For example, increased hepatocyte growth factor signaling through c-MET, increased MK0683 mouse susceptibility to TGF-α/EGF signaling, as well as modifications in extracellular matrix turnover and remodeling are implicated in the pathogenesis of RCC [40]. Clearly, RCC is a MX69 mw complex disease resulting from numerous alterations of genes and pathways that work in concert, indicating that pursuing a single target or pathway will not yield chemotherapeutics with significant efficacy. The best chance for achieving therapeutic efficacy in a disease

such as RCC should involve the use of agents that target the multiple pathways which contribute fundamentally to this disease. Natural products are well known to affect multiple targets and thus have excellent potential as chemotherapeutic agents. The relatively recently identified natural product, englerin (EA), is very unique due to its high selectivity against RCC that is 1000-fold higher than any other cell type [16]. Our results demonstrate that EA induces apoptosis and autophagy in addition to necrosis in A498 RCC cells at nanomolar concentrations. This finding is in contrast to a recent report stating that EA induced necrosis but

not apoptosis or autophagy [22]. 4SC-202 mouse In this previous study, however, autophagy was most likely inhibited by the supplementation of culture medium with non-essential amino acids (NEAA), a known inhibitor of autophagy [41], and was thus not observed. Our results confirmed that autophagy induced by EA

could be inhibited by NEAA. We further showed that inhibition of autophagy by NEAA did not diminish cell death. This finding is supported by the previous study which showed that RCC cells died under conditions which inhibited autophagy with a sensitivity to EA similar to that observed by us and others [16, 21]. For instance, in viability assays in the study by Sulzmaier et al. [22], EA was found to have an EC50 of 53 nM in the presence of NEAA. In the absence of NEAA, the estimated EC50 of EA in A498 cells in our viability assay was 63 nM (Figure 1 and data Inositol monophosphatase 1 not shown). Furthermore, the NCI reported LC50 for EA in A498 cells, under conditions not inhibiting autophagy, was 79 nM [16]. Though the NCI determined LC50 is a somewhat different measure than the EC50, determined by us and Sulzmaier et al. [22], in addition to the assays being different, the fact that these values are not very different regardless of whether autophagy is inhibited, indicates that autophagy does not appear to have much of an effect on cell death. Though autophagy can play a pro-death role when prolonged or in certain developmental conditions [42], in most circumstances, autophagic generation of nutrients prevents or delays cell death [43], thus acting as a survival mechanism.