Expression Levels Of miR-145 Research Articles

Diagnostic and prognostic value of serum miR-145 and vascular endothelial growth factor in non-small cell lung cancer.

Previous studies have reported the diagnostic and prognostic value of serum microRNA (miR)-145 and vascular endothelial growth factor (VEGF) levels in various types of cancer; however, their clinical use in non-small cell lung cancer (NSCLC) remains unclear. The present study included 215 patients, 106 with NSCLC and 109 with other lung diseases (OLDs). miR-145 expression levels were determined using reverse transcription-quantitative PCR (RT-qPCR) and VEGF levels were measured using an ELISA. The diagnostic performance was assessed using a receiver operating characteristic curve and area under the curve (AUC) analysis. A Kaplan-Meier survival curve and Cox regression analysis were employed to evaluate the prognostic significance of the markers. The biological function of miR-145 was examined in A549 and H1792 cell lines. The effects of miR-145 on cell proliferation of NSCLC cells were evaluated by flow cytometry, and the expression levels of miR-145 and cell cycle-related genes were determined by RT-qPCR. The results revealed that miR-145 and VEGF exhibited fair diagnostic performance [AUC, 0.61 (95% CI, 0.55-0.68) and AUC, 0.64 (95% CI, 0.57-0.71), respectively]. Combining age and smoking status with miR-145 and VEGF provided the best model for differentiating patients with NSCLC from those with OLDs (AUC, 0.76; 95% CI, 0.69-0.83). Furthermore, low serum miR-145 levels were associated with poor overall survival [hazard ratio (HR), 0.48; 95% CI, 0.27-0.85], whereas high VEGF levels were not associated with poor overall survival (HR, 1.47; 95% CI, 0.81-2.68). In addition, the results of the in vitro experiments indicated that miR-145 decreased cell proliferation via the induction of cell cycle arrest. In conclusion, the findings of the present study suggested that combining miR-145 and VEGF levels with clinical risk factors may be a potential diagnostic scheme for NSCLC. In addition, serum miR-145 may be used as a prognostic marker. These results indicated that miR-145 may function as a tumor suppressor in NSCLC.

Regulation of CDK inhibitor p27 by microRNA 222 in breast cancer patients

BackgroundBreast cancer is the most common of all cancers and the second leading cause of cancer-related deaths among women worldwide. MicroRNAs regulate at least 60% of the human genes, including tumor suppressor genes and oncogenes, and can thereby affect cancer risk. In this study, the prognostic values of the CDK inhibitor p27 and miR-222 as biomarkers for breast cancer were evaluated. MethodsThe real-time quantitative polymerase chain reaction method was employed to measure the expression level of miR-222, whereas the serum levels of the CDK inhibitor p27 were measured via enzyme-linked immunosorbent assay. The levels were determined in sera from 110 participants representing three different groups. ResultsPatients with breast cancer exhibited significantly higher expression levels of miR-222 and lower levels of CDK inhibitor p27 than the control group. In addition, a statistically significant inverse correlation between miR-222 and the CDK inhibitor p27 was observed. The receiver operating characteristic curves plotted for serum p27 and miR-222 helped in significantly differentiating between breast cancer patients and controls but could not discriminate between those with stage II and stage III cancer. ConclusionThus, a significant upregulation in the serum miR-222 levels was observed in cancer patients, and a significant inverse correlation was noted between the miR-222 and CDK inhibitor p27 expression levels. These findings indicate that miR-222 may be used as a useful noninvasive screening biomarker for human breast cancer. MicroabstractNovel biomarkers for prognosis, prediction, and therapeutic purposes are essential as the prognosis and therapeutic targets of breast cancer are dependent on traditional markers, such as the tumor stage and hormonal receptor status. This study aimed to evaluate the diagnostic and prognostic values of the CDK inhibitor p27 and miR-222 in breast cancer. Our results indicated that miR-222 and the CDK inhibitor p27 may be used as noninvasive biomarkers to screen for human breast cancer but cannot discriminate between patients with early breast cancer and patients with advanced breast cancer.

