Comparison of miRNA expression patterns using total RNA extracted from matched samples of formalin-fixed paraffin-embedded (FFPE) cells and snap frozen cells

被引:272
作者
Li, Jinghuan
Smyth, Paul
Flavin, Richard
Cahill, Susanne
Denning, Karen
Aherne, Sinead
Guenther, Simone M.
O'Leary, John J.
Sheils, Orla [1 ]
机构
[1] Univ Dublin Trinity Coll, Dept Histopathol, Dublin 2, Ireland
[2] Appl Biosyst Inc, Foster City, CA USA
关键词
D O I
10.1186/1472-6750-7-36
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Background: Archival formalin-fixed paraffin-embedded (FFPE) tissues have limited utility in applications involving analysis of gene expression due to mRNA degradation and modification during fixation and processing. This study analyzed 160 miRNAs in paired snap frozen and FFPE cells to investigate if miRNAs may be successfully detected in archival specimens. Results: Our results show that miRNA extracted from FFPE blocks was successfully amplified using Q-RT-PCR. The levels of expression of miRNA detected in total RNA extracted from FFPE were higher than that extracted from snap frozen cells when the quantity of total RNA was identical. This phenomenon is most likely explained by the fact that larger numbers of FFPE cells were required to generate equivalent quantities of total RNA than their snap frozen counterparts. Conclusion: We hypothesise that methylol cross-links between RNA and protein which occur during tissue processing inhibit the yield of total RNA. However, small RNA molecules appear to be less affected by this process and are recovered more easily in the extraction process. In general miRNAs demonstrated reliable expression levels in FFPE compared with snap frozen paired samples, suggesting these molecules might prove to be robust targets amenable to detection in archival material in the molecular pathology setting.
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页数:6
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共 16 条
[1]   Towards quantitative mRNA analysis in paraffin-embedded tissues using real-time reverse transcriptase-polymerase chain reaction - A methodological study on lymph nodes from melanoma patients [J].
Abrahamsen, HN ;
Steiniche, T ;
Nexo, E ;
Hamilton-Dutoit, SJ ;
Sorensen, BS .
JOURNAL OF MOLECULAR DIAGNOSTICS, 2003, 5 (01) :34-41
[2]   Reliable gene expression measurements from degraded RNA by quantitative real-time PCR depend on short amplicons and a proper normalization [J].
Antonov, J ;
Goldstein, DR ;
Oberli, A ;
Baltzer, A ;
Pirotta, M ;
Fleischmann, A ;
Altermatt, HJ ;
Jaggi, R .
LABORATORY INVESTIGATION, 2005, 85 (08) :1040-1050
[3]   Phylogenetic shadowing and computational identification of human microRNA genes [J].
Berezikov, E ;
Guryev, V ;
van de Belt, J ;
Wienholds, E ;
Plasterk, RHA ;
Cuppen, E .
CELL, 2005, 120 (01) :21-24
[4]   Effect of ret/PTC 1 rearrangement on transcription and post-transcriptional regulation in a papillary thyroid carcinoma model [J].
Cahill, Susanne ;
Smyth, Paul ;
Finn, Stephen P. ;
Denning, Karen ;
Flavin, Richard ;
O'Regan, Esther M. ;
Li, Jinghuan ;
Potratz, Astrid ;
Guenther, Simone M. ;
Henfrey, Richard ;
O'Leary, John J. ;
Sheils, Orla .
MOLECULAR CANCER, 2006, 5 (1)
[5]   Real-time quantification of microRNAs by stem-loop RT-PCR [J].
Chen, CF ;
Ridzon, DA ;
Broomer, AJ ;
Zhou, ZH ;
Lee, DH ;
Nguyen, JT ;
Barbisin, M ;
Xu, NL ;
Mahuvakar, VR ;
Andersen, MR ;
Lao, KQ ;
Livak, KJ ;
Guegler, KJ .
NUCLEIC ACIDS RESEARCH, 2005, 33 (20) :e179.1-e179.9
[6]   Measurement of gene expression in archival paraffin-embedded tissues - Development and performance of a 92-gene reverse transcriptase-polymerase chain reaction assay [J].
Cronin, M ;
Pho, M ;
Dutta, D ;
Stephans, JC ;
Shak, S ;
Kiefer, MC ;
Esteban, JM ;
Baker, JB .
AMERICAN JOURNAL OF PATHOLOGY, 2004, 164 (01) :35-42
[7]   Quantitative mRNA expression analysis from formalin-fixed, paraffin-embedded tissues using 5′ nuclease quantitative reverse transcription-polymerase chain reaction [J].
Godfrey, TE ;
Kim, SH ;
Chavira, M ;
Ruff, DW ;
Warren, RS ;
Gray, JW ;
Jensen, RH .
JOURNAL OF MOLECULAR DIAGNOSTICS, 2000, 2 (02) :84-91
[8]   miRBase: microRNA sequences, targets and gene nomenclature [J].
Griffiths-Jones, Sam ;
Grocock, Russell J. ;
van Dongen, Stijn ;
Bateman, Alex ;
Enright, Anton J. .
NUCLEIC ACIDS RESEARCH, 2006, 34 :D140-D144
[9]   The role of microRNA genes in papillary thyroid carcinoma [J].
He, HL ;
Jazdzewski, K ;
Li, W ;
Liyanarachchi, S ;
Nagy, R ;
Volinia, S ;
Calin, GA ;
Liu, CG ;
Franssila, K ;
Suster, S ;
Kloos, RT ;
Croce, CM ;
de la Chapelle, A .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2005, 102 (52) :19075-19080
[10]   Real-time quantitative RT-PCR shows variable, assay-dependent sensitivity to formalin fixation: Implications for direct comparison of transcript levels in paraffin-embedded tissues [J].
Koch, Ina ;
Slotta-Huspenina, Julia ;
Hollweckt, Regina ;
Anastasov, Natasa ;
Hofler, Heinz ;
Quintanilla-Martinez, Leticia ;
Fend, Falko .
DIAGNOSTIC MOLECULAR PATHOLOGY, 2006, 15 (03) :149-156