Expression Levels Of Serum miR-21 Research Articles

Pre and Postoperative Serum miR-21 Expression Levels in Oral Cancer and it’s Association with Clinical and Histological Parameters: An Observational Study

Introduction: The patterns of microRNA (miRNA) expression have opened up new avenues in the search for prognostic biomarkers and potential therapeutic targets for various tumours. In oral carcinogenesis research, analysing microRNAs expressed in Oral Squamous Cell Carcinoma (OSCC) unveils a complex network of interest. miR-21 is known to be overexpressed in numerous solid tumours and is linked to the progression of malignancies in hepatocellular carcinomas, breast cancer, and colon carcinomas. Aim: To determine the impact of surgical excision on serum miR-21 expression levels in OSCC cases and to establish correlations with clinicopathological parameters. Materials and Methods: The study was an exploratory prospective observational study conducted with proper institutional ethical approval at the Cancer Research Institute (CRI), Himalayan Institute of Medical Sciences (HIMS), Jolly Grant, Dehradun, Uttarakhand, India. Fifty-six histologically confirmed OSCC cases were enrolled along with 25 healthy subjects as controls from May 2021 to June 2023. The relative fold expression change was calculated using the Livak method (2-∆∆ct). Data analysis was performed using software such as Statistical Package for Social Sciences (SPSS) version 20.0 and MS Excel. Non parametric statistical tests such as Wilcoxon signed-rank test, MannWhitney test, and Kruskal-Wallis test were utilised. Results: Out of 56 sample included, 28 exhibited high expression of miR-21 (p-value<0.001), while 27 cases showed downregulation postsurgery (p-value<0.001), with one sample showing the same level of expression. In cases where the time difference between pre and postsurgery samples was over 25 days, no significant change was observed (p-value=0.06), and similarly, in cases with a time difference below 25 days, no significant difference was noted (p-value=0.14). A significant negative correlation was found between presurgery serum Albumin to Globulin ratio (A:G ratio) and miR-21 expression (p-value <0.05), while no other parameters showed a significant correlation with miR-21. Conclusion: miR-21 expression decreased significantly in some OSCC cases; however, in the majority of cases, it increased postsurgery. A significant correlation was observed between miR-21 expression and the serum A:G ratio.

Circulating miRNA-21 as a diagnostic biomarker in elderly patients with type 2 cardiorenal syndrome

Circulating miRNAs have attracted attention as serum biomarkers for several diseases. In this study, we aimed to evaluate the diagnostic value of circulating miRNA-21 (miR-21) as a novel biomarker for elderly patients with type 2 cardiorenal syndrome (CRS-2). A total of 157 elderly patients with chronic heart failure (CHF) were recruited for the study. According to an estimated glomerular filtration rate (eGFR) cut-off of 60 ml/min/1.73 m2, 84 patients (53.5%) and 73 patients (46.5%) were assigned to the CRS group and the CHF group, respectively. Expression levels of serum miR-21 and biomarkers for CRS, such as kidney injury factor-1 (KIM-1), neutrophil gelatinase-related apolipoprotein (NGAL), cystatin C (Cys C), amino-terminal pro-B-type natriuretic peptide (NT-proBNP), N-acetyl-κ-D-glucosaminidase (NAG), and heart-type fatty acid–binding protein (H-FABP), were detected. Serum miR-21, KIM-1, NGAL, Cys C, NT-proBNP and H-FABP levels were significantly higher in the CRS group than in the CHF group (P < 0.01), whereas NAG expression was not significantly different between the two groups (P > 0.05). Cys C, H-FABP and eGFR correlated significantly with miR-21 expression, but correlations with miR-21 were not significant for NT-proBNP, NGAL, NAG and KIM-1. Moreover, multivariate logistic regression found that serum miR-21, increased serum Cys C, serum KIM-1, hyperlipidaemia and ejection fraction (EF) were independent influencing factors for CRS (P < 0.05). The AUC of miR-21 based on the receiver operating characteristic (ROC) curve was 0.749, with a sensitivity of 55.95% and a specificity of 84.93%. Furthermore, combining miR-21 with Cys C enhanced the AUC to 0.902, with a sensitivity of 88.1% and a specificity of 83.6% (P < 0.001). Our findings suggest that circulating miR-21 has medium diagnostic value in CRS-2. The combined assessment of miR-21 and Cys C has good clinical value in elderly patients with CRS-2.