Circulating miRNAs as Potential Biomarkers in Prostate Cancer Patients Undergoing Radiotherapy

IntroductionDisease recurrence is a major concern in patients with localized prostate cancer (PCa) following treatment with radiotherapy (RT), and few studies have evaluated the clinical relevance of microRNAs (miRNAs) prior and post-RT.PurposeWe aimed to investigate the significance of miRNAs in the outcomes of prostate cancer patients undergoing radiotherapy and to identify the related pathways through bioinformatics analysis.Materials and MethodsThe expression levels of miR-21, miR-106b, miR-141 and miR-375 involved in the response to radiotherapy were assessed by RT-qPCR in the serum of PCa patients (n=56) prior- and post-RT.ResultsLow expression levels of miR-106b prior-RT were associated with extracapsular extension and seminal vesicles invasion by the tumor (p=0.031 and 0.044, respectively). In the high-risk subgroup (n=47), post-RT expression levels of miR-21 were higher in patients with biochemical relapse (BR) compared to non-relapse (p=0.043). Also, in the salvage treatment subgroup (post-operative BR; n=20), post-RT expression levels of miR-21 and miR-106b were higher in patients with BR compared to non-relapse (p=0.043 and p=0.032, respectively). In the whole group of patients, high expression levels of miR-21 prior-RT and of miR-106b post-RT were associated with significantly shorter overall survival (OS; p=0.049 and p=0.050, respectively). No associations were observed among miR-141 and miR-375 expression levels with clinicopathological features or treatment outcome. Bioinformatics analysis revealed significant enrichment in DNA damage response pathways.ConclusionCirculating miRNAs prior or post-RT may hold prognostic implications in patients with PCa.

The Value of MicroRNA-375 Detection for Triaging Primary Human Papillomavirus Positive Women: A Cross-Sectional Study in a General Population.

PurposeThis study aims to validate the value of microRNA (miRNA) detection for triaging human papillomavirus (HPV)-positive women in the general population.Patients and MethodsmiR-375 detection in cervical exfoliated cells has been demonstrated to have the superior value to cytology in triaging primary HPV-positive women in the hospital population. In this study, residual samples of cervical exfoliated cells from 10,951 women in a general population were used to detect miRNA. The performance efficiency of miRNA detection in identifying high-grade cervical intraepithelial neoplasia (CIN) was evaluated. Pearson chi-square test and McNemar pairing test were used to compare miRNA detection and cytology.ResultsIn valid 9,972 women aged 25–65, miR-375 expression showed a downward trend along with an increase in cervical lesion severity. The expression level of miR-375 ≤1.0 × 10-3 was identified as positive. In the HPV-positive and 12 HPV genotypes other than 16/18 (HR12)-positive women, miR-375 detection showed equivalent sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) to that of cytology (≥ASC-US) and higher or similar sensitivity and NPV but lower specificity and PPV than that of cytology (≥ASC-H) in identifying CIN3+ and CIN2+. In HPV 16-positive women, miR-375 positivity had higher sensitivity and NPV but lower specificity and PPV than that of cytology (≥ASC-H and HSIL) in identifying CIN3+ and CIN2+. The immediate CIN3+ risk of miR-375 positivity was 19.8% (61/308) in HPV-positive, 10.8% (22/204) in HR12-positive, and 43.5% (37/85) in HPV16-positive women, respectively.ConclusionThe detection of miR-375 in cervical exfoliated cells may be an optional method for triaging primary HPV-positive women in population-based cervical cancer screening.

Circulating microRNAs and Molecular Oxidative Stress in Older Adults with Neuroprogression Disorders.

