RT-PCR method to quantify vascular endothelial growth factor expression.
The vascular endothelial growth factor (VEGF) is implicated in the progression of cancers. Its expression is well correlated with tumor growth and metastases. The availability of a rapid and sensitive method to detect the amounts of VEGF mRNA in biological samples of limited size, very small biopsies, or samples containing relatively few cells could provide an interesting prognostic tool for clinicians. We have developed an RT-PCR method that allows us to detect the VEGF mRNA from as little as 3 micrograms total mRNA. We have also shown that this protocol can be generalized to all cell lines tested. This method constitutes a very potent tool for the analysis of VEGF mRNA expression in different contexts.
Bonino
F
F
CNRS UMR 6543, Centre Antoine Lacassagne, 33 Avenue de Valombrose, 06189 Nice, France.
Milanini
J
J
Pouysségur
J
J
Pagès
G
G
D000230:Adenocarcinoma / Q000473:pathology; D000818:Animals; D001667:Binding, Competitive; D001706:Biopsy; D002460:Cell Line / Q000187:drug effects / Q000378:metabolism; D003110:Colonic Neoplasms / Q000473:pathology; D006224:Cricetinae; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D016228:Endothelial Growth Factors / Q000096:biosynthesis* / Q000235:genetics; D004958:Estradiol / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008222:Lymphokines / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D008832:Microchemistry / Q000379:methods; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D009389:Neovascularization, Pathologic / Q000235:genetics; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000235:genetics; D012326:RNA Splicing; D012333:RNA, Messenger / Q000032:analysis* / Q000096:biosynthesis / Q000235:genetics; D012334:RNA, Neoplasm / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D012015:Reference Standards; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017384:Sequence Deletion; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D017931:DNA Primers; D018076:DNA, Complementary; D016228:Endothelial Growth Factors; D008222:Lymphokines; D009363:Neoplasm Proteins; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D004958:Estradiol