The GLI (GLI1/GLI2) transcription factors have been implicated in the development
and progression of prostate cancer although our understanding of how they
actually contribute to the biology of these common tumours is limited. We
observed that GLI reporter activity was higher in normal (PNT-2) and
tumourigenic (DU145 and PC-3) androgen-independent cells compared to
androgen-dependent LNCaP prostate cancer cells and, accordingly, GLI mRNA levels
were also elevated. Ectopic expression of GLI1 or the constitutively active
ΔNGLI2 mutant induced a distinct cobblestone-like morphology in LNCaP cells
that, regarding the former, correlated with increased GLI2 as well as expression
of the basal/stem-like markers CD44, β1-integrin, ΔNp63 and BMI1, and
decreased expression of the luminal marker AR (androgen receptor). LNCaP-GLI1
cells were viable in the presence of the AR inhibitor bicalutamide and gene
expression profiling revealed that the transcriptome of LNCaP-GLI1 cells was
significantly closer to DU145 and PC-3 cells than to control LNCaP-pBP (empty
vector) cells, as well as identifying LCN2/NGAL as a highly induced transcript
which is associated with hormone independence in breast and prostate cancer.
Functionally, LNCaP-GLI1 cells displayed greater clonal growth and were more
invasive than control cells but they did not form colonies in soft agar or
prostaspheres in suspension suggesting that they do not possess inherent stem
cell properties. Moreover, targeted suppression of GLI1 or GLI2 with siRNA did
not reverse the transformed phenotype of LNCaP-GLI1 cells nor did double
GLI1/GLI2 knockdowns activate AR expression in DU145 or PC-3 cells. As such,
early targeting of the GLI oncoproteins may hinder progression to a hormone
independent state but a more detailed understanding of the mechanisms that
maintain this phenotype is required to determine if their inhibition will
enhance the efficacy of anti-hormonal therapy through the induction of a luminal
phenotype and increased dependency upon AR function