Gossypol Induces Death Receptor-5 through Activation of the ROS-ERK-CHOP Pathway and Sensitizes Colon Cancer Cells to TRAIL

Abstract

Development of resistance to TRAIL, an apoptosis-inducing cytokine, is one of the major problems in its development for cancer treatment. Thus, pharmacological agents that are safe and can sensitize the tumor cells to TRAIL are urgently needed. We investigated whether gossypol, a BH3 mimetic that is currently in the clinic, can potentiate TRAIL-induced apoptosis. Intracellular esterase activity, sub-G 1 cell cycle arrest, and caspase-8, -9, and -3 activity assays revealed that gossypol potentiated TRAIL-induced apoptosis in human colon cancer cells. Gossypol also down-regulated cell survival proteins (Bclx L , Bcl-2, survivin, XIAP, and cFLIP) and dramatically up-regulated TRAIL death receptor (DR)-5 expression but had no effect on DR4 and decoy receptors. Gossypol-induced receptor induction was not cell type-specific, as DR5 induction was observed in other cell types. Deletion of DR5 by siRNA significantly reduced the apoptosis induced by TRAIL and gossypol. Gossypol induction of the death receptor required the induction of CHOP, and thus, gene silencing of CHOP abolished gossypol-induced DR5 expression and associated potentiation of apoptosis. ERK1/2 (but not p38 MAPK or JNK) activation was also required for gossypol-induced TRAIL receptor induction; gene silencing of ERK abolished both DR5 induction and potentiation of apoptosis by TRAIL. We also found that reactive oxygen species produced by gossypol treatment was critical for TRAIL receptor induction and apoptosis potentiation. Overall, our results show that gossypol enhances TRAIL-induced apoptosis through the down-regulation of cell survival proteins and the up-regulation of TRAIL death receptors through the ROS-ERK-CHOP-DR5 pathway.

Tangeretin sensitises human lung cancer cells to TRAILinduced apoptosis via ROS-JNK/ERK-CHOP pathwaymediated up-regulation of death receptor 5

“…MAPKs are important downstream mediators of ROSinduced signalling [26]. ERK, JNK, and p38 MAPK have been found to be activated in TRAILmediated apoptosis [15,30] and are involved in TRAIL receptor induction [15,26]. In our study, the significant increases in the activation of JNK and ERK, along with a considerable increase in p38 MAPK activation on exposure to tangeretin, could have been either due to direct stimulation of tangeretin and/or in part indirectly due to ROS.…”

“…The observed ROS levels could have also induced CHOP expression, because CHOP is a major stress-regulated protein [25]. Several studies have reported that chemotherapeutic agents enhance DR5 expression and TRAILmediated apoptosis via ROS-dependent pathways [15,25,26].…”

“…It has been demonstrated that the induction of DRs by various stimulants is mediated through the activation of CHOP [25,26]. We investigated whether tangeretin influenced CHOP expression and whether CHOP is also involved in DR5 expression and tangeretin-induced apoptosis.…”

“…MAPKs are important downstream mediators of ROSinduced signalling [26]. ERK, JNK, and p38 MAPK have been found to be activated in TRAILmediated apoptosis [15,30] and are involved in TRAIL receptor induction [15,26].…”

Acrolein sensitizes human renal cancer Caki cells to TRAIL-induced apoptosis via ROS-mediated up-regulation of death receptor-5 (DR5) and down-regulation of Bcl-2

“…In the previous reports, the down-regulation of c-FLIP by the peroxisome proliferator-activated receptor (PPAR)-γ agonist rosiglitazone has been shown to sensitize cells to TRAIL [34]. Gossypol, a polyphenol derived from cottonseed oil, down-regulated the expression of Bcl-2, Bcl-xL and survivin, all of which have been linked to tumor cells to TRAIL [35]. Acrolein also down-regulated the expression of Bcl-2 (Fig.…”