PROBLEMS RELATED TO THE INTERPRETATION OF AUTORADIOGRAPHIC DATA ON GENE-EXPRESSION USING COMMON CONSTITUTIVE TRANSCRIPTS AS CONTROLS

被引:144
作者
SPANAKIS, E
机构
[1] Institut d'Oncologie Cellulaire et Moléculaire Humaine, F-93000 Bobigny
关键词
D O I
10.1093/nar/21.16.3809
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The 28S rRNA, a ribosomal RNA, and the ACTB and GAPD mRNAs, coding respectively for beta-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-phorbol-13-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 non-specific 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.
引用
收藏
页码:3809 / 3819
页数:11
相关论文
共 22 条
[1]  
AITKEN SC, 1985, CANCER RES, V45, P1611
[2]   A NOVEL METALLOPROTEINASE GENE SPECIFICALLY EXPRESSED IN STROMAL CELLS OF BREAST CARCINOMAS [J].
BASSET, P ;
BELLOCQ, JP ;
WOLF, C ;
STOLL, I ;
HUTIN, P ;
LIMACHER, JM ;
PODHAJCER, OL ;
CHENARD, MP ;
RIO, MC ;
CHAMBON, P .
NATURE, 1990, 348 (6303) :699-704
[3]  
BEAUPAIN R, 1993, IN VITRO CELL DEV-AN, V29, P100
[4]   FETAL MYOFIBROBLAST-LIKE CELLS ISOLATED FROM POSTRADIATION FIBROSIS IN HUMAN BREAST-CANCER [J].
BROUTYBOYE, D ;
RAUX, H ;
AZZARONE, B ;
TAMBOISE, A ;
TAMBOISE, E ;
BERANGER, S ;
MAGNIEN, V ;
PIHAN, I ;
ZARDI, L ;
ISRAEL, L .
INTERNATIONAL JOURNAL OF CANCER, 1991, 47 (05) :697-702
[5]  
BROUTYBOYE D, UNPUB
[6]  
CHANG FH, 1989, J BIOL CHEM, V264, P5352
[7]   GENOMIC SEQUENCING [J].
CHURCH, GM ;
GILBERT, W .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1984, 81 (07) :1991-1995
[8]  
COSSEL D, 1978, P NATL ACAD SCI USA, V75, P2669
[9]  
Davis L. G., 1986, BASIC METHODS MOL BI
[10]  
GAMBY C, 1992, CANCER DETECT PREV, V16, P71