52 research outputs found
Reference miRNAs for miRNAome Analysis of Urothelial Carcinomas
Background/Objective: Reverse transcription quantitative real-time PCR (RT-qPCR) is widely used in microRNA (miRNA) expression studies on cancer. To compensate for the analytical variability produced by the multiple steps of the method, relative quantification of the measured miRNAs is required, which is based on normalization to endogenous reference genes. No study has been performed so far on reference miRNAs for normalization of miRNA expression in urothelial carcinoma. The aim of this study was to identify suitable reference miRNAs for miRNA expression studies by RT-qPCR in urothelial carcinoma. Methods: Candidate reference miRNAs were selected from 24 urothelial carcinoma and normal bladder tissue samples by miRNA microarrays. The usefulness of these candidate reference miRNAs together with the commonly for normalization purposes used small nuclear RNAs RNU6B, RNU48, and Z30 were thereafter validated by RT-qPCR in 58 tissue samples and analyzed by the algorithms geNorm, NormFinder, and BestKeeper. Principal Findings: Based on the miRNA microarray data, a total of 16 miRNAs were identified as putative reference genes. After validation by RT-qPCR, miR-101, miR-125a-5p, miR-148b, miR-151-5p, miR-181a, miR-181b, miR-29c, miR-324-3p, miR-424, miR-874, RNU6B, RNU48, and Z30 were used for geNorm, NormFinder, and BestKeeper analyses that gave different combinations of recommended reference genes for normalization. Conclusions: The present study provided the first systematic analysis for identifying suitable reference miRNAs for miRNA expression studies of urothelial carcinoma by RT-qPCR. Different combinations of reference genes resulted in reliable expression data for both strongly and less strongly altered miRNAs. Notably, RNU6B, which is the most frequently used reference gene for miRNA studies, gave inaccurate normalization. The combination of four (miR-101, miR-125a-5p, miR-148b, and miR-151-5p) or three (miR-148b, miR-181b, and miR-874,) reference miRNAs is recommended for normalization
Micromorfologische studie van mortel-, pleister- en vloerbetonfragmenten in de Civitas Menapiorum
Petrografisch onderzoek van de natuursteen uit de H.-Kruiskerk te Vrasene (periode II: 1153-1183
De mortels uit de H.-Kruiskerk te Vrasene (periode II en III: 1153-1183 en 1350-1400). Chemisch en micromorfologisch onderzoek
Pedosedimentary and climatological Record of the Loess-Paleosol-Sequence during the last interglacial and early glacial age in Tadjikistan
Pedosedimentary evolution of the last interglacial and early glacial sequence in the European loess belt from Belgium to central Russia
Remicourt (Hesbaye): a pedostratigraphical record for the Last Interglacial and Early Glacial in Belgium: a testcase for NW Europe
The Chronostratigraphic Distribution and Chronological Interpretation of the Wallertheim Sequence: Comparisons with the Loess Paleosols of Tönchesberg and Middle Belgium
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