Statistical modeling for selecting housekeeper genes

There is a need for statistical methods to identify genes that have minimal variation in expression across a variety of experimental conditions. These 'housekeeper' genes are widely employed as controls for quantification of test genes using gel analysis and real-time RT-PCR. Using real-time quantitative RT-PCR, we analyzed 80 primary breast tumors for variation in expression of six putative housekeeper genes (MRPL19 (mitochondrial ribosomal protein L19), PSMC4 (proteasome (prosome, macropain) 26S subunit, ATPase, 4), SF3A1 (splicing factor 3a, subunit 1, 120 kDa), PUM1 (pumilio homolog 1 (Drosophila)), ACTB (actin, beta) and GAPD (glyceraldehyde-3-phosphate dehydrogenase)). We present appropriate models for selecting the best housekeepers to normalize quantitative data within a given tissue type (for example, breast cancer) and across different types of tissue samples.

[1]  Carl T. Wittwer,et al.  Rapid Cycle Real-Time PCR — Methods and Applications , 2002, Springer Berlin Heidelberg.

[2]  W. Taylor,et al.  Comparison of glyceraldehyde-3-phosphate dehydrogenase and 28S-ribosomal RNA gene expression as RNA loading controls for northern blot analysis of cell lines of varying malignant potential. , 1994, Analytical biochemistry.

[3]  D. Bates,et al.  Mixed-Effects Models in S and S-PLUS , 2001 .

[4]  E. Spanakis,et al.  Problems related to the interpretation of autoradiographic data on gene expression using common constitutive transcripts as controls. , 1993, Nucleic acids research.

[5]  P. J. Higgins,et al.  Control selection for RNA quantitation. , 2000, BioTechniques.

[6]  G. Schwarz Estimating the Dimension of a Model , 1978 .

[7]  Charles M. Perou,et al.  Tumor classification using gene expression patterns from DNA microarrays , 2000 .

[8]  V. Carey,et al.  Mixed-Effects Models in S and S-Plus , 2001 .

[9]  R. Rasmussen Quantification on the LightCycler , 2001 .

[10]  A. Eggert,et al.  Relative quantitative RT-PCR protocol for TrkB expression in neuroblastoma using GAPD as an internal control. , 2000, BioTechniques.

[11]  S. Akilesh,et al.  Customized molecular phenotyping by quantitative gene expression and pattern recognition analysis. , 2003, Genome research.

[12]  D. Lockhart,et al.  Analysis of gene expression profiles in normal and neoplastic ovarian tissue samples identifies candidate molecular markers of epithelial ovarian cancer. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Paul S Mischel,et al.  Molecular Analysis of Glioblastoma: Pathway Profiling and Its Implications for Patient Therapy , 2003, Cancer biology & therapy.

[14]  F. Pichaud,et al.  Effects of Prostaglandins on Human Hematopoietic Osteoclast Precursors. , 1997, Endocrinology.

[15]  L J Kricka,et al.  Evaluation of DNA fragment sizing and quantification by the agilent 2100 bioanalyzer. , 2000, Clinical chemistry.

[16]  S. Dhanasekaran,et al.  Delineation of prognostic biomarkers in prostate cancer , 2001, Nature.

[17]  Yudong D. He,et al.  A Gene-Expression Signature as a Predictor of Survival in Breast Cancer , 2002 .

[18]  F. Speleman,et al.  Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes , 2002, Genome Biology.

[19]  P. Bernard,et al.  Profiling Breast Cancer Using Real-Time Quantitative PCR , 2004 .

[20]  J. SantaLucia,et al.  A unified view of polymer, dumbbell, and oligonucleotide DNA nearest-neighbor thermodynamics. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[21]  R. Yolken,et al.  Methods to optimize the generation of cDNA from postmortem human brain tissue. , 2003, Brain research. Brain research protocols.

[22]  Christian A. Rees,et al.  Molecular portraits of human breast tumours , 2000, Nature.

[23]  T. Grogan,et al.  Discrepancies in clinical laboratory testing of eligibility for trastuzumab therapy: apparent immunohistochemical false-positives do not get the message. , 2001, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[24]  Ronald W. Davis,et al.  Quantitative Monitoring of Gene Expression Patterns with a Complementary DNA Microarray , 1995, Science.