Background circulating microRNAs are potential blood biomarkers differentially expressed in many diseases including neuro depression disorders. It controls the expression of human genes and associated cellular and physiological processes in normal and diseased cells. We aimed to evaluate the potential role of circulating miRNAs and their association with both stress hormones and cellular oxidative stress in neuro depression disorders occurred among older adults. Methods a total of 70 healthy subjects were included in this study. Based upon the profile of mood states (POMS-32 score), the participants classified into two groups; healthy subjects (n =30) and depression (n =40). The expression of microRNAs; miR-124, miR-34a-5p, miR-135, and miR-451-a and their correlation with cellular oxidative stress parameters; cellular NO, genes of SOD2, CAT and iNOS, and hormones; cortisol and serotonin were estimated by a quantitative real-time RT-PCR, high-performance liquid chromatography, and ELISA Immunoassay techniques, respectively. Results depression was reported in 57.14% of the participants. The results showed a significant increase (p =0.01) in the total mood scores, and relative depression domains in older adults with depression compared to healthy controls. The relative expression levels of miR-124, miR-34a-5p significantly increased and the expression levels of miR-135, and miR-451-a significantly decreased in older adults with depression compared to healthy controls. In addition, the levels of cortisol significantly increased and serotonin (5HT) significantly reduced in all participants with depression. Cellular oxidative stress analysis for depressed subjects showed that serum NO levels and the expression of iNO gene significantly increased conversely with a decline in the molecular expression antioxidative genes; SOD2, CAT, respectively. The results showed that cellular oxidative stress parameters correlated positively with depression scores, cortisol, and negatively with cellular serotonin levels. In depressed subjects, the relative expression of microRNAs correlated positively with depression score, NO, iNOS, cortisol, and negatively associated with SOD2, CAT, and serotonin. Conclusion The combination of cellular oxidative stress and hormonal levels strongly supports a role for circulating miRNAs; miR-124, miR-34a-5p, miR-135, and miR-451-a in the regulation of depression and mood disorders among older adults. The expressed microRNAs with their related association to cellular oxidative stress and adrenal hormones are a step towards understanding the role of these small RNA molecules in the progression of depression among older adults. Thus, cellular miRNAs might have a prognostic role in the diagnosis and as a target for treatment strategies in depressed subjects.

MiR-375 Inhibits the Proliferation, Migration and Invasion of Esophageal Squamous Cell Carcinoma by Targeting XPR1.

The purpose of this study was to explore the mechanism of the miR-375/XPR1 axis in esophageal squamous cell carcinoma (ESCC) and provide a new idea for targeted therapy of ESCC. Differentially expressed genes in GEO and TCGA databases were analyzed by bioinformatics. The expression levels of miR-375 and XPR1 mRNA were detected by qRT-PCR. Protein expression of XPR1 was detected by western blot. Bioinformatics analysis and dual luciferase assay were conducted to confirm the target relationship between miR-375 and XPR1. The viability, proliferation, migration and invasion of cells in each treatment group were detected by CCK-8, colony formation, wound healing and Transwell assays. Significantly down-regulated miR-375 and remarkably up-regulated XPR1 were observed in ESCC tissue and cells. Overexpression of miR-375 inhibited proliferation, invasion and migration of ESCC cells, and greatly reduced the promoting effect of XPR1 overexpression on cell proliferation, migration and invasion. Dual luciferase assay confirmed that miR-375 targeted and inhibited XPR1 expression in ESCC. These results demonstrate the regulatory role of the miR-375/XPR1 axis in ESCC cells and provide a new potential target for the precise treatment of patients with ESCC.

MiR-124 Prevents the Microglial Proinflammatory Response by Inhibiting the Activities of TLR4 and Downstream NLRP3 in Palmitic Acid-Treated BV2 Cells.

Neuroinflammation is a mechanism by which obesity or a high-fat diet leads to cognitive impairment. MiR-124, a highly expressed microRNA in the brain, can alleviate neuroinflammation by regulating microglial activation, but its mechanism is unclear. The aim of the study was to explore whether miR-124 exerted this effect through TLR4/MyD88/NF-κB p65/NLRP3 signaling in palmitic acid-treated BV2 cells. Prepared BV2 cells were treated with palmitic acid to establish an in vitro model of a high-fat diet. An miR-124 mimic and inhibitor were adopted to upregulate and downregulate the expression of miR-124, respectively. TAK-242 and NLRP3 siRNA were used to downregulate the expression of TLR4 and NLRP3. The expression levels of miR-124, signaling proteins (TLR4, MyD88, and NF-κB p65), inflammasome markers (NLRP3 and IL-1β), and microglial activated markers (CD206, Arg-1, CD86, and iNOS) were measured by qPCR and western blotting. The pyroptosis rate was assessed using flow cytometry. First, palmitic acid upregulated TLR4/MyD88/NF-κB p65 signaling, increased NLRP3 expression, elevated the pyroptosis rate, and promoted the microglial proinflammatory response in BV2 cells. Second, the miR-124 mimic and inhibitor separately alleviated and aggravated the effect of palmitic acid on microglial activation and NLRP3 expression. The miR-124 mimic also downregulated TLR4/MyD88/NF-κB p65 signaling. Third, TAK-242 did not affect the expression of miR-124 but simulated the protective effect of the miR-124 mimic on microglial activation and NLRP3 expression. Fourth, NLRP3 siRNA also inhibited the microglial proinflammatory response in BV2 cells. MiR-124 prevented the microglial proinflammatory response through TLR4/MyD88/NF-κB p65/NLRP3 signaling in palmitic acid-treated BV2 cells.

