PHLPPing the balance: restoration of protein kinase C in cancer

Protein kinase signalling, which transduces external messages to mediate cellular growth and metabolism, is frequently deregulated in human disease, and specifically in cancer. As such, there are 77 kinase inhibitors currently approved for the treatment of human disease by the FDA. Due to their historical association as the receptors for the tumour-promoting phorbol esters, PKC isozymes were initially targeted as oncogenes in cancer. However, a meta-analysis of clinical trials with PKC inhibitors in combination with chemotherapy revealed that these treatments were not advantageous, and instead resulted in poorer outcomes and greater adverse effects. More recent studies suggest that instead of inhibiting PKC, therapies should aim to restore PKC function in cancer: cancer-associated PKC mutations are generally loss-of-function and high PKC protein is protective in many cancers, including most notably KRAS-driven cancers. These recent findings have reframed PKC as having a tumour suppressive function. This review focusses on a potential new mechanism of restoring PKC function in cancer — through targeting of its negative regulator, the Ser/Thr protein phosphatase PHLPP. This phosphatase regulates PKC steady-state levels by regulating the phosphorylation of a key site, the hydrophobic motif, whose phosphorylation is necessary for the stability of the enzyme. We also consider whether the phosphorylation of the potent oncogene KRAS provides a mechanism by which high PKC expression may be protective in KRAS-driven human cancers.

miR-221 Augments TRAIL-Mediated Apoptosis in Prostate Cancer Cells by Inducing Endogenous TRAIL Expression and Targeting the Functional Repressors SOCS3 and PIK3R1

miR-221 is regarded as an oncogene in many malignancies, and miR-221-mediated resistance towards TRAIL was one of the first oncogenic roles shown for this small noncoding RNA. In contrast, miR-221 is downregulated in prostate cancer (PCa), thereby implying a tumour suppressive function. By using proliferation and apoptosis assays, we show a novel feature of miR-221 in PCa cells: instead of inducing TRAIL resistance, miR-221 sensitized cells towards TRAIL-induced proliferation inhibition and apoptosis induction. Partially responsible for this effect was the interferon-mediated gene signature, which among other things contained an endogenous overexpression of the TRAIL encoding gene TNFSF10. This TRAIL-friendly environment was provoked by downregulation of the established miR-221 target gene SOCS3. Moreover, we introduced PIK3R1 as a target gene of miR-221 in PCa cells. Proliferation assays showed that siRNA-mediated downregulation of SOCS3 and PIK3R1 mimicked the effect of miR-221 on TRAIL sensitivity. Finally, Western blotting experiments confirmed lower amounts of phospho-Akt after siRNA-mediated downregulation of PIK3R1 in PC3 cells. Our results further support the tumour suppressing role of miR-221 in PCa, since it sensitises PCa cells towards TRAIL by regulating the expression of the oncogenes SOCS3 and PIK3R1. Given the TRAIL-inhibiting effect of miR-221 in various cancer entities, our results suggest that the influence of miR-221 on TRAIL-mediated apoptosis is highly context- and entity-dependent.

SLC transporters as a novel class of tumour suppressors: identity, function and molecular mechanisms

Four transporters belonging to the SLC gene family (SLC5A8, SLC26A3, SLC39A1 and SLC22A18) have been identified thus far as tumour suppressors; this review summarizes the physiological functions of these transporters and the molecular mechanisms related to their tumour-suppressive function.

Επίδραση μερικής και ολικής καταστολής του διμερούς ARF-μονοπατιού απόκρισης στις δικλωνικές θραύσεις του DNA στην εξέλιξη του καρκίνου

The DNA damage response (DDR) pathway and ARF function act as barriers of human cancer development. It has been considered that the DDR and ARF exert this function independently of each other. However, a few studies propose that ARF’s activity is positively regulated by the DDR pathway. Examining this hypothesis we performed a series of experiments using molecular techniques such as immunoblotting, immunohistochemistry, immunofluorescence, immunoprecipitation, Real Time Reverse Transcritpion polymerase chain reaction (RT-PCR) in biological material from cell culture or histological samples, as well as ectopic protein expression through plasmid transfections, proteomic analyses, ribosome RNA biogenesis assay and xenografts of human cancer cells. We surprisingly found that ATM suppressed, in a transcription-independent manner, ARF protein levels and activity. Specifically, ATM activated protein phosphatase 1 (PP1). PP1 antagonized Nek2-dependent phosphorylation of nucleophosmin (NPM), liberating ARF from NPM and rendering it susceptible to degradation by the ULF E3-ubiquitin ligase. In human clinical samples, loss of ATM expression correlated with increased ARF levels and in xenograft and tissue culture models, inhibition of ATM stimulated the tumour-suppressive effects of ARF. The importance of the proposed mechanism can be exploited through a therapeutic approach, especially in cases of tumours bearing loss of p53. In such tumours, DDR may act in favour of the tumour cells, since the major effector of the antiumour barriers of apoptosis and senescence is absent, but ATM inhbition could boost ARF’s tumour-suppressive function, contributing to an anti-tumour response.