Measurement of mRNA levels in tumor xenografts with quantitative autoradiography and in situ hybridization.
In situ methodologies allow qualitative and semi-quantitative analysis of spatial gene expression in whole organisms or tissues. We have applied quantitative autoradiography to in situ hybridizations of sections from human breast tumor xenografts to measure mRNA levels for ornithine decarboxylase, estrogen receptor, transforming growth factor alpha, and glyceraldehyde-3-phosphate dehydrogenase. Comparisons of control and tamoxifen-treated animals show significant decreases in MCF-7 tumor estrogen receptor mRNA levels in the drug-treated animals. Combining quantitative autoradiography with in situ hybridization allows measurement of absolute rather than relative mRNA levels for genes of interest, and to monitor effector-induced changes in these mRNAs in vivo.
Wright
P S
PS
Marion Merrell Dow Research Institute, Cincinnati, Ohio 45215-6300, USA.
Cross-Doersen
D E
DE
Chmielewski
P A
PA
Bush
T L
TL
Bitonti
A J
AJ
Miller
J A
JA
D000818:Animals; D001345:Autoradiography / Q000379:methods*; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D002250:Carbon Radioisotopes; D002460:Cell Line; D015342:DNA Probes; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D009955:Ornithine Decarboxylase / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000032:analysis*; D011960:Receptors, Estrogen / Q000096:biosynthesis*; D013629:Tamoxifen / Q000494:pharmacology; D016211:Transforming Growth Factor alpha / Q000096:biosynthesis; D014183:Transplantation, Heterologous; D014316:Tritium; D014407:Tumor Cells, Cultured
D002250:Carbon Radioisotopes; D015342:DNA Probes; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D016211:Transforming Growth Factor alpha; D013629:Tamoxifen; D014316:Tritium; D009955:Ornithine Decarboxylase