Expression and Clinical Significance of Serum Exosomal miR-375 in Patients with Gastric Cancer

To explore exosomal miR-375 expression in gastric cancer patients and its relationship with patient prognosis. A total of 53 patients diagnosed with gastric cancer in our hospital from May 2014 to May 2016 were included as the gastric cancer group, and 46 healthy women who came to our hospital for physical examination during the same period were enrolled as the healthy group. Exosomal miR-375 expression level was detected using qRT-PCR, and the diagnostic performance and prognostic significance of exosomal miR-375 in gastric cancer were explored. The gastric cancer group showed increased exosomal miR-375 expression than the healthy group (P< 0.05); Kaplan-Meier survival analysis exhibited that serum exosomal miR-375 has an AUC of 0.778, sensitivity of 69.57%, and specificity of 75.47%, whereas Cox regression analysis showed that the miR-375 expression in exosomes was an independent risk factor affecting the prognosis of gastric cancer patients (P< 0.05). Patient with gastric cancer showed upregulated miR-375 expression in serum exosomes. Serum exosomal miR-375 was found to has positive sensitivity and specificity in the diagnosis of gastric cancer, which may be associated with poor prognosis of gastric cancer patients.

Epigenetic Silencing of SOX15 Is Controlled by miRNAs rather than Methylation in Papillary Thyroid Cancer.

Methods In this study, qRT-PCR was used to investigate the expression levels of the SOX15 gene and of miR-182, miR-183, miR-375, and miR-96 in thyroid tumors and adjacent noncancerous tissues. We also investigated the methylation status of the SOX15 promoter by methylation-specific PCR in tumors and adjacent noncancerous tissues. Results We observed a statistically significant downregulation of SOX15 expression in tumors compared to noncancerous tissue samples. The methylation levels of tumors and matched noncancerous tissues were similar, but miR-182, miR-183, and miR-375 expression levels were elevated in tumor tissues compared to noncancerous tissue samples. Conclusions Our results indicate that SOX15 gene expression is associated with the pathogenesis of papillary thyroid carcinoma (PTC), and the epigenetic control of the SOX15 gene is regulated by miRNAs rather than by promoter methylation.

HDAC6 and CXCL13 Mediate Atopic Dermatitis by Regulating Cellular Interactions and Expression Levels of miR-9 and SIRT1.

Histone deacetylase 6 (HDAC6) has been known to regulate inflammatory diseases. The role of HDAC6 in allergic skin inflammation has not been studied. We studied the role of HDAC6 in atopic dermatitis (AD) and the mechanisms associated with it. The decreased expression or chemical inhibition of HDAC6 suppressed AD by decreasing autophagic flux and cellular features of AD. AD increased expression levels of the Th1 and Th2 cytokines, but decreased expression levels of forkhead box P3 (FoxP3) and interleukin-10 (IL-10) in an HDAC6-dependent manner. CXC chemokine ligand 13 (CXCL13), which was increased in an HDAC6-depenednt manner, mediated AD. MiR-9, negatively regulated by HDAC6, suppressed AD by directly regulating the expression of sirtuin 1 (SIRT1). The downregulation or inhibition of SIRT1 suppressed AD. Experiments employing culture medium and transwell suggested that cellular interactions involving mast cells, keratinocytes, and dermal fibroblast cells could promote AD; HDAC6 and CXCL13 were found to be necessary for these cellular interactions. Mouse recombinant CXCL13 protein increased HDAC6 expression in skin mast cells and dermal fibroblast cells. CXCL13 protein was found to be present in the exosomes of DNCB-treated skin mast cells. Exosomes of DNCB-treated skin mast cells enhanced invasion potentials of keratinocytes and dermal fibroblast cells and increased expression levels of HDAC6, SIRT1 and CXCL13 in keratinocytes and dermal fibroblast cells. These results indicate that HDAC6 and CXCL13 may serve as targets for the developing anti-atopic drugs.

