Effect of an EGF-R selective tyrosine kinase inhibitor and an anti-androgen on LNCaP cells: identification of divergent growth regulatory pathways.
BACKGROUND
The effect of an EGF-R selective tyrosine kinase (EGF-RTK) quinazoline inhibitor ZM252868 was determined on the androgen-sensitive human prostatic tumour cell line LNCaP, which can also respond via the EGF-R-regulated growth pathway for cell proliferation. Potential interaction or 'cross-talk' between steroid and the growth factor mitogen-activated protein kinase (MAPK) signalling pathway was also investigated.


METHODS
The responses of LNCaP cells to various growth factors in the absence and presence of the EGF-RTK inhibitor and/or steroid and anti-androgen Casodex, was determined using cell population analysis. The effect of the inhibitor on the expression of androgen receptor, EGF-R and activated MAPK was assessed immunocytochemically and changes in the MAPK signalling cascade were also determined using Western blotting techniques.


RESULTS
The ZM252868 inhibitor had no effect on LNCaP basal growth. At 100 nM and 1 microM concentrations, the inhibitor reduced the marked EGF- and TGF-alpha-stimulated LNCaP cell growth by 60% and to basal levels, respectively. Both bFGF- and 5alpha-DHT-stimulated growth were unaffected in this concentration range. The inhibitor (1 microM) decreased the expression of immunoreactive EGF-R but had no effect on androgen receptor levels. Activation of MAPK by EGF was noted, being down-regulated by the inhibitor at a concentration of 1 microM. MAPK was not activated by 5alpha-DHT. The anti-androgen Casodex reduced 5alpha-DHT-stimulated cell growth but had no effect on EGF-R mediated LNCaP growth or EGF-stimulated activated MAPK activity. Treatment with EGF and 5alpha-DHT in combination produced an additive effect on cell proliferation, with the anti-androgen and the EGF-RTK inhibitor only reducing the 5alpha-DHT- or EGF-stimulated portion of growth, respectively.


CONCLUSIONS
The study demonstrated the efficacy and selectivity of the ZM252868 inhibitor in inhibiting EGF-R mediated LNCaP cell growth. Additionally, no interaction between androgen and EGF-R mediated growth pathways was determined.
Jones
H E
HE
Tenovus Centre for Cancer Research, Welsh School of Pharmacy, Cardiff University, UK. joneshe1@cardiff.ac.uk
Barrow
D
D
Dutkowski
C M
CM
Goddard
L
L
Smith
C
C
Harper
M E
ME
Nicholson
R I
RI
D000726:Androgen Antagonists / Q000494:pharmacology*; D000813:Anilides / Q000494:pharmacology*; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects*; D004347:Drug Interactions; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D066246:ErbB Receptors / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D009570:Nitriles; D011471:Prostatic Neoplasms / Q000473:pathology*; D011799:Quinazolines / Q000494:pharmacology*; D015398:Signal Transduction; D014105:Tosyl Compounds; D014407:Tumor Cells, Cultured
D000726:Androgen Antagonists; D000813:Anilides; D004791:Enzyme Inhibitors; D009570:Nitriles; D011799:Quinazolines; D014105:Tosyl Compounds; C106485:ZM 252868; C053541:bicalutamide; D066246:ErbB Receptors