Serum miR-21 and miR-210 as promising non-invasive biomarkers for the diagnosis and prognosis of colorectal cancer.

this study aimed to investigate the expression and clinical significance of miR-21 and miR-210 in serum of patients with colorectal cancer (CRC). the expression levels of serum miR-21 and miR-210 in 40 CRC patients (CRC group) and 20 healthy patients (control group) were measured by qRT-PCR. Correlation analysis was performed of the relationship between serum miR-21 and miR-210 levels with clinical characteristics, including gender, age, tumor location, tumor size, tumor stage, local invasion and TNM staging. The expression levels of miR-21 and miR-210 in the CRC group were separately measured before and after surgery. ROC analysis was performed to evaluate the diagnostic value of miR-21 and miR-210. serum miR-21 and miR-210 in the CRC group were much higher than those in the control group. Meanwhile, the levels of serum miR-21 and miR-210 were closely related to tumor size (p = 0.028, p = 0.047), lymphatic metastasis (p = 0.038, p = 0.028), TNM staging (p = 0.014, p = 0.047) and tumor stage (p = 0.014, p = 0.017), but independent of gender, age and tumor location. In addition, serum miR-21 and miR-210 in the CRC group (n = 18) after surgery were lower than those before surgery (p < 0.001). ROC curves showed that miR-21 (AUC = 0.863) and miR-210 (AUC = 0.818) both had diagnostic efficacy in CRC patients. miR-21 and miR-210 can be used as novel non-invasive biomarkers for CRC diagnosis and prognosis.

Assessment of Serum MicroRNA-21 Gene Expression for Diagnosis and Prognosis of Colorectal Cancer.

MicroRNAs (miRNAs) are small, non-coding RNAs that are involved in carcinogenesis through posttranscriptional gene regulatory activity. The current study aimed to evaluate serum miR-21 expression levels as potential biomarkers for diagnosis and prognosis of colorectal cancer (CRC) patients. Quantitative real-time RT-PCR was applied to determine the relative expression level of miR-21 in serum. At the same time, the sensitivity and specificity of this marker were evaluated by receiver operating characteristic (ROC) curve analysis. miR-21 expression levels of serum were 3.4 and 1.25 in patient and control, respectively (p < 0.05). The sensitivity and specificity of miR-21 were found to be 95.8% and 91.7%, respectively. The high expression level of serum miR-21 were associated with higher local recurrence, TNM staging, PT staging, venous invasion, liver metastasis, and recurrence (p < 0.05). The results of this study indicated that miR-21 expression levels in serum can be considered as a novel non-invasive biomarker for early detection and prognosis of CRC patients.

MicroRNA-21 as a predictor and prognostic factor for trastuzumab therapy in human epidermal growth factor receptor 2-positive metastatic breast cancer.

Breast cancer is the second most common cancer diagnosed worldwide. Human epidermal growth factor receptor 2 (HER2)-positive breast cancer represents about 20% to 30% of all breast cancers. Trastuzumab is used in the treatment of HER2-positive breast cancer. MicroRNA-21 (miR-21) is an oncomiR that acts by inhibiting many tumor-suppressor genes. We analyzed the relative expression levels of serum miR-21 in 20 HER2-positive metastatic breast cancer patients before and after 3 months of treatment with trastuzumab. miR-21 levels decreased with a high significant difference after trastuzumab therapy (P = 0.001). Although miR-21 expression levels were lower in responders than in nonresponders, the difference was not statistically significant ( P = 0.6). Our results demonstrated a significant negative correlation between its basal expression, expression levels after treatment, and time to progression ( P = 0.03 and 0.01, respectively). These results make miR-21 a potential prognostic factor for HER2-positive metastatic breast cancer patients. Additionally, it can be an interesting potential target in therapy using antisense oligonucleotides for miR-21.

