Monitoring signal transduction in cancer: cDNA microarray for semiquantitative analysis.
This study targeted the development of a novel microarray tool to allow rapid determination of the expression levels of 58 different tyrosine kinase (tk) genes in small tumor samples. The goals were to define a reference probe for multi-sample comparison and to investigate the variability and reproducibility of the image acquisition and RT-PCR procedures. The small number of tk genes on our arrays enabled us to define a reference probe by artificially mixing all genes on the arrays. Such a probe provided contrast reference for comparative hybridization of control and sample DNA and enabled cross-comparison of more than two samples against one another. Comparison of signals generated from multiple scanning eliminated the concern of photo bleaching and scanner intrinsic noise. Tests performed with breast, thyroid, and prostate cancer samples yielded distinctive patterns and suggest the feasibility of our approach. Repeated experiments indicated reproducibility of such arrays. Up- or downregulated genes identified by this rapid screening are now being investigated with techniques such as in situ hybridization.
Hsieh
H B
HB
Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
Lersch
R A
RA
Callahan
D E
DE
Hayward
S
S
Wong
M
M
Clark
O H
OH
Weier
H U
HU
D001943:Breast Neoplasms / Q000378:metabolism*; D005260:Female; D006801:Humans; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods / Q000592:standards; D011471:Prostatic Neoplasms / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013964:Thyroid Neoplasms / Q000378:metabolism*
D011505:Protein-Tyrosine Kinases