Underexpression of miR-126-3p in Patients with Cholangiocarcinoma

Objectives: To evaluate the expression of miR-126-3p and its potential as a biomarker for cholangiocarcinoma (CCA) and to better understand the prognosis, comorbidities, and lifestyle habits associated with the disease. Methods: Fifty-nine individuals were distributed into either the study group (38 CCA patients) or the control group (21 individuals without liver diseases). Total RNA was extracted, cDNA synthesis was performed, and miR-126-3p expression was assessed using real-time PCR. For statistical analysis, alpha error was set at 5%. Results: MiR-126-3p was found to be underexpressed in the study group relative to the controls (0.42; P=0.001). Additionally, marked underexpression was found in the study group in when associated with smoking (0.28; P=0.0001), alcoholism (0.19; P=0.0001), hypertension (0.29; P=000.1), and diabetes (0.12; P=0.0003) relative to the controls. No association was found between miR-126-3p expression and tumor subtypes (iCCA=0.42; pCCA=0.45; dCCA=0.72; P=0.9155). A total of 67% of dCCA patients were event-free at 16 months of follow up, while both pCCA and iCCA exhibited event-free survival rates of 25%, though there was no significant difference between these subgroups (P=0.273). Conclusion: The underexpression of mir-126-3p is associated with cholangiocarcinoma and can be potentiated by alcoholism, hypertension, diabetes, and smoking, the latter of which is an independent risk factor for this cancer. Furthermore, dCCA patients exhibit higher survival rates relative to patients with pCCA and iCCA.     

血清微小RNA(miR-129-3p、miR-767-3p和miR-877)在结直肠癌诊断中的价值

目的:探讨血清微小RNA (microRNA,mi RNA)在结直肠癌诊断中的价值.方法:通过miRNA表达谱芯片检测7例结直肠癌患者血清和10例健康志愿者血清中差异表达的miRNA.应用实时荧光定量PCR法在40例结直肠癌患者血清和18例健康志愿者血清中验证芯片结果,并分析血清特异性miRNA在结直肠癌诊断中的价值.结果:筛选获得10个在结直肠癌中特异性表达的血清miRNA,实时荧光定量PCR验证后获得一组结直肠癌特异性血清miRNA(miR-129-3p、miR-767-3p及miR-877*)生物标志物,这组生物标志物组合检测结直肠癌的灵敏度为77.78％、特异度为100％,并可产生最大受试者工作特征曲线(receiver operator characteristic curve,ROC)的曲线下面积(area under the curve,AUC)为0.914.结论:miR-129-3p、miR-767-3p和miR-877*生物标志物组合有望成为结直肠癌筛查和早期诊断的指标.     

Circulatory miR-628-5p is downregulated in prostate cancer patients

Prostate cancer (PCa) is a major health concern for men in the USA. Aberrant expression of microRNAs (miRNAs) has been associated with the pathogenesis of various cancers, including PCa. Circulatory forms of miRNAs have been detected in serum and hold promise as minimally invasive cancer biomarkers. This study aimed to identify potential circulatory miRNAs that can provide insights into new mechanisms for clinical diagnosis of PCa and can serve as potential biomarkers and/or therapeutic targets. Candidate serum miRNAs were detected by using PCR microarray in a learning set of six African American (AA) and six Caucasian American (CA) PCa patients. Discriminating performance of candidate miRNAs was validated by qRT-PCR in serum samples from 36 AA (24 PCa patients and 12 controls) and 36 CA (16 PCa patients and 20 controls). From the miRNA profiling experiments, three differentially expressed miRNAs (miR-25, miR-101, and miR-628-5p) were selected for future validation. In the validation set, there was an overall low expression of miR-25 (p?<?0.01), miR-101 (p?<?0.001), and miR-628-5p (p?<?0.0001) in serum of PCa patients as compared with normal individuals. Subdivision on the basis of ethnicity showed that serum expression levels of miR-628-5p were significantly downregulated in both AA and CA PCa patients when compared with their respective controls. Our results demonstrate that the three miRNAs, particularly miR-628-5p, may be further developed as a biomarker, which can serve as novel noninvasive biomarker for PCa diagnosis and prognosis.     

鼻咽癌患者血清中miR-141-3p和miR-155-3p的表达水平及其临床意义

目的:探讨鼻咽癌患者血清中miR-141-3p及miR-155-3p的表达水平及临床意义.方法:选取2017年01月至2020年09月我院收治的95例鼻咽癌患者和45例健康对照组作为研究对象,采用实时荧光定量PCR法检测miR-141-3p及miR-155-3p表达水平.应用受试者工作特征(receiver opera...     

结肠癌细胞中miR-145-5p、miR-143-3p的表达变化对多药耐药的影响

目的:探讨人结肠癌细胞中miR-145-5p、miR-143-3p的表达发生变化后对P-gp和MRP的影响。方法:采用Western Blot和qRT-PCR法检测人结肠癌细胞分别转染miR-145-5p mimics/inhibitor、miR-143-3p mimics/inhibitor后P-gp 和MRP的表达。结果:转染miR-145-5p mimics后P-gp和MRP的蛋白和基因表达均降低（P<0.05）,转染miR-145-5p inhibitor后P-gp 和MRP的蛋白和基因表达均升高（P<0.05）。转染miR-145-5p mimics NC和miR-145-5p inhibitor NC后P-gp 和MRP的蛋白和基因表达无明显变化（P>0.05）。转染miR-143-3p mimics后P-gp和MRP的蛋白和基因表达均降低（P<0.05）,miR-143-3p inhibitor后P-gp和MRP的蛋白和基因表达均升高（P<0.05）。转染miR-143-3p mimics NC和miR-143-3p inhibitor NC后P-gp 和MRP的蛋白和基因表达无明显变化（P>0.05）。结论:miR-145-5p及miR-143-3p对多药耐药蛋白P-gp、MRP具有调控作用,参与了结肠癌的多药耐药。     

MiR-19b-3p and miR-101-3p as potential biomarkers for prostate cancer diagnosis and prognosis

Prostate cancer (PCa) is the most commonly diagnosed male malignancy worldwide. Early diagnosis and metastases detection are crucial features to diminish patient mortality. High fat diet (HFD) and metabolic syndrome increase PCa risk and aggressiveness. Our goal was to identify miRNAs-based biomarkers for PCa diagnosis and prognosis associated with HFD. Mice chronically fed with a HFD or control diet (CD) were subcutaneously inoculated with androgen insensitive PC3 cells. Xenografts from HFD-fed mice showed increased expression of 7 miRNAs that we named “candidates” compared to CD-fed mice. These miRNAs modulate specific metabolic and cancer related pathways. Using bioinformatic tools and human datasets we found that hsa-miR-19b-3p and miR-101-3p showed more than 1,100 validated targets involved in proteoglycans in cancer and fatty acid biosynthesis. These miRNAs were significantly increased in the bloodstream of PCa patients compared to non-PCa volunteers, and in prostate tumors compared to normal adjacent tissues (NAT). Interestingly, both miRNAs were also increased in tumors of metastatic patients compared to tumors of non-metastatic patients. Further receiver-operating characteristic (ROC) analysis determined that hsa-miR-19b-3p and hsa-miR-101-3p in serum showed poor predictive power to discriminate PCa from non-PCa patients. Hsa-miR-19b-3p showed the best score to discriminate between tumor and NAT, while hsa-miR-101-3p was useful to differentiate between metastatic and non-metastatic PCa patients. Hsa-miR-101-3p was increased in exosomes isolated from blood of PCa patients. Although more detailed functional exploration and validation of the molecular mechanisms are required, we identified hsa-miR-19b-3p and hsa-miR-101-3p with high potential for PCa diagnosis and prognosis.     

长链非编码RNA XIST和miR-101-3p在子宫颈癌组织中的表达及关系

目的：探讨长链非编码RNA（LncRNA）XIST和miR-101-3p在子宫颈癌组织中的表达情况及其临床意义。方法：收集58例子宫颈癌和配对的癌旁组织标本,采用实时荧光定量PCR（qPCR）检测组织中LncRNA XIST和miR-101-3p的表达,分析其与子宫颈癌临床病理因素的相关性,并评价其临床意义。结果：与癌旁组织比较,qPCR显示宫颈癌组织中LncRNA XIST的表达明显上调（P < 0.05）,miR-101-3p的表达明显下调（P < 0.05）;且二者在表达水平上呈负相关（r=-0.68,P=0.000）。LncRNA XIST和miR-101-3p的表达均与宫颈癌淋巴结转移、组织大小及临床分期之间存在相关性（P均<0.05）。结论：LncRNA XIST与miR-101-3p的表达呈负相关,可能成为早期筛选宫颈癌的生物标志物和治疗靶点。     

MiR-16-1-3p and miR-16-2-3p possess strong tumor suppressive and antimetastatic properties in osteosarcoma

Osteosarcoma (OS) is an aggressive malignancy affecting mostly children and adolescents. MicroRNAs (miRNAs) play important roles in OS development and progression. Here we found that miR-16-1-3p and miR-16-2-3p “passenger” strands, as well as the “lead” miR-16-5p strand, are frequently downregulated and possess strong tumor suppressive functions in human OS. Furthermore, we report different although strongly overlapping functions for miR-16-1-3p and miR-16-2-3p in OS cells. Ectopic expression of these miRNAs affected primary tumor growth, metastasis seeding and chemoresistance and invasiveness of human OS cells. Loss-of-function experiments verified tumor suppressive functions of these miRNAs at endogenous levels of expression. Using RNA immunoprecipitation (RIP) assays, we identify direct targets of miR-16-1-3p and miR-16-2-3p in OS cells. Moreover, validation experiments identified FGFR2 as a direct target for miR-16-1-3p and miR-16-2-3p. Overall, our findings underscore the importance of passenger strand miRNAs, at least some, in osteosarcomagenesis.     

miR-455-3p在胃癌中的表达及其与MVD和VEGF表达的相关性研究

目的 探讨miR-455-3p在胃癌中的水平及其与微血管密度（MVD）和血管内皮生长因子（VEGF）表达的关系。方法 采用实时荧光定量PCR检测80例胃癌组织（胃癌组）中miR-455-3p水平,免疫组化法检测80例胃癌组织中MVD和VEGF表达,并分析胃癌组织中miR-455-3p水平与MVD和VEGF表达的相关性。选取同期的37例浅表性胃炎和12例正常胃黏膜组织作对照（对照组）。结果 胃癌组的miR-455-3p水平为1.16±0.59,低于对照组的2.61±0.88,差异有统计学意义（P＜0.05）;胃癌组的VEGF阳性表达率和MVD分别为71.3％（57／80）和54.9±7.3,均高于对照组的28.6％（14／49）和27.5±6.1,差异均有统计学意义（P＜0.05）;胃癌组织中miR-455-3p水平与VEGF表达（r=-0.783,P＜0.05）和MVD（r=-0.824,P＜0.05）均呈负相关。结论 miR-455-3p在胃癌中为低表达,并与VEGF表达和MVD呈负相关,有可能在调控胃癌组织血管生长中发挥重要作用。     

