Background.Carcinoma of the prostate (CaP) is an increasing healthcare problem.

Given the poor

outcome of patients with castration-resistant prostate cancer (CRPC), new strategies are needed to
improve the current therapeutic regimens, slowdown the insurgence of hormone refractory disease
and/or develop novel treatments. Emerging evidence demonstrates a key role for the PI3K-AKTmTOR signaling axis in the development and maintenance of CRPC. This pathway, which is
deregulated in the majority of advanced prostate cancers, serves as a critical nexus for the
integration of growth signals with downstream cellular processes such as protein synthesis,
proliferation, survival, metabolism and differentiation, thus providing mechanisms for cancer cells
to overcome the stress associated with androgen deprivation. In addition, aberrant function of
mTOR has been shown to be present in the proliferation of Cancer Stem cells (CSCs) or tumor
initiating cells.
Aims. To investigate the role of TORC1 and TORC2 in the proliferation and apoptosis induced by
hormone manipulation in androgen sensitive or castration resistant prostate cancer models. In
addition we intend to evaluate the possible potentiating effects of thedual TORC1/TORC2 inhibitor,
RES529.
Results.Here, we demonstrated that siRNA for raptor (TORC1 silencing) had no significant change
in the growth of 22rv1 cells whereas the siRNA for rictor (TORC2 silencing) induced a marked
inhibition of cell proliferation. TORC1 silencing increased the expression of AR and Ser473 Akt
whereas TORC2 marked reduced them. This suggests that TORC2 is a promising target for prostate
cancer, therefore, RES-529 a dual TORC1/TORC2 inhibitor was tested in comparison with
RAD001 in experiments of combination with androgen deprivation and AR inhibition both in vitro
and in vivo. in vivo we tested the combination effects of 5-reductase (Finasteride or Dutasteride),
CYP17 (Abiraterone) and AR (Bicalutamide) inhibitors. We observed that RES-529 was able to
synergize with hormone manipulations in 22rv1 cells. In chronic in vitro experiments, RES-529
seems to prevent the insurgence of CRPC disease. Lower effects were showed by Rapamycin
analog (RAD001) a specific TORC1 inhibitor. The importance of TORC2 in this phenomenon was
also demonstrated by using Metformin, a well-known Adenosine monophosphate-activated protein
kinase (AMPK) inhibitor that phosphorylates TORC2, thereby blocking its nuclear translocation.
Metformin administration reduces TORC2 phosphorylation status increasing the effects of
RAD001. Conclusions: Our results provide a rationale for combinatorial treatment with
conventional hormone therapy and mTOR inhibitors, including TORC1/TORC2 dissociative
compounds like RES-529. This may be a promising therapeutic approach for the treatment of both
hormone sensitive and CR diseases.