Clinical evaluation of 4 types of microRNA in serum as biomarkers of esophageal squamous cell carcinoma.

To the best of our knowledge, there is currently no specific biomarker for esophageal cancer used in clinical practice. However, studies consider that microRNAs (miRNAs/miRs) could have useful implications in clinical practice. The present study aimed to investigate the feasibility of using serum microRNAs as biomarkers for esophageal squamous cell carcinoma (ESCC). Using reverse transcription-quantitative polymerase chain reaction, the expression levels of serum miR-21, miR-25, miR-145 and miR-203 were detected in 31 untreated patients with ESCC (EC-UT), 35 inactive period patients with ESCC following treatment (EC-T), 33 patients with esophageal benign disease (benign) and 32 healthy donors (healthy). Furthermore, the ability of these microRNAs to function as biomarkers of ESCC alone and in combination were investigated. The expression levels of serum miR-21, miR-25 and miR-145 in EC-UT were significantly higher than in the other groups (P<0.001). High sensitivity and specificity were shown when miRNAs were used as biomarkers for ESCC, particularly miR-21 and the combination of miR-21 with miR-145. Comparing EC-UT with healthy, benign and EC-T groups, and a combined group (3 groups set as 1 negative control), the sensitivity and specificity of miR-21 were 71.0 and 96.9, 74.2 and 87.9, 77.4 and 82.9, and 74.2 and 88.0%, respectively. The combined sensitivity and specificity of miR-21 and miR-145 were 71.0 and 96.9, 90.9 and 72.7, 97.1 and 82.9, and 80.6 and 80.0%, respectively. In conclusion, 3 types of miRNA (miR-21, miR-25 and miR-145) in serum could serve as potential biomarkers for ESCC. Furthermore, the expression level of miR-145 in serum was upregulated, compared with the downregulation reported in previous studies in ESCC tissues and cells.

Circulating microRNA-21 as a prognostic, biological marker in cholangiocarcinoma.

The prognosis of cholangiocarcinoma (CCA) is generally poor because there is a lack of effective diagnostic tools including laboratory assessments and imageological examination. Therefore, a novel biological marker (biomarker) to effectively diagnose cancer is highly desirable in clinical. Previously, serum microRNAs as biomarkers of cancers have been reported. However, it was still unclear about the clinical significance of serum microRNA-21 (miR-21) expression levels for CCA. The serum samples were separately collected from fifty patients of CCA, 15 patients of hepatolithiasis, and 15 healthy volunteers; quantitative real-time polymerase chain reaction was used for measuring miR-21 expression level in serum. The clinicopathological data were recorded before patients discharged. In the CCA serum, the expression level of miR-21 significantly upregulated (P < 0.05). With the tumor, node, and metastasis stage developed (Stage I vs. III and IV, P < 0.05), the serum miR-21 expression level increased, but there was no statistical difference between Stage I patients and hepatolithiasis patients or healthy control (P > 0.05 for both). Furthermore, the miR-21 level was significant difference between pre- and post-operative serum (P < 0.05) for the high miR-21 expression group. The serum miR-21 expression levels were defective in discriminating patients with CCA from healthy control subjects by receiver-operator curve analysis because the area under the curve (AUC) value was 0.871 which was not better than the conventional CCA markers-carbohydrate antigen 19-9 (AUC value = 0.96). However, in serum, high expression level miR-21 was significantly related to clinical stage, invasion depth, lymph vessel infiltration, metastasis status, differentiation status, whether to resection, and poor survival of CCA patients (P < 0.05 for all). This study suggested that serum miR-21 was a promising biomarker for diagnosing the late stage CCA and would have potential to be a useful prognostic biomarker of CCA.