MicroRNA‑93 knockdown inhibits acute myeloid leukemia cell growth via inactivating thePI3K/AKT pathway by upregulating DAB2.

Acute myeloid leukemia (AML) is associated with a poor prognosis in elderly adults and currently lacks optimal treatment strategies. MicroRNAs (miRNAs or miRs) have increasingly been reported to be associated with AML progression; however, the mechanisms of action of miR-93 in AML with the involvement of disabled 2 (DAB2) are currently unknown. In the present study, miR-93 expression was assessed in patients with AML and in AML cell lines. The association between miR-93 expression and the pathological characteristics of patients with AML was analyzed. AML cells were then transfected to knockdown or overexpress miR-93 in order to elucidate its function in AML progression. The target gene of miR-93 was assessed using a dual-luciferase reporter gene assay. The expression levels of miR-93, DAB2 and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway-related proteins were measured and in vivo experiments were conducted to confirm the results. It was observed that miR-93 was highly expressed in patients with AML and in AML cells. The knockdown of miR-93 in HL-60 cells inhibited AML cell proliferation and resistance to apoptosis, while the overexpression of miR-93 in THP-1 cells led to contrasting results. Moreover, miR-93 targeted DAB2 to inactivate the PI3K/AKT pathway, and the overexpression of DAB2 reversed the effects of miR-93 on THP-1 cell growth. Tumor volume, tumor weight, and the positive expression of Ki67, survivin and p53 were increased in THP-1 cells overexpressing miR-93. On the whole, the present study demonstrates that miR-93 is highly expressed in AML cells, and that the suppression of miR-93 inhibits AML cell growth by targeting DAB2 and inhibiting the PI3K/AKT pathway.

Prognostic Implications of microRNA-155, -133a, -21 and -205 in Breast Cancer Patients' Plasma.

Breast cancer, being a heterogenous disease at the intra-tumoral and intertumoral levels, presents challenges in following the progress of the disease. Tumour-secreted aberrantly expressed miRNAs obtained from peripheral blood represent a non-invasive alternative resource for detecting and monitoring the development of the disease. This study evaluates the expression of miR-155, miR-133a, miR-21 and miR-205 as non-invasive, prognostic and follow-up markers for breast cancer. Plasma expression levels of miR-155, miR-133a, miR-21 and miR-205 were measured using real-time PCR in breast cancer patients (n=63) at presentation, healthy controls (n=25), and in post-treatment samples of 31 patients. A meta-analysis was performed using 43 studies identified from PubMed, Google Scholar and Scopus databases. Hedge's g values were used to calculate the overall effect size. Plasma miR-21 levels were higher in breast cancer patients at presentation compared to controls, while no difference was observed for miR-155, miR-133a and miR-205. These results were further supported by the meta-analysis. The altered levels of miR-155 during tamoxifen treatment indicated a potential role for miR-155 in monitoring treatment response. Further, high expressions of at least three miRNAs correlated with poor overall survival in the breast cancer patients. Plasma levels of miR-155, miR-133a, miR-21 and miR-205 may be useful as prognostic and follow-up markers for breast cancer with further validation in a large cohort of patients.

Use of cellular exosomes as a new carrier in breast cancer gene therapy.

Breast cancer is recognized as a major clinical challenge in gynecological diseases worldwide. Exosomes are small vesicles derived from multicellular bodies that are secreted by many cells into the extracellular environment and thus participate in intercellular communication through the transfer of genetic information such as encoded and non-encoded RNAs to target cells. Tumor-derived exosomes are thought to be a rich source of microRNAs (miRs) that can regulate the function of other cancer cells in the tumor microenvironment. However, the exact mechanisms by which tumor cell-derived exosomes affect their neighboring cells, as well as the bio-logical function of exosomal miRs in receptor cells, are not well understood. In this study, after the overexpression of MiR-205 in breast cancer cells (MDA-MB-231 class), cell-derived exosomes were successfully isolated and characterized by electron microscopy and dynamic light scattering. The determination of MiR-205 expression levels in exosomes secreted from engineered cells confirmed the high expression of this miR in exosomes. It was also found that the treatment of tumor exosomes carrying this miR had an apoptotic induction effect and also had a significant effect on reducing the expression of Bcl-2 gene transcript in a time-dependent manner in breast cancer cells (P < 0.001). Overall, this study suggests that exosomal transfer of tumor suppressor miRs to cancer cells could be a suitable platform for nucleic acid transfer to these cells and be highly effective in cancer treatment.

