Harmine activates intrinsic and extrinsic pathways of apoptosis in B16F-10 melanoma

Abstract

BackgroundHarmine is a beta-carboline alkaloid from the plant Peganum harmala. Previous studies found that harmine inhibited metastasis of B16F-10 melanoma cells. This study aims to elucidate the role of harmine in apoptosis of B16F-10 cells.MethodsB16F-10 melanoma cells were treated in the presence and absence of harmine in vitro. Morphological changes, cell cycle and expression of various pro and anti- apoptotic genes were analyzed for the study of apoptosis.ResultsMorphological observation and DNA laddering assay showed that harmine treated cells displayed marked apoptotic characteristics, such as nuclear fragmentation, appearance of apoptotic bodies and DNA laddering fragment. TUNEL assay and flow cytometric analysis also confirmed apoptosis. Furthermore, RT-PCR analysis showed that harmine induced apoptosis in B16F-10 melanoma cells by up-regulating Bax and activating Caspase-3, 9 and p53 and down-regulating Bcl-2. Harmine also up-regulated Caspase-8 and Bid, indicating that harmine affected both extrinsic and intrinsic pathways of apoptosis. This study also showed inhibitory effects of harmine on some transcription factors and pro- inflammatory cytokines that protect cell from apoptosis.ConclusionHarmine activates both intrinsic and extrinsic pathways of apoptosis and regulates some transcription factors and pro-inflammatory cytokines.

Amentoflavone Induces Apoptosis and Inhibits NF-ĸBmodulated Anti-apoptotic Signaling in Glioblastoma Cells

“…Protein expression of MCL1 and C-FLIP are modulated by NF-ĸB activation in cancer (12). Han et al showed that silencing NF-ĸB reduced C-FLIP expression and enhanced TRAIL-induced apoptosis in glioblastoma cells (24 TRAIL in glioblastoma cells (6). We confirmed such effects by finding that NF-ĸB inhibitor reduced NF-ĸB activation and protein expression of MCL1 and C-FLIP in U-87 MG cells (Figure 3).…”

A review on medicinal importance, pharmacological activity and bioanalytical aspects of beta-carboline alkaloid “Harmine”

“…Harmine has been reported to have antidepressantlike actions in rodents [2] . Harmine possesses anxiolytic, behavioral effects and anti-tumor potential both in vitro and in vivo [5] . Harmine has high inhibitory affinity of DYRK1A kinase activity, suggesting that harmine could alter tau phosphorylation [6] .…”

“…The role of harmine in apoptosis of B16F-10 cells was investigated. Harmine activates both intrinsic and extrinsic pathways of apoptosis and regulates some transcription factors and pro-inflammatory cytokines [5] . The in vivo anti-angiogenic activity of harmine was studied using B16F-10 melanoma cells in C57BL/6 mice.…”

“…Harmine is widely distributed in nature, such as in various plants, marine creatures, insects, mammalians, human tissues and body fluids. Harmine have antimicrobial, antiplasmodial, antifungal, antioxidative, antitumor, antimutagenic, cytotoxic and hallucinogenic properties [1][2][3][4][5][6][7] . Beta-carboline compounds act as inverse agonists at the benzodiazepine site of the gammaaminobutyric acid type A receptors and have actions entirely opposite to those of the anxiolytic benzodiazepines.…”

Antitumor properties and toxicity effects of Peganum harmala L. (Zygophyllaceae)

“…These in vitro studies have shown that harmine, b-carbolines alkaloid have more activity (Lamchouri et al, 2010). The harmine and β-carbolines alkaloid are highly cytotoxic and significantly inhibited tumor cell growth with apoptotic effect (Hamsa & Kuttan, 2011). This study showed that harmine significantly inhibited tumor nodule formation in the lung tissue and decreased various biochemical parameters associated with lung metastasis.…”