Nucleic Acids Research, 1993, Vol. 21, No. 16 3809-3819
© 1993
COMPUTATIONAL BIOLOGY |
Problems related to the interpretation of autoradiographic data on gene expression using common constitutive transcripts as controls
Institut d'Oncologie Cellulaire et Moléculaire Humaine 129 rue de Stalingrad, F-93000 Bobigny, France
Received March 30, 1993. Revised June 28, 1993. Accepted June 28, 1993.
The 28S rRNA, a ribosomal RNA, and the ACTB and GAPD mRNAs, coding respectively for ß3-actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), are frequently presented as controls of modulated gene expression. These transcripts were quantified by replicate slot-blot autoradiography and image analysis in mammary epithelial cells and fibroblasts from breast tissues. Each cell-type group comprised strains with different pathological backgrounds, growth rates, antigenic phenotypes and culture histories. The effects of a differentiating agent (cholera toxin) and/or a tumor promoter (12-O-tetradecanoyl-phorboH3-acetate) were also examined. Despite the impression that visual examination of autoradiographs might create, image analysis suggests that 28S rRNA, ACTB and GAPD are substantially and independently influenced by the above biological factors and by the drugs. Therefore, these transcripts represent specifically regulated cellular activities and may not be taken as alternative indicators of the overall transcription rate or of the amount of material being examined. Instead, such nonspecific variation may be accurately measured and removed from quantitative data using a principal component function. A methodology that allows comparison of expression (or amplification) patterns between genes, between experiments or, even, between laboratories is presented with an example of quantification of transcripts related to cell-growth, differentiation, signaling and cancer.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  A. Zhang, L. Pastor, Q. Nguyen, Y. Luo, W. Yang, M. Flagella, R. Chavli, S. Bui, C. T. Nguyen, Z. Zheng, et al. Small Interfering RNA and Gene Expression Analysis Using a Multiplex Branched DNA Assay without RNA Purification J Biomol Screen, September 1, 2005; 10(6): 549 - 556. [Abstract] [PDF]
|
|
|
|
 |
|
 S A Bustin, V Benes, T Nolan, and M W Pfaffl Quantitative real-time RT-PCR - a perspective J. Mol. Endocrinol., June 1, 2005; 34(3): 597 - 601. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. K. H. Chang, S. M. Bandiera, and J. Chen Response to Letter to the Editor Drug Metab. Dispos., May 1, 2003; 31(5): 686 - 686. [Full Text] [PDF]
|
|
|
|
 |
|
 D. S. Hunter, M. Klotzbucher, H. Kugoh, S.-L. Cai, J. P. Mullen, G. Manfioletti, U. Fuhrman, and C. L. Walker Aberrant Expression of HMGA2 in Uterine Leiomyoma Associated with Loss of TSC2 Tumor Suppressor Gene Function Cancer Res., July 1, 2002; 62(13): 3766 - 3772. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. J. Landesberg, R. Ramalingam, K. Lee, T. K. Rosengart, and R. G. Crystal Upregulation of transcription factors in lung in the early phase of postpneumonectomy lung growth Am J Physiol Lung Cell Mol Physiol, November 1, 2001; 281(5): L1138 - L1149. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. M. Hellmann, W. R. Fields, and D. J. Doolittle Gene Expression Profiling of Cultured Human Bronchial Epithelial and Lung Carcinoma Cells Toxicol. Sci., May 1, 2001; 61(1): 154 - 163. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Z. Li, R. O. Stuart, J. Qiao, A. Pavlova, K. T. Bush, M. Pohl, H. Sakurai, and S. K. Nigam A role for Timeless in epithelial morphogenesis during kidney development PNAS, August 29, 2000; 97(18): 10038 - 10043. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Hatada, W. Kuziel, O. Smithies, and N. Maeda The Influence of Chromosomal Location on the Expression of Two Transgenes in Mice J. Biol. Chem., January 8, 1999; 274(2): 948 - 955. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. B. White, J. A. Eisen, T. L. Kasten, and R. D. Fernald Second gene for gonadotropin-releasing hormone in humans PNAS, January 6, 1998; 95(1): 305 - 309. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. M. Cale, D. S. Millican, H. Itoh, R. R. Magness, and I. M. Bird Pregnancy Induces an Increase in the Expression of Glyceraldehyde-3-Phosphate Dehydrogenase in Uterine Artery Endothelial Cells Reproductive Sciences, November 1, 1997; 4(6): 284 - 292. [Abstract] [PDF]
|
|
|
|
 |
|
 C. L. Cooper, G. Brady, F. Bilia, N. N. Iscove, and P. J. Quesenberry Expression of the Id Family Helix-Loop-Helix Regulators During Growth and Development in the Hematopoietic System Blood, May 1, 1997; 89(9): 3155 - 3165. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Macalma, J. Otte, M. E. Hensler, S. M. Bockholt, H. A. Louis, M. Kalff-Suske, K.-H. Grzeschik, D. von der Ahe, and M. C. Beckerle Molecular Characterization of Human Zyxin J. Biol. Chem., December 6, 1996; 271(49): 31470 - 31478. [Abstract] [Full Text] [PDF]
|
|