Association between miR-126, miR-21, inflammatory factors and T lymphocyte apoptosis in septic rats.

MicroRNAs (miRs) serve an important role in regulating expression levels of inflammatory factors but the underlying mechanism is still unclear. The present study aimed to observe miR-126 and miR-21 expression and apoptosis in T lymphocytes and to analyze their association with cytokine release in septic rats. The septic model rats were given intraperitoneal lipopolysaccharide (LPS) and divided into 0, 12, 24, 48 and 72 h groups. Peripheral blood was collected from each group to isolate T lymphocytes. The expression levels of miR-126 and miR-21 in T lymphocytes were observed, as well as cytokine release and apoptosis. Finally, the association between miR-126, miR-21, cytokines and apoptosis in T lymphocytes was analyzed. The release of TNF-α and IL-6 in septic rats was initially elevated but then decreased. miR-126 and miR-21 levels in T lymphocytes in septic rats were lower than those of NC rats. miR-126 and miR-21 initially decreased and then increased, whereas of apoptosis of T lymphocytes increased and then decreased, in septic rats. The expression of miR-126 was positively correlated with that of miR-21 (r=0.316; P=0.029) and negatively correlated with that of TNF-α (r=-0.480; P=0.001) and IL-6 (r=-0.626; P<0.001), as well as the apoptotic rate of T lymphocytes (r=-0.377; P=0.008). Furthermore, expression levels of miR-126 were negatively corrlated with caspase-3 expression levels (r=-0.606; P<0.001) and activity (r=-0.541; P<0.001). There was a negative correlation between miR-21 and levels of TNF-α (r=-0.311; P=0.032) and IL-6 (r=-0.439; P=0.002), as well as caspase-3 expression (r=-0.398; P=0.005) and activity (r=-0.378; P=0.008). However, there miR-126 expression was not correlated with apoptotic rate of T lymphocytes. Altered expression levels of miR-126 and miR-21 reflected the severity of inflammatory response and indicated levels of T lymphocyte apoptosis in septic rats.

Aberrant miR-10b, miR-372, and miR-375 expression in the cytobrushed samples from oral potentially malignant disorders

Background/purposeMicroRNA (miRNA) alterations play important roles in the neoplastic process of oral squamous cell carcinoma (OSCC). Upregulation of miR-10b and miR-372 and downregulation of miR-375 are frequent events in OSCC. The aberrances of these miRNAs in oral potentially malignant lesions (OPMD) were studied to determine their status during the establishment of OSCC. Materials and methodsCytobrushed sampling was used to collect epithelial cells from 11 OSCC and 34 OPMD lesions and matched normal mucosa. The expression levels of miR-10b, miR-372, and miR-375 were analyzed using quantitative reverse transcription polymerase chain reaction analysis. The clinical implications of these aberrances were further investigated. ResultsBoth miR-10b and miR-372 were upregulated in OPMD, but only miR-10b expression was upregulated in OSCC comparing to control. miR-375 was downregulated in OPMD and tended to be downregulated in OSCC. Dysplastic OPMD could be distinguished based on miR-372 expression level; miR-375 expression levels facilitated discrimination between OPMD and OSCC. The combined analysis of miR-375 and miR-372 remarkably enhanced the accuracy of differentiating OPMD from OSCC. ConclusionAberrant miR-10b. miR-372, and miR-375 expression occurs early during oral carcinogenesis. The detection of miR-372 and miR-375 expression using cytobrush samples may assist in differentiating between OPMD and OSCC.

