Induction of apoptotic cell death in human bladder cancer cells by ethanol extract ofZanthoxylum schinifoliumleaf, through ROS-dependent inactivation of the PI3K/Akt signaling pathway

Abstract

BACKGROUND/OBJECTIVES Zanthoxylum schinifolium is traditionally used as a spice for cooking in East Asian countries. This study was undertaken to evaluate the anti-proliferative potential of ethanol extracts of Z. schinifolium leaves (EEZS) against human bladder cancer T24 cells. MATERIALS/METHODS Subsequent to measuring the cytotoxicity of EEZS, the anti-cancer activity was measured by assessing apoptosis induction, reactive oxygen species (ROS) generation, and mitochondrial membrane potential (MMP). In addition, we determined the underlying mechanism of EEZS-induced apoptosis through various assays, including Western blot analysis. RESULTS EEZS treatment concentration-dependently inhibited T24 cell survival, which is associated with apoptosis induction. Exposure to EEZS induced the expression of Fas and Fas-ligand, activated caspases, and subsequently resulted to cleavage of poly (ADP-ribose) polymerase. EEZS also enhanced the expression of cytochrome c in the cytoplasm by suppressing MMP, following increase in the ratio of Bax:Bcl-2 expression and truncation of Bid. However, EEZS-mediated growth inhibition and apoptosis were significantly diminished by a pan-caspase inhibitor. Moreover, EEZS inhibited activation of the phosphoinositide 3-kinase (PI3K)/Akt pathway, and the apoptosis-inducing potential of EEZS was promoted in the presence of PI3K/Akt inhibitor. In addition, EEZS enhanced the production of ROS, whereas N-acetyl cysteine (NAC), a ROS scavenger, markedly suppressed growth inhibition and inactivation of the PI3K/Akt signaling pathway induced by EEZS. Furthermore, NAC significantly attenuated the EEZS-induced apoptosis and reduction of cell viability. CONCLUSIONS Taken together, our results indicate that exposure to EEZS exhibits anti-cancer activity in T24 bladder cancer cells through ROS-dependent induction of apoptosis and inactivation of the PI3K/Akt signaling pathway.

A Cross-Talk about Radioresistance in Lung Cancer—How to Improve Radiosensitivity According to Chinese Medicine and Medicaments That Commonly Occur in Pharmacies

“…Moreover, extracts increase the expression of cytochrome c by MMP suppression related to an increased Bax:Bcl-2 ratio. Finally, Zanthoxylum extract potentiates ROS production and inhibits the PI3K/Akt signaling pathway [ 47 ] ( Figure 1 ). However, less is known about schinifoline’s pharmacological activities separately.…”

“…: decreases levels of EGFR, ErB2 and surviving Apoptosis induction: up-regulation of P53 protein, potentiated cytochrome c release from mitochondria, cleaved caspases for instance caspase-3, caspase-8, caspase-9 and PARP[31,32] Tashinone Apoptosis induction: p53, Bax p-21, suppressed production of anti-apoptotic proteins such as Bcl-2, c-Myc Autophagy induction: c-Jun N-terminal kinase activation With IR: PPAT downregulation[34][35][36][37] DiosmetinApoptosis induction: increasing of ROS level, inhibition of Nrf2 by disruption stability through Keap1-mediated proteasomal degradation, G1/S phase arrest of the cycle With IR: IR-induced DNA damage repair inhibition, inhibition of Akt signaling pathway, induction of G1 phase arrest[38][39][40][41][42] SchinifolineApoptosis induction: Production of Fas and FasL, pro-apoptotic caspases, cytochrome c, increased bax:Bcl-2 ratio, stimuli ROS production, inhibition of PI3K/Akt signaling pathway, G2/M arrest[43,44] Huachansu Apoptosis induction: enhance of ROS level, activation of the G 2 /M checkpoint, extending attendance of γH2AX foci, up-regulation cleaved of caspase-3 and cleaved poly-(ADP-ribose) polymerase (PARP), inhibition of Bcl-2 and P53 protein, suppression of TopoII[45,46] Metformin Apoptosis induction: inhibition of mTOR, suppression of c-Myc and Hif-1α, AMPK activation with IR: maintain activation of ATM-AMPK-p53/p21 cip1 and inhibition of Akt-mTOR-4EBP1, through KEAP1-independent mechanism, causes decrease of NRF2 due to ubiquitination and proteasomal degradation, preferentially acts with CSC Other with IR: protect from fibrosis and pneumonitis[47][48][49][50][51][52][53][54] Lovastatin Apoptosis induction: lower-level of glutathione, increase of cytochrome c, caspase-3, p53, bax, COX-2 activation and PPARγ with IR: Akt inhibition, stimulation of Erk phosphorylation, block expression p53 protein and kinase inhibitors p21 cip1 and p27 kip Other with IR: in normal epithelial tissue acts radioprotective, interact with Rho small GTPases protect lung epithelial, fibroblasts, microvascular epithelial cells against IR-induce toxicity by improved repair DNA mechanisms and caspase-dependent apoptosis[55][56][57]…”