The Effect of Bornyl cis-4-Hydroxycinnamate on Melanoma Cell Apoptosis Is Associated with Mitochondrial Dysfunction and Endoplasmic Reticulum Stress

Abstract

Bornyl cis-4-hydroxycinnamate, an active compound isolated from Piper betle stems, was investigated in terms of its effects on A2058 and A375 melanoma cell proliferation and protein expression in this study. We used flow cytometric analysis to examine the early stages of apoptosis induced by bornyl cis-4-hydroxycinnamate in the two melanoma cell lines and employed comparative proteomic analysis to investigate the effects of this compound on protein expression in A375 cells. Master maps generated by PDQuest software from two-dimensional electrophoresis (2-DE) analysis of A375 cells showed that the expression levels of 35 proteins were significantly altered, with 18 proteins upregulated and 17 downregulated. The proteomics study identified several proteins that are involved in mitochondrial dysfunction and endoplasmic reticulum stress (ER stress), in addition to apoptosis-associated proteins, including prohibitin, hypoxia-upregulated protein 1, stress 70 protein, 78 kDa glucose-regulated protein (GRP78), and protein deglycase DJ-1 (protein DJ-1) in melanoma cells exposed to bornyl cis-4-hydroxycinnamate. The treatment also resulted in a marked decline of the mitochondrial membrane potential, in cytochrome C release into the cytosol, in the activation of Bcl-2-associated X protein (Bax), Bcl-2-associated death promoter protein (Bad), caspase-3, and caspase-9, and in the decreased expression of p-Bad, B-cell lymphoma 2 (Bcl-2), Bcl-xl, and induced myeloid leukemia cell differentiation protein-1 (Mcl-1), indicating that apoptosis induced by bornyl cis-4-hydroxycinnamate was mediated by the mitochondria through the caspase-dependent pathway. Also, salubrinal (an eukaryotic initiation factor 2α inhibitor; eIF2α inhibitor) was able to protect the cells from bornyl cis-4-hydroxycinnamate-induced apoptosis. Bornyl cis-4-hydroxycinnamate-related cell death also implied that the protein kinase R-like endoplasmic reticulum kinase (PERK)–eIF2α–ATF4–CHOP signal pathways was activated upon bornyl cis-4-hydroxycinnamate treatment. Altogether, our results support the conclusion that bornyl cis-4-hydroxycinnamate-induced apoptosis in melanoma cells is associated with mechanisms correlated with the activation of caspase cascades, mitochondrial dysfunction, and endoplasmic reticulum stress, and indicate that this molecule has the potential to be developed as a chemotherapeutic agent for human melanoma.

Erythropoietin Derived Peptide Improved Endoplasmic Reticulum Stress and Ischemia-Reperfusion Related Cellular and Renal Injury

“…Therefore, our data demonstrated that CHOP involved in renal IR-related and IR injuries, and CHBP-induced renoprotection might be through regulating CHOP or improving ERS. However, when UPR response cannot restore the homeostasis of endoplasmic reticulum upon IR injury, PERK pathway could trigger off downstream apoptosis via the activation of CHOP, caspase-12 and Bcl-2 family (44,47,48). Additional correlation analysis revealed that CHOP protein was positively correlated with the level of TID and active caspase-3 protein at 2 weeks.…”

Bisphenol A Exposure Changes the Transcriptomic and Proteomic Dynamics of Human Retinoblastoma Y79 Cells

“…This protein localizes to the endoplasmic reticulum and mitochondria, and it protects cells from cell stress responses, including oxidative stress and unfolded protein responses [ 52 , 53 ]. Yang et al has recently demonstrated that mitochondria dysfunction and endoplasmic reticulum stress bornyl cis-4-hydroxycinnamate exposure occurs upregulated HYOU1 expression in melanoma cells leading to induction of apoptosis [ 54 ]. Furthermore, it has been demonstrated that HYOU1 is related to various pathologic situations, such as ischemic brain [ 55 ], malignant tumors [ 56 ].…”

Cannabidiol Affects Extracellular Vesicle Release, miR21 and miR126, and Reduces Prohibitin Protein in Glioblastoma Multiforme Cells

“…Interestingly, PHB accumulation occurs in mitochondria after chemotherapy treatment and de-novo accumulation has been shown to be associated with chemoresistance in melanoma in vitro, while knock-down of PHB sensitized melanoma cells to chemotherapy [64]. Changes in PHB levels have also been associated with melanoma cell proliferation, mitochondrial dysfunction, ER stress and melanoma cell