Effects of statins and farnesyl transferase inhibitors on <scp>ERK</scp> phosphorylation, apoptosis and cell viability in non‐small lung cancer cells

Abstract

Abstract Objective 3‐hydroxy‐3‐methylglutaryl‐coenzyme A ( HMG ‐CoA) reductase inhibitors (statins) can affect post‐translational processes, thus being responsible for decreased farnesylation and geranylgeranylation of intracellular small G proteins such as Ras, Rho and Rac, essential for cell survival and proliferation. In this regard, recent in vitro and in vivo studies suggest a possible role for both statins and farnesyl transferase inhibitors in the treatment of malignancies. Within such a context, the aim of our study was to investigate effects of either simvastatin (at concentrations of 1, 15, and 30 μ m ) or the farnesyl transferase inhibitor R 115777 (at concentrations of 0.1, 1, and 10 μ m ), on two cultures of human non‐small lung cancer cells, adenocarcinoma ( GLC ‐82) and squamous ( CALU ‐1) cell lines. In particular, we evaluated actions of these two drugs on phosphorylation of the ERK 1/2 group of mitogen‐activated protein kinases and on apoptosis, plus on cell numbers and morphology. Materials and Methods Western blotting was used to detect ERK phosphorylation, and to assess apoptosis by evaluating caspase‐3 activation; apoptosis was also further assessed by terminal deoxynucleotidyl‐mediated d UTP nick end labelling ( TUNEL ) assay. Cell counting was performed after trypan blue staining. Results and conclusion In both GLC ‐82 and CALU ‐1 cell lines, simvastatin and R 115777 significantly reduced ERK phosphorylation; this effect, which reached the greatest intensity after 36 h treatment, was paralleled by a concomitant induction of apoptosis, documented by significant increase in both caspase‐3 activation and TUNEL ‐positive cells, associated with a reduction in cell numbers. Our results thus suggest that simvastatin and R 115777 may exert, in susceptible lung cancer cell phenotypes, a pro‐apoptotic and anti‐proliferative activity, which appears to be mediated by inhibition of the Ras/Raf/ MEK / ERK signalling cascade.