[Autocrine stimulation of receptor-tyrosine kinases (RTK) in human tumor cell lines in vitro: therapeutic implications].
OBJECTIVE
The VEGF/VEGFR system is known to play an important role in the development of new blood vessels during tumor formation. There is evidence that VEGFRs are not only present on endothelial cells but also on tumor cells. Since VEGF is able to induce proliferation and migration via VEGFR-2 we have studied the expression of VEGFRs and related receptor tyrosine kinases (RTKs) in different tumor cell lines and the effect of growth factor stimulation.


METHODS
RTK expression was investigated in 5 different human tumor cell lines on protein and mRNA levels. Tumor cell lines were exposed to growth factors such as VEGF and the phosphorylation of downstream molecules involved in proliferation, migration and apoptosis were assessed. Under comparable conditions proliferation and migration essays were performed. Endogenous production of VEGF and PDGF by the tumor cells was measured by ELISA of cell culture supernatants.


RESULTS
Most tested cell lines expressed all known VEGFR's, PDGFR-beta on protein and mRNA levels to a varying extent. 3 out of 5 cell lines could be stimulated after addition of VEGF reflected by an increased phosphorylation of MAPK, AKT/PKB and to a lesser extent of p38. This was underlined by an increased cell number and reduced number of apoptotic cells. After stimulation with PDGF-BB a stronger induction of MAPK and AKT/PKB phosphorylation than for VEGF could be seen. In contrast, no effect on tumor cell migration was detectable in all examined cell lines. The investigation of cell culture supernatants revealed that most cell lines do not produce VEGF or PDGF.


CONCLUSIONS
Tumor cell lines express RTKs and the receptor is stimulable after addition of growth factors such as VEGF. Thus, secretion of groth factors in the tumor microenvironment is not only able to stimulate proliferation and survival of endothelial cells but also tumor cells themselve. One cell line displayed high levels of endogenous VEGF which could explain the lack of an increased cell number after addition of VEGF. It remains obscure why another cell line could not be stimulated although receptors were present at the cellular surface. Further investigations should prove that RTK's could be influenced by therapeutic drugs in order to suppress cell proliferation and migration and induce apoptosis in tumor cell lines.
Gaumann
A
A
Max-Planck-Institut für physiologische und klinische Forschung, Abteilung molekulare Zellbiologie, Bad Nauheim.
Groot
M
M
Drexler
H C
HC
Breier
G
G
D045744:Cell Line, Tumor; D004789:Enzyme Activation; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000235:genetics
D012333:RNA, Messenger; D012334:RNA, Neoplasm; D042461:Vascular Endothelial Growth Factor A; D020794:Receptor Protein-Tyrosine Kinases; D040281:Vascular Endothelial Growth Factor Receptor-1; D040301:Vascular Endothelial Growth Factor Receptor-2; D040321:Vascular Endothelial Growth Factor Receptor-3