| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
INNOVATIVE METHODOLOGY
Canadian Institutes of Health Research Group in Functional Development and Physiopathology of the Digestive Tract, Département d'Anatomie et de Biologie Cellulaire, Faculté de Médecine, Université de Sherbrooke, Sherbrooke, Québec, Canada
Submitted 23 May 2005 ; accepted in final form 26 December 2005
As for other mRNA measurement methods, quantitative RT-PCR results need to be normalized relative to stably expressed genes. Widely used normalizing genes include 
-actin and glyceraldehyde-3-phosphate dehydrogenase. It has, however, become clear that these and other normalizing genes can display modulated patterns of expression across tissue types and during complex cellular processes such as cell differentiation and cancer progression. Our objective was to set the basis for identifying normalizing genes that displayed stable expression during enterocytic differentiation and between healthy tissue and adenocarcinomas of the human colon. We thus identified novel potential normalizing genes using previously generated cDNA microarray data and examined the alterations of expression of two of these genes as well as seven commonly used normalizing genes during the enterocytic differentiation process and between matched pairs of resection margins and primary carcinomas of the human colon using real-time RT-PCR. We found that ribosomal phosphoprotein P0 was particularly stable in all intestinal epithelial cell extracts, thereby representing a particularly robust housekeeping reference gene for the assessment of gene expression during the human enterocytic differentiation process. On the other hand, -2-microglobulin generated the best score as a normalizing gene for comparing human colon primary carcinomas with their corresponding normal mucosa of the resection margin, although others were found to represent acceptable alternatives. In conclusion, we identified and characterized specific normalizing genes that should significantly improve quantitative mRNA studies related to both the differentiation process of the human intestinal epithelium and adenocarcinomas of the human colon. This approach should also be useful to validate normalizing genes in other intestinal contexts.
mRNA analysis; intestinal cell differentiation; colon cancer cells
This article has been cited by other articles:
|
|
 |
|
  L. A. Emery, A. Tripathi, C. King, M. Kavanah, J. Mendez, M. D. Stone, A. de las Morenas, P. Sebastiani, and C. L. Rosenberg Early Dysregulation of Cell Adhesion and Extracellular Matrix Pathways in Breast Cancer Progression Am. J. Pathol., September 1, 2009; 175(3): 1292 - 1302. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Caporale, F. Arnaud, M. Mura, M. Golder, C. Murgia, and M. Palmarini The Signal Peptide of a Simple Retrovirus Envelope Functions as a Posttranscriptional Regulator of Viral Gene Expression J. Virol., May 1, 2009; 83(9): 4591 - 4604. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B Carvalho, C Postma, S Mongera, E Hopmans, S Diskin, M A van de Wiel, W van Criekinge, O Thas, A Matthai, M A Cuesta, et al. Multiple putative oncogenes at the chromosome 20q amplicon contribute to colorectal adenoma to carcinoma progression Gut, January 1, 2009; 58(1): 79 - 89. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L A Simms, J D Doecke, M D Walsh, N Huang, E V Fowler, and G L Radford-Smith Reduced {alpha}-defensin expression is associated with inflammation and not NOD2 mutation status in ileal Crohn's disease Gut, July 1, 2008; 57(7): 903 - 910. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Visit Other APS Journals Online |