miR-331-3p regulates expression of neuropilin-2 in glioblastoma

Aberrant expression of microRNAs (miRNAs), a class of small non-coding regulatory RNAs, has been implicated in the development and progression of high-grade gliomas. However, the precise mechanistic role of many miRNAs in this disease remains unclear. Here, we investigate the functional role of miR-331-3p in glioblastoma multiforme (GBM). We found that miR-331-3p expression in GBM cell lines is significantly lower than in normal brain, and that transient overexpression of miR-331-3p inhibits GBM cell line proliferation and clonogenic growth, suggesting a possible tumor suppressor role for miR-331-3p in this system. Bioinformatics analysis identified neuropilin-2 (NRP-2) as a putative target of miR-331-3p. Using transfection studies, we validated NRP-2 mRNA as a target of miR-331-3p in GBM cell lines, and show that NRP-2 expression is regulated by miR-331-3p. RNA interference (RNAi) to inhibit NRP-2 expression in vitro decreased the growth and clonogenic growth of GBM cell lines, providing further support for an oncogenic role for NRP-2 in high-grade gliomas. We also show that miR-331-3p inhibits GBM cell migration, an effect due in part to reduced NRP-2 expression. Finally, we identified a significant inverse correlation between miR-331-3p and NRP-2 expression in The Cancer Genome Atlas GBM cohort of 491 patients. Together, our results suggest that a loss of miR-331-3p expression contributes to GBM development and progression, at least in part via upregulating NRP-2 expression and increasing cell proliferation and clonogenic growth.     

Clinical Potentials of miR-576-3p,miR-613, NDRG2 and YKL40 in Colorectal Cancer Patients

Introduction: Colorectal cancer (CRC) is the most common type of gastrointestinal tract cancers. This investigation aim was to assess the expression of miR-576-3p and miR-613 in CRC patients in addition to NDRG2 and YKL40 serum levels determination to decide their diagnostic and prognostic significance. Methods: Sixty early diagnosed CRC patients prior to any treatment in addition to twelve healthy subjects were enrolled in this study. Blood samples were taken from subjects and allowed for clotting and centrifugation, then the collected sera were stored at -80ºC till it were used for detection of our molecular biomarkers. The mature miRNAs expressions (miR-576-3p and miR-613) were detected in serum by qRT-PCR, while NDRG2 and YKL40 serum levels were determined by ELISA. In addition, the correlation of the measured parameters with the clinicopathological data of the patients was investigated. Results: The study results showed that both miRNA-576-3p and miRNA-613 were down-regulated in CRC patients with fold change 0.33, 0.36; respectively. A significant positive correlation was observed between miR-576-3p and miR-613 (r = 0.75, p < 0.001). NDRG2 serum levels were decreased in patients compared to the control group but the decrease wasn’t statistically significant. On the other hand, it was observed that YKL40 serum level was significantly increased in CRC patients compared to control (p-value < 0.001). Furthermore, YKL40 showed a very high diagnostic value (AUC = 0.97, specificity = 91.7%, sensitivity = 96%, p-value = 0.0001). Conclusion: The observations of this investigation concluded that, the expressions of miR-576-3p and miR-613 in addition to YKL40 serum levels determinations may help in the diagnosis of CRC.     

miR-590-5p, miR-219-5p, miR-15b and miR-628-5p are commonly regulated by IL-3, GM-CSF and G-CSF in acute myeloid leukemia

Aberrations in IL-3, GM-CSF and G-CSF induced signaling are frequently reported in acute myeloid leukemia (AML). Herein, we utilized a unique human myeloid leukemic cell line, AML-193, which responds to all three cytokines to analyze the regulation at microRNA level. Using real-time PCR-based miRNA expression profiling, we investigated miRNA signatures regulated by IL-3, GM-CSF and G-CSF for n=704 miRNAs. We discovered that in addition to regulating specific miRNAs, these cytokines also regulate common set of miRNAs, which includes miR-590-5p, miR-219-5p, miR-15b and miR-628-5p. Taken together, we have identified novel candidate miRNAs that may be instructive during leukemic and normal hematopoiesis.     

miR-363-3p和miR-5100在非小细胞肺癌中的表达及其临床意义

目的:探讨小分子RNA(microRNA)miR-363-3p 和miR-5100在非小细胞肺癌中的表达及其临床意义.方法:利用荧光定量PCR的方法,检测肿瘤组织、癌旁组织及远离肿瘤的正常组织中致癌基因Myc的mRNA 和miR-363-3p、miR-5100的表达,然后分析miR-363-3p 和miR-5100与临床病理特征之间的相关性.结果:Myc在肿瘤组织中表达明显升高;miR-363-3p在肺癌组织中的表达明显低于正常组织(P<0.001),而在癌旁组织中的表达却远远高于正常组织(P<0.05);miR-5100在肺癌组织中的表达显著地高于正常组织(P<0.001),并且癌旁组织中的表达也高于正常组织(P<0.05).临床相关性分析显示,miR-363-3p的表达与淋巴结转移呈正相关(P<0.001),miR-5100的表达与临床分期也呈正相关性(P<0.05).结论:miR-363-3p 和miR-5100可能参与了非小细胞肺癌早期的发生和转移,并可能作为早期诊断和预后的分子标志物.     

miR-101-3p在胃癌中的表达及其靶向STC-1基因调控PI3K/AKT信号通路对癌细胞侵袭转移和血管生成的影响

目的 探究miR-101-3p在胃癌中的表达及其靶向STC-1基因调控PI3K/AKT信号通路对癌细胞侵袭转移、血管生成的作用机制。方法 qRT-PCR检测胃癌组织及BGC-823细胞中miR-101-3p和STC-1 mRNA的表达,并分析miR-101-3p表达与患者临床病理因素的关系;通过LipofectamineTM 2000将miRNA模拟物和质粒分别或者联合转染细胞;TargetScanHuman预测及双荧光素酶报告验证miR-101-3p对STC-1的靶向调控关系;划痕实验、Transwell小室实验、Matrigel体外成管实验及Westernblot检测验证miR-101-3p靶向STC-1基因对癌细胞的侵袭转移和血管生成的影响及可能的机制,并通过裸鼠致瘤实验检测移植瘤的发展。结果 胃癌组织中STC-1的表达水平高于正常组织。与正常胃组织和GES-1细胞相比,胃癌组织和BGC-823细胞中miR-101-3p下调,STC-1 mRNA上调,且miR-101-3p水平与STC-1水平负相关,miR-101-3p水平与肿瘤分化程度、TNM分期、淋巴结转移显著相关（P<0.05）;过表达miR-101-3p可抑制STC-1的表达,下调p-PI3K/PI3K、p-AKT/AKT、MMP-2、MMP-9、VEGF、Ang2水平,抑制肿瘤细胞的侵袭转移和血管生成,降低移植瘤体积和重量（P<0.05）。结论 miR-101-3p在胃癌中表达下调,能够靶向STC-1基因调控PI3K/AKT信号通路抑制胃癌细胞BGC-823的侵袭转移和血管生成及体内移植瘤的进展。     

miR-101-3p sensitizes lung adenocarcinoma cells to irradiation via targeting BIRC5

Lung adenocarcinoma (LUAD) has been considered as the most common cause of cancer-associated mortality. Radiotherapy resistance is one of the main reasons for LUAD treatment failure. The microRNA (miR)-101-3p has been previously reported to function as a tumor suppressor in several types of cancer, including LUAD. The present study aimed to explore the role and mechanism of miR-101-3p on radioresistance of lung adenocarcinoma cells through bioinformatics analysis and biological experiments. Based on the analysis of Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) data, it was demonstrated that the expression of miR-101-3p was low in LUAD tissues compared with normal lung tissues and was associated with poor prognosis of patients with LUAD. The results of the CCK-8 assay, colony formation assay, immunofluorescence staining, caspase-3 activity assay and western blotting demonstrated that miR-101-3p overexpression sensitized LUAD cells to ionizing radiation by decreasing the abilities of LUAD cell proliferation, colony formation, DNA damage repair and increasing caspase-3 activity and apoptosis of LUAD cells following ionizing radiation. Furthermore, according to bioinformatics analysis and luciferase assay, baculoviral IAP repeat containing 5 (BIRC5) was identified as a direct target of miR-101-3p. Increased BIRC5 expression reversed the miR-101-3p-mediated increase in LUAD cell radiotherapy sensitivity. Taken together, the results of the present study demonstrated that miR-101-3p may be considered as a potential target that can enhance LUAD cell sensitivity to radiotherapy, which may provide a new strategy to improve therapy in patients with LUAD.     