The association of circulating miR-191 and miR-375 expression levels with markers of insulin resistance in overweight children: an exploratory analysis of the I.Family Study

BackgroundIn recent years, the exciting emergence of circulating miRNAs as stable, reproducible, and consistent among individuals has opened a promising research opportunity for the detection of non-invasive biomarkers. A firm connection has been established between circulating miRNAs and glycaemic as well as metabolic homeostasis, showing that levels of specific miRNAs vary under different physio-pathological conditions.ObjectiveIn this pilot study, we investigated the expression of candidate miRNAs, hsa-miR-191-3p and hsa-miR-375, in relation to biomarkers associated with insulin sensitivity in a subgroup (n=58) of subjects participating to the European I.Family Study, a project aimed to assess the determinants of eating behaviour in children and adolescents and related health outcomes. The sample included overweight/obese children/adolescents since overweight/obesity is a known risk factor for impaired glucose homeostasis and metabolic disorders. Biological targets of candidate miRNAs were also explored in silico.ResultsWe observed a significant association of the two miRNAs and early changes in glycaemic homeostasis, independent of covariates including country of origin, age, BMI z-score, puberty status, highest educational level of parents, total energy intake, energy from fats, energy from carbohydrates, and energy from proteins.ConclusionIdentification of circulating miRNAs associated with insulin impairment may offer novel approaches of assessing early variations in insulin sensitivity and provide evidence about the molecular mechanisms connected to early changes in glycaemic homeostasis.Trial registrationISRCTN, ISRCTN62310987. Retrospectively registered, http://isrctn.com/ISRCTN62310987

Abstract 2130: Association between miR-18a-3p, miR-143 and expression of the wild-type KRAS gene in patients with sporadic colorectal cancer

Abstract Introduction: Colorectal cancer (CRC) has the third-highest incidence and the fourth-highest mortality from cancer globally. In the genesis of CRC, mutations have been identified in the KRAS gene associated with prognosis and therapeutic management, however, the epigenetic mechanisms that modulate the expression of KRAS in CRC patients when there are no mutations in the gene are not well known. miR-18a-3p and miR-143 have been described to target KRAS and thus may regulate gene expression. Objective: To determine if there is a relationship between the expression of the wild-type KRAS gene (mRNA and protein) and the expression levels of miR-143 and miR-18a-3p in FFPE tissue samples from CRC patients. Methodology: RNA was extracted from FFPE tumor tissue samples from 51 cases of patients with CRC. Expression of wild-type KRAS mRNA, miR-143 and miR-18a-3p was evaluated by qRT-PCR and expression levels of KRAS protein by immunohistochemistry. Contingency tables, Pearson's Chi-square test and logistic regression analysis were used for data analysis. Results: Regarding the expression levels of wild-type KRAS protein, a higher number of cases with a cytoplasmic expression of the protein was identified (98%) compared to the expression in the cell membrane (31%). In 7,8% (4/51) of the cases, it was not possible to determine the mRNA expression of the KRAS gene, while in 21,3% (10/47) no expression was detected. In 78,7% of the cases in which KRAS mRNA expression was detected, low (40,4%; 19/47) and high (38,3%; 18/47) expression were categorized according to the results found. It was not evidenced by a correlation between protein expression levels and mRNA expression levels of the wild-type KRAS gene. When determining the expression levels of miR-18a-3p and miR-143, it was found that 64,7% (33/51) of cases had an absent expression of miR-18a-3p and 54,9% (28/51) of cases had an absent or very low expression for miR-143. By means of a logistic regression analysis adjusted by age, stage, sex and presence or absence of KRAS protein in the membrane, it was determined that the presence of miR-18a-3p is inversely related to KRAS mRNA levels significantly (p=0,048; OR=0,346) while miR-143 only shows a tendency to an inverse correlation with KRAS mRNA levels (p=0,119; OR=0,469). Conclusions: No correlation was found between mRNA and protein expression levels of the wild-type KRAS gene, suggesting that protein levels do not strictly depend on mRNA levels. A significant inverse association was found between miR-18a-3p and wild-type KRAS mRNA levels. Citation Format: Jehison-Alirio Herrera-Pulido, Orlando Ricaurte Guerrero, Jinneth Acosta Forero, Gustavo Hernández Suárez, Pablo Moreno-Acosta, María-Carolina Sanabria Salas, Humberto Arboleda Granados, Alfredo Romero Rojas, Martha Lucía Serrano López. Association between miR-18a-3p, miR-143 and expression of the wild-type KRAS gene in patients with sporadic colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2130.

