores were negatively correlated with miR-145, TNF-α, and IL-6 (P < 0.05), and positively correlated with IL-10 (P < 0.05). K-L grades were positively correlated with miR-145, TNF-α, and IL-6 (P < 0.05), and negatively correlated with IL-10 (P < 0.05). The area under the ROC curve (AUC) and specificity of TNF-α in differentiating K-L grades I-II were the highest, which were 0.785 and 97.45%, respectively, and miR145 had the highest sensitivity of 94.59%; the AUC and sensitivity of IL-6 in differentiating K-L grades II-III were the highest, which were 0.766 and 97.30%, respectively, and TNF-α had the highest specificity of 86.68%.CONCLUSIONMiR-145 and inflammatory factors have certain diagnostic value in osteoarthritis, and they are expected to become potential indicators for the diagnosis and evaluation of osteoarthritis in the future.

Elevated serum levels of brain-derived neurotrophic factor and miR-124 in acute ischemic stroke patients and the molecular mechanism.

Brain-derived neurotrophic factor (BDNF) and microRNAs (miRNAs) play a significant role in the pathogenesis of acute ischemic stroke (AIS). The present study investigates the elevated expression of BDNF and miR-124 in AIS patients. In the present study, serum samples from AIS patients and healthy controls were collected to determine the regulatory role and mechanism of operation of BDNF and to determine the regulatory miRNAs involved in AIS. Using bioinformatics analysis, we identified putative and regulatory miR-124. The effect of miR-124 on BDNF expression was examined in human neuronal cell lines. Moreover, the function of miR-124 in regulating BDNF was analyzed by assessing the serum level of BDNF in both AIS patients and healthy controls. The results indicate that the BDNF level of AIS patients is very low compared with that of controls. In contrast, real-time polymerase chain reaction (RT-PCR) data revealed a very high serum level of miR-124 in AIS patients relative to healthy individuals. The associations of the National Institutes of Health (NIH) stroke scale (NIHSS) score with BDNF and BDNF-related miR-124 serum levels were calculated using Pearson's/Spearman's correlation coefficient. The findings revealed a negative correlation between NIHSS score and BDNF level, whereas a positive correlation was observed between NIHSS score and miR-124. In addition, the relationship between serum BDNF and miR-124 was negative in AIS patients. In conclusion, this study provides strong evidence that serum BDNF and the BDNF-regulatory miR-124 may serve as molecular markers for AIS.

Poly-Phytopharm Increases Act Score and Serum miR-126 Relative Expressions, and Decreases Blood Eosinophil Levels in Asthma Patients

Background: The levels of asthma control is an indicator of asthma management outcomes. A complex immunological mechanism influences it. We aimed to investigate the effect of Poly-Phytopharm on the asthma control tests (ACT) score, blood eosinophil levels, and miR-126 relative expressions in asthma patients. &#x0D; Methods: Quasi experimental method is used in 15 stable asthma patients who were not fully controlled at the pulmonary outpatient clinic of Dr. Saiful Anwar General Hospital Malang. Assessment of ACT score, blood eosinophil levels, and miR-126 relative expressions are carried out before and after supplementation of Poly-Phytopharm that consist of Nigella sativa 100 mg, Kleinhovia hospita 100 mg, Curcuma xanthorrhiza 75 mg, and Ophiocephalus striatus 100 mg extract, three times a day, two capsules respectively for 12 weeks. The ACT was scored by the investigator by direct questioning of patients, the blood eosinophil level was measured with blood analysis, and the relative expressions of miR-126 was detected with qPCR. &#x0D; Results: There were significant increase of ACT score (18.07±2.57 to 22.06±1.83, P=0.001), decrease in blood eosinophil levels (653.15±276.15 pg/mL to 460.66±202.04 pg/mL, P=0.038), and enhancement of serum miR-126 relative expressions (1.83±1.89 to 5.89±1.34, P=0.038). &#x0D; Conclusion: The administration of Poly-Phytopharm increases ACT score, decreases blood eosinophil levels and increases serum miR-126 relative expressions in not fully controlled stable asthma patients. (J Respir Indo. 2019; 39(4): 231-7)