液态活检miR-509-5p/miR-124比值诊断膀胱癌的临床应用

背景与目的:液态活检是一种从血液、尿液等非实体生物组织中取样分析,主要用于恶性肿瘤诊断、监测及判断预后的方法.该研究通过优化尿液miRNA的提取方法,建立标准化的液态活检手段,筛选并验证膀胱癌患者尿液miRNA标志物以阐明其临床应用价值.方法:收集2014年1月—2015年9月膀胱癌患者及健康对照者的尿液,应用miRNA表达谱芯片进行筛选,在78例膀胱癌患者及23例健康对照者的晨尿中进行实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)验证,分析液态活检的miRNA标志物与膀胱癌临床病理的关系及其诊断价值.结果:通过对6例膀胱癌及6例健康对照者的尿液miRNA表达谱进行分析,筛选出了10个miRNA有差异表达,结合既往文献报道的膀胱癌尿液miRNA标志物,我们挑选了20个miRNA进行RTFQ-PCR验证,发现miR-509-5p/miR-124比值在膀胱癌患者尿液中表达水平较健康对照者高,差异有统计学意义(P<0.0001).尿液miR-509-5p/miR-124比值随着患者肿瘤分期和肿瘤分级的上升而升高(P=0.003).当界值定在0.41时,miR-509-5p/miR-124比值对膀胱癌的诊断灵敏度为73.1%,特异度为82.6%,曲线下面积(area under the curve,AUC)达到0.864,比尿液脱落细胞学的诊断价值高(P=0.0002).结论:该研究优化了尿液miRNA的提取方法,建立了标准化的液态活检尿液miRNA标志物的检测手段,发现miR-509-5p/miR-124比值可能是膀胱癌理想的诊断标志物.     

Downregulated miR-367-3p,miR-548aq-5p,and miR-4710 in Human Whole Blood: Potential Biomarkers for Breast Cancer With Axillary Lymph Node Metastasis

BackgroundIncreasing studies have shown that microRNAs (miRNAs) have great diagnostic value in cancer. Axillary lymph node metastasis (ALNM) is closely related to the prognosis of breast cancer. However, it remains unknown whether miRNAs in whole blood could be promising biomarkers in breast cancer ALNM.MethodsAn miRNA microarray was used to screen potential differentially expressed miRNA candidates in whole blood of three breast cancer patients with ALNM and three without ALNM. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect candidate differentially expressed miRNAs in the whole blood of 109 breast cancer patients. Furthermore, bioinformatics analysis was carried to predict the potential targets and enriched pathway of miRNAs.ResultsQRT-PCR validated the fact that miR-367-3p, miR-548aq-5p and miR-4710 are downregulated in breast cancer with ALNM compared to it without ALNM. Receiver operating characteristic (ROC) curve analysis revealed that miR-367-3p, miR-548aq-5p and miR-4710 have good diagnostic values. Notably, the three-miRNA signature showed better predictive value, with an area under ROC curve (AUC) of 0.7414. Bioinformatics analysis revealed that the miRNAs could participate in a complex network and thus be involved in cancer-related pathways.ConclusionsOur findings support the potential of miR-367-3p, miR-548aq-5p and miR-4710 and the three-miRNA signature as biomarkers for breast cancer with ALNM.