Contribution of MIR9 on Disease Progression of Chronic Myelogenous Leukemia

We have reported that MIR9, a microRNA, is overexpressed in leukemic cells derived from a certain fraction of patients with acute myeloid leukemia (AML). The aberrant expression confers poor clinical course. These lines of our evidence suggest that overexpression of MIR9 in leukemic blasts stimulates further malignant transformation in leukemic cells. This made us analyze expression levels of MIR9 in the bone marrow cells from patients with chronic myelogenous leukemia. Among 25 patients analyzed, (chronic phase (CP): n=19, accelerated phase (AP): n=1, myeloblastic phase (BP): n=5), three in BP showed aberrantly high expression of MIR9, while none of them in CP or AP did. Furthermore, we performed targeted sequencing on the genomic DNA derived from the bone marrow cells of the same series of patients to screen for known mutations in 104 genes associated with myeloid malignancies. As a result, four patients (80%) in BC showed some numbers of gene mutations (one mutation in one patients, and two, three and six mutations in one each patient), while only four patients (21%) in CP revealed up to 2 gene mutations. Mutations in epigenetics-regulating genes such as DNMT3A were detected in patients in both CP and BP, but those in BCOR, CBL, NRAS and TP53 were only in BP. In addition, aberrant MIR9 expression was associated with multiple gene mutations in two patients in BC. These data suggested that overexpression of MIR9 could at least partly contribute to disease progression of CML. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

CBFβ promotes colorectal cancer progression through transcriptionally activating OPN, FAM129A, and UPP1 in a RUNX2-dependent manner.

Colorectal cancer (CRC) is commonly associated with aberrant transcription regulation, but characteristics of the dysregulated transcription factors in CRC pathogenesis remain to be elucidated. In the present study, core-binding factor β (CBFβ) is found to be significantly upregulated in human CRC tissues and correlates with poor survival rate of CRC patients. Mechanistically, CBFβ is found to promote CRC cell proliferation, migration, invasion, and inhibit cell apoptosis in a RUNX2-dependent way. Transcriptome studies reveal that CBFβ and RUNX2 form a transcriptional complex that activates gene expression of OPN, FAM129A, and UPP1. Furthermore, CBFβ significantly promotes CRC tumor growth and live metastasis in a mouse xenograft model and a mouse liver metastasis model. In addition, tumor-suppressive miR-143/145 are found to inhibit CBFβ expression by specifically targeting its 3'-UTR region. Consistently, an inverse correlation between miR-143/miR-145 and CBFβ expression levels is present in CRC patients. Taken together, this study uncovers a novel regulatory role of CBFβ-RUNX2 complex in the transcriptional activation of OPN, FAM129A, and UPP1 during CRC development, and may provide important insights into CRC pathogenesis.

MiR-124 Regulates the Inflammation and Apoptosis in Myocardial Infarction Rats by Targeting STAT3.

This study aimed to discover the effect of miR-124/STAT3 axis on the inflammation and cell apoptosis in myocardial infarction (MI) rats. Sprague-Dawley (SD) male rats were selected for establishing MI models and divided into Sham, MI, MI + anti-miR-124 and MI + Ad-miR-124 groups. Cardiac function was detected via echocardiography. Hematoxylin & eosin (HE) and triphenyltetrazolium chloride (TTC) staining were used to observe the pathological changes and infarction area, while transferase (TdT)-mediated D-UTP-biotin nick end labeling (TUNEL) assay was to observe myocardial apoptosis. Enzyme-linked immunosorbent assay (ELISA) was used to determine the serum levels of inflammatory cytokines. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and western blotting were performed to determine the mRNA and protein levels, respectively. Dual luciferase reporter gene assay revealed that STAT3 was a target gene of miR-124. The expression levels of miR-124 were increased and the pSTAT3/STAT3 ratio was reduced in the MI rats. The rats in the MI group showed enhanced LVEDD and LVESD, reduced LVEF and LVFS, as well as larger myocardial infarction area compared with the Sham group, Besides, IL-1β, IL-6, TNF-α and MCP-1 levels were elevated and the expressions of Bax/Bcl-2 ratio and cleaved caspase-3 were downregulated in MI group. We further found that silencing miR-124 improved cardiac function, reduced infarction area and the levels of inflammatory cytokines, as well as prevented myocardial apoptosis in MI rats. Silencing miR-124 could inhibit the inflammation and apoptosis of myocardial cells, thereby relieving the MI injury via upregulation of STAT3.