Rapid human melanoma cell death induced by sanguinarine through oxidative stress

Abstract

Sanguinarine is a natural isoquinoline alkaloid derived from the root of Sanguinaria canadensis and from other poppy fumaria species, and is known to have a broad spectrum of pharmacological properties. Here we have found that sanguinarine, at low micromolar concentrations, showed a remarkably rapid killing activity against human melanoma cells. Time-lapse videomicroscopy showed rapid morphological changes compatible with an apoptotic cell death, which was further supported by biochemical markers, including caspase activation, poly(ADP-ribose) polymerase (PARP) cleavage and DNA breakdown. Pan-caspase inhibition blocked sanguinarine-induced cell death. Sanguinarine treatment also induced an increase in intracellular calcium concentration, which was inhibited by dantrolene, and promoted cleavage of BAP-31, thus suggesting a putative role for Ca(2+) release from endoplasmic reticulum and a cross-talk between endoplasmic reticulum and mitochondria in the anti-melanoma action of sanguinarine. Sanguinarine disrupted the mitochondrial transmembrane potential (ΔΨm), released cytochrome c and Smac/DIABLO from mitochondria to cytosol, and induced oxidative stress. Overexpression of Bcl-XL by gene transfer did not prevent sanguinarine-mediated cell death, oxidative stress or release of mitochondrial apoptogenic proteins. However, preincubation with N-acetyl-l-cysteine (NAC) prevented sanguinarine-induced oxidative stress, PARP cleavage, release of apoptogenic mitochondrial proteins, and cell death. Pretreatment with glutathione (GSH) also inhibited the anti-melanoma activity of sanguinarine. Thus, pretreatment with the thiol antioxidants NAC and GSH abrogated the killing activity of sanguinarine. Taking together, our data indicate that sanguinarine is a very rapid inducer of human melanoma caspase-dependent cell death that is mediated by oxidative stress.

Antitumor effects of the benzophenanthridine alkaloid sanguinarine: Evidence and perspectives

“…Consistently, pre-treatment of tumor cells with antioxidants such as N-acetylcysteine or glutathione counteracts the apoptosis induced by sanguinarine [21,32,37,52] . Moreover, the over-expression of cyclooxygenase-2 (COX-2) also rescues prostate cancer cells from sanguinarine-induced apoptosis by sanguinarine enhances apoptosis in human mammary adenocarcinoma MCF-7 through the inhibition of VEGF release, induced by generation of ROS [32] .…”

“…Moreover, an overdone oxidative stress has been shown capable of inducing a reduction of the normal mitochondrial membrane potential, which in turn leads to apoptosis [21,[61][62][63] . It has been shown that ROS generation, is crucial for the apoptosis induced by sanguinarine in human breast cancer [52] , SK-Mel-2 human melanoma [37] , human prostate cancer [25] and in both HCT-116 [21] and HT-29…”

“…In detail, it has been reported that micromolar concentrations of sanguinarine are capable of inhibiting tumor cell growth, and this inhibitory effect is associated with cell cycle arrest and induction of apoptosis [19][20][21][22] . The anti-proliferative and/ or pro-apoptotic activities of sanguinarine have been demonstrated in in vitro studies on several cancer cell types including epidermal [23] , keratinocyte [24,25] , prostate [26][27][28] , cervical [29] , breast [20,[30][31][32][33] , leukaemia [34,35] , lymphoma [36] , melanoma [37][38][39] , colon [40,41] , colorectal [21] , gastric [42] , pancreatic [19] , lung [22] , neuroendocrine [43] , osteosarcoma [44] , and human neuroblastoma cells [45] . By contrast, there are few studies on the in vivo effectiveness of sanguinarine administration per os [46,47] in animal tumor models [33,48] .…”

Promoting Effects of Sanguinarine on Apoptotic Gene Expression in Human Neuroblastoma Cells

“…The antiproliferative and/or pro-apoptotic activities of sanguinarine have been demonstrated in cells derived from several human cancers including epidermal (Ahmad et al, 2000), keratinocytes (Adhami et al, 2003;Reagan-Shaw et al, 2006), prostate (Malikova et al, 2006b;Adhami et al, 2004;Huh et al, 2006), cervical (Ding et al, 2002), breast Debiton et al, 2003;Holy et al, 2006), leukemia (Han et al, 2008;Weerasinghe et al, 2001c;Weerasinghe et al, 2001b;Weerasinghe et al, 2001a), lymphoma (Hussain et al, 2007), melanoma (Burgeiro et al, 2013;Hammerova et al, 2011;Serafim et al, 2008), colon (Lee et al, 2012;Matkar et al, 2008), colorectal (Han et al, 2013a;Lee et al, 2012;Pica et al, 2012), gastric (Choi et al, 2009), pancreatic (Ahsan et al, 2007), lung (Jang et al, 2009), neuroendocrine (Larsson et al, 2010), osteosarcoma (Park et al, 2010), and in rat glioblastoma cells (Han et al, 2007), bladde (Han et al, 2013b). However, the impact of sanguinarine on neuroblastoma cells has not been shown.…”

Picturing Molecular Environmental Health From Mitochondria

“…For instance, extracted polysaccharides from certain edible mushrooms can cause cancer cell apoptosis in vitro by generating high level of mitochondrial ROS [60]. Other plant-derived environmental factors, such as sanguinarine and herbacetin, also are potent inducers of human cancer cell death via the ROS-mediated mitochondrial apoptotic pathway [61,62]. Finally, some environmental agents that are known to be toxic may also have beneficial effects in the aspect of inhibiting cancer cell growth.…”