Diagnostic value of circulating miR-21: An update meta-analysis in various cancers and validation in endometrial cancer.

MiR-21 has been identified as one of the most common proto-oncogenes. It is hypothesized that up-regulated miR-21 could be served as a potential biomarker for human cancer diagnosis. However, inconsistencies or discrepancies about diagnostic accuracy of circulating miR-21 still remain. In this sense, miR-21′s diagnostic value needs to be fully validated. In this study, we performed an update meta-analysis to estimate the diagnostic value of circulating miR-21 in various human cancers. Additionally, we conducted a validation test on 50 endometrial cancer patients, 50 benign lesion patients and 50 healthy controls. A systematical literature search for relevant articles was performed in Pubmed, Embase and Cochrane Library. A total of 48 studies from 39 articles, involving 3,568 cancer patients and 2,248 controls, were included in this meta-analysis. The overall sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR) and area under the curve (AUC) were 0.76 (0.71-0.80), 0.82 (0.79-0.85), 4.3 (3.6-5.1), 0.29 (0.24-0.35), 15 (11-20) and 0.86 (0.83-0.89), respectively. In the validation test, the expression levels of serum miR-21 were significantly higher in benign lesion patients (p = 0.003) and endometrial cancer patients (p = 0.000) compared with that of healthy controls. Endometrial cancer patients showed higher miR-21 expression levels (p = 0.000) compared with benign lesion patients. In conclusion, the meta-analysis shows that circulating miR-21 has excellent performance on the diagnosis for various cancers and the validation test demonstrates that serum miR-21 could be served as a novel biomarker for endometrial carcinoma.

Quantitative measurement of serum microRNA-21 expression in relation to breast cancer metastasis in Chinese females.

BackgroundBreast cancer is the most common type of cancer in females. Aberrant expression of microRNA-21 (miR-21) has previously been reported in breast cancer tissue. The aim of this study was to investigate expression levels of serum miR-21 in breast cancer patients and evaluate its prognostic value in Chinese females.MethodsReal-time quantitative (RQ)-PCR was used to analyze miR-21 expression in archived serum, tumor tissue, and adjacent normal tissue from 549 participants (326 with breast cancer, 223 without breast cancer). We also analyzed associations between serum miR-21 expression and breast cancer subtypes and patient prognosis. Recurrence and survival were analyzed by using the multivariate Cox proportional hazards model.ResultsExpression of miR-21 was significantly higher in breast cancer tissues compared with normal adjacent breast tissues (P<0.001). The 2-ΔΔCt values for serum miR-21 in breast cancer patients versus healthy controls were 9.12±3.43 and 2.96±0.73, respectively. Multivariate Cox proportional hazards model suggested that serum miR-21 expression was an independent poor prognostic factor for both recurrence (hazard ratio [HR]= 2.942; 95% confidence interval [CI]=1.420-8.325; P=0.008) and disease-free survival (HR=2.732; 95% CI=1.038-7.273, P=0.003) in breast cancer.ConclusionsIncreased serum miR-21 expression level was correlated with poor prognosis of breast cancer patients, indicating that serum miR-21 may be a novel prognostic marker for recurrence and survival of breast cancer patients before resection.

Serum expression levels of miR-17, miR-21, and miR-92 as potential biomarkers for recurrence after adjuvant chemotherapy in colon cancer patients.

The present study examined whether miR-17, miR-21, miR-29a, and miR-92 that are dysregulated in colon cancer (CC) can serve as potential predictive markers for relapse of disease after radical surgery and adjuvant chemotherapy. Real-time reverse transcription quantitative polymerase chain reaction was used to measure the expression levels of the miRNAs in serum samples from 37 patients with CC and 7 healthy individuals, tested as a control group. The area under the receiver operating characteristic curve (AUC) was then used to evaluate the predictive performance of the four miRNAs alone or in combination and compare it with carcinoembryonic antigen. The expression of miR-17, miR-21 and miR-92 were significantly higher in serum of patients with disease relapse. The AUCs for miR-17, miR-21, miR-92 for Nx patients were 0.844, 0.948, and 0.935, respectively (p < 0.05). Combining the four miRNAs for stage III patients increased the diagnostic performance, yielding an AUC of 0.881, with a sensitivity of 83.3% and a specificity of 85.7% (p < 0.05). Our study suggests that the expression levels of serum miR-21, miR-17, and miR-92 in patients with CC who underwent radical surgery and adjuvant chemotherapy may have diagnostic value for differentiating between recurred and non-recurred patients.

