Quantitation of vascular endothelial growth factor mRNA levels in human breast tumors and metastatic lymph nodes.
In situ hybridization analysis provides a means to qualitatively study the heterogeneity of primary tumors and metastases based on the types of genes transcribed. In this study, we have tested some parameters for quantitative analysis of in situ hybridizations with paraffin-embedded human breast tumors and measured mRNA levels for the angiogenic protein, vascular endothelial growth factor (VEGF). VEGF mRNAs were highly tumor specific, with the highest levels near necrotic regions within the tissues (0.1 to 2.7 dpm/mm2). Normal cells within the tissue sections did not have detectable levels of VEGF mRNA. For comparison, tumor levels of c-myc (4 to 46 dpm/mm2) and glyceraldehyde-3-phosphate dehydrogenase mRNAs (48 to 214 dpm/mm2) were measured. The mRNAs for both of these genes were more broadly expressed across the tissue sections. The hybridization pattern for VEGF mRNAs was consistent with hypoxia-induced VEGF mRNA steady-state levels and supports the hypothesis that oxidative stress regulates VEGF expression in breast tumors.
Wright
P S
PS
Hoechst Marion Roussel, Cincinnati, Ohio 45215, USA.
Loudy
D E
DE
Cross-Doersen
D E
DE
Montgomery
L R
LR
Sprinkle-Cavallo
J
J
Miller
J A
JA
Distler
C M
CM
Lower
E E
EE
Woessner
R D
RD
D014408:Biomarkers, Tumor; D001943:Breast Neoplasms / Q000737:chemistry* / Q000473:pathology; D017931:DNA Primers / Q000737:chemistry; D016228:Endothelial Growth Factors / Q000032:analysis* / Q000235:genetics; D005260:Female; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000032:analysis; D006801:Humans; D017403:In Situ Hybridization; D008198:Lymph Nodes / Q000737:chemistry* / Q000473:pathology; D008207:Lymphatic Metastasis; D008222:Lymphokines / Q000032:analysis* / Q000235:genetics; D016271:Proto-Oncogene Proteins c-myc / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis*; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D014408:Biomarkers, Tumor; D017931:DNA Primers; D016228:Endothelial Growth Factors; D008222:Lymphokines; D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases