Aberrant microRNA expression in human cervical carcinomas.
ABSTRACT Because altered microRNAs (miRNAs) expression patterns have been observed in a variety of diseased tissues, miRNA expression was compared in human cervical cancer tissues relative to adjacent normal cervical tissues in the present study. Microarray chips with 924 probes were used to detect the expression of miRNAs in cervical cancer tissue and adjacent normal cervical tissue of 13 patients with cervical cancer (11 squamous cervical cancers, one cervical adenocarcinoma, and one cervical sarcoma), all of whom were infected with human papilloma virus (HPV) 16 and/or HPV18. Compared to the expression levels in normal cervical tissues, 18 miRNAs (1.9%) were significantly upregulated (increase of ≥2×), and 19 miRNAs (2.1%) were significantly downregulated (decrease of ≤0.5×) in cervical cancer tissues. miRNA expression was independent of lymph node involvement, vascular invasion, and pathological differentiation. Taken together, cervical cancer tissues have altered expression of miRNAs relative to adjacent normal tissues. Further studies are necessary to determine whether aberrant miRNA expression is related to the pathogenesis of cervical cancer.
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ABSTRACT: A number of recent studies have catalogued global gene expression patterns in a panel of normal, tumoral cervical tissues so that potential biomarkers can be identified. The qPCR has been one of the most widely used technologies for detecting these potential biomarkers. However, few studies have investigated a correct strategy for the normalization of data in qPCR assays for cervical tissues. The aim of this study was to validate reference genes in cervical tissues to ensure accurate quantification of mRNA and miRNA levels in cervical carcinogenesis. For this purpose, some issues for obtaining reliable qPCR data were evaluated such as the following: geNorm analysis with a set of samples which meet all of the cervical tissue conditions (Normal + CIN1 + CIN2 + CIN3 + Cancer); the use of individual Ct values versus pooled Ct values; and the use of a single (or multiple) reference genes to quantify mRNA and miRNA expression levels. Two different data sets were put on the geNorm to assess the expression stability of the candidate reference genes: the first dataset comprised the quantities of the individual Ct values; and the second dataset comprised the quantities of the pooled Ct values. Moreover, in this study, all the candidate reference genes were analyzed as a single “normalizer”. The normalization strategies were assessed by measuring p16INK4a and miR-203 transcripts in qPCR assays. We found that the use of pooled Ct values, can lead to a misinterpretation of the results, which suggests that the maintenance of inter-individual variability is a key factor in ensuring the reliability of the qPCR data. In addition, it should be stressed that a proper validation of the suitability of the reference genes is required for each experimental setting, since the indiscriminate use of a reference gene can also lead to discrepant results.PLoS ONE 11/2014; 9(11):e111021. · 3.53 Impact Factor
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ABSTRACT: microRNAs are a family of endogenous, short, non-coding RNAs that play critical roles in regulating gene expression for key cellular processes in normal and abnormal physiology. microRNA-7 is a 23 nucleotide miRNA whose expression is tightly regulated and restricted predominantly to the brain, spleen and pancreas. Reduced levels of miR-7 have been linked to the development of cancer and metastasis. As a tumor suppressor, miR-7 functions to co-ordinately downregulate a number of direct (e.g. the epidermal growth factor receptor) and indirect (e.g. phospho-Akt) growth promoting targets to decrease tumor growth in vitro and in vivo. In addition, miR-7 can increase the sensitivity of treatment-resistant cancer cells to therapeutics and inhibit metastasis. These data suggest that replacement of miR-7 ('miRNA replacement therapy') for specific human cancers could represent a new treatment approach.The international journal of biochemistry & cell biology. 06/2014;
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ABSTRACT: Small non-coding microRNAs (miRNAs) are involved in cancer development and progression, and serum profiles of cervical cancer patients may be useful for identifying novel miRNAs. We performed deep sequencing on serum pools of cervical cancer patients and healthy controls with 3 replicates and constructed a small RNA library. We used MIREAP to predict novel miRNAs and identified 2 putative novel miRNAs between serum pools of cervical cancer patients and healthy controls after filtering out pseudo-pre-miRNAs using Triplet-SVM analysis. The 2 putative novel miRNAs were validated by real time PCR and were significantly decreased in cervical cancer patients compared with healthy controls. One novel miRNA had an area under curve (AUC) of 0.921 (95% CI: 0.883, 0.959) with a sensitivity of 85.7% and a specificity of 88.2% when discriminating between cervical cancer patients and healthy controls. Our results suggest that characterizing serum profiles of cervical cancers by Solexa sequencing may be a good method for identifying novel miRNAs and that the validated novel miRNAs described here may be cervical cancer-associated biomarkers.Scientific reports. 01/2014; 4:6277.