Prognostic implications of serum microRNA-21 in colorectal cancer

MicroRNAs (miRNAs) are small, noncoding RNAs that are involved in carcinogenesis through postranscriptional gene regulatory activity. Few studies have focused on the detection of miR-21 in serum rather than in tissue. The current study aimed to measure serum miR-21 expression levels and to evaluate their association with the outcome of colorectal cancer (CRC). Blood samples were collected from 102 CRC patients undergoing surgery with curative intent. The expression levels of miR-21 were measured using a quantitative reverse transcription polymerase chain reaction (qRT-PCR). The results were analysed to assess the relationship between serum miR-21 levels and patient survival. A univariate analysis revealed that lower expression levels of serum miR-21 were associated with higher local recurrence (P = 0.025) and mortality (P = 0.029). A logistic regression analysis demonstrated that the relative overexpression of miR-21 (expression > 1) was associated with a 51% reduction in the risk of recurrence. A Cox regression analysis identified miR-21 expression as an independent predictor of survival (P = 0.048); a relative increase in miR-21 expression (>1) was associated with a 50% reduction in the risk of mortality. The expression level of serum miR-21 correlates with the recurrence and mortality of CRC patients. Our results suggest that circulating serum miR-21 is a promising prognostic tumour marker, and they highlight the potential clinical utility of miR-21 expression as a prognostic marker for CRC prognosis.

Differential microRNA expression in renal cell carcinoma

The present study aimed to detect microRNA expression levels in the tissues and sera of patients with clear cell renal cell carcinoma (ccRCC). The association of microRNA expression with ccRCC clinical pathology was analyzed, and the potential of the microRNAs as ccRCC serum markers and the significance of their expression in the clinical diagnosis, staging, prognosis and selection of new therapeutic targets for ccRCC were discussed. Specific microRNAs were selected according to the associated literature. TaqMan quantitative polymerase chain reaction (qPCR) technology was used to determine the expression levels of selected microRNAs. miR-34a, miR-224 and miR-21 were upregulated, whereas miR-141, miR-149 and miR-429 were downregulated in the ccRCC tissues (P<0.01). The expression of miR-221 and miR-211 was not significant in the ccRCC tissues (P>0.05). miR-34a, miR-21 and miR-224 were upregulated and miR-141 was downregulated in the sera of patients with ccRCC (P<0.01), while the expression of miR-149 and miR-429 was not significant (P>0.05). The serum miR-21 expression levels were significantly correlated with the clinical staging of the patients with ccRCC (P<0.05). miR-34a, miR-21 and miR-224 are upregulated in the tissues and sera of patients with ccRCC, whereas miR-141 is downregulated. miR-21 and miR-141 are associated with ccRCC and are, thus, potential ccRCC serum markers.

Abstract 4145: Serum miR-21 is a promising biomarker for early detection and prognosis of colorectal cancer

Abstract Background: MicroRNAs (miRNAs) are rapidly emerging as potential diagnostic and prognostic markers of human cancer. Recent studies indicate that tumor-derived miRNAs can also be present in human serum and plasma in remarkably stable form. Therefore, specific miRNAs that are frequently over-expressed in cancer tissues present with logical targets for the development of noninvasive miRNA biomarkers in blood. The aim of this study was to investigate whether serum miR-21 can be used as a biomarker for the earlier diagnosis and prognosis of colorectal cancer (CRC). Materials &amp; Methods This study was divided into two steps. The first step included determination of miR-21 expression using quantitative TaqMan assays in a ‘training set’ comprised of pre-operative serum samples from 12 CRC patients and 12 healthy subjects. To enhance assay specificity, we analyzed miR-21 expression in matched CRC and normal tissues from 8 of the 12 patients. The second step included an independent confirmation of these results in a ‘validation set’ of serum samples from 184 CRC patients, 43 patients with advanced adenomas and 53 healthy controls. We then examined miR-21 expression in matched normal and cancer tissue samples from 167 CRC patients. In addition, we analyzed post-operative serum samples from 60 CRC patients. Results: In the training set, miR-21 expression was significantly up-regulated in both serum (p&amp;lt;0.001) and tissue samples (p&amp;lt;0.01) of patients with CRC compared to healthy controls. In the validation set, serum miR-21 expression levels were significantly elevated in advanced adenoma (p&amp;lt;0.001) and CRC (p&amp;lt;0.001) patients. ROC analysis revealed high sensitivity and specificity for serum miR-21 in discriminating advanced adenomas (AUC: 0.8, sensitivity: 76%, specificity: 82%) and CRC (AUC: 0.92, sensitivity: 90%, specificity: 83%) from control subjects. In addition, miR-21 levels in serum closely correlated with those in CRC tissues (p&amp;lt;0.001). Of interest, surgical treatment in CRC patients resulted in significantly lowered miR-21 expression levels in the post-operative serum samples (p&amp;lt;0.001). Higher serum miR-21 levels were associated with larger tumor size (p=0.004), distant metastases (p=0.01) and higher TNM stage (p=0.04). High expression of miR-21 in both serum and primary CRC tissues were significantly associated with poor survival (serum, p=0.005; tissue, p=0.006). Serum miR-21 was an independent prognostic marker in CRC patients (HR=4.1, p=0.03). Conclusions: Since serum miR-21 expression levels accurately discriminated between healthy subjects and patients with advanced adenomas and early CRC, it may serve as a promising diagnostic biomarker for the early detection of colorectal neoplasia. Because serum miR-21 levels were a better predictor for poor survival, detection of this non-coding miRNA in serum also has potential for development as a noninvasive prognostic biomarker in CRC. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4145. doi:1538-7445.AM2012-4145

Identification of Suitable Reference Genes for qPCR Analysis of Serum microRNA in Gastric Cancer Patients

Circulating microRNA expression profiles may be promising biomarkers for diagnosis and assessment of the prognosis of cancer patients. Quantitative polymerase chain reaction (qPCR) is a sensitive technique for estimating expression levels of circulating microRNAs. However, there is no current consensus on the reference genes for qPCR analysis of circulating microRNAs. In this study we tried to identify suitable reference genes for qPCR analysis of serum microRNA in gastric cancer patients and healthy individuals. Six microRNAs (let-7a, miR-16, miR-93, miR-103, miR-192, and miR-451) and RNU6B were chosen as candidate reference genes on the basis of the literature. Expression levels of these candidates were analyzed by qPCR in serum samples from 40 gastric cancer patients and 20 healthy volunteers. The geNorm, Normfinder, bestkeeper, and comparative delta-Ct method algorithms were used to select the most suitable reference gene from the seven candidates. This was then validated by normalizing the expression levels of serum miR-21 across all gastric cancer patients and healthy volunteers. The algorithms revealed miR-16 and miR-93 were the most stably expressed reference genes, with stability values of 1.778 and 2.213, respectively, for serum microRNA analysis across all the patients and healthy controls. The effect of different normalization strategies was compared; when normalized to the serum volume there were no significant differences between patients and controls. However, when the data were normalized to miR-93, miR-16, or miR-93 and miR-16 combined, significant differences were detected. Our results demonstrated that reference gene choice for qPCR data analysis has a great effect on the study outcome, and that it is necessary to choose a suitable reference for reliable expression data. We recommend miR-16 and miR-93 as suitable reference genes for serum miRNA analysis for gastric cancer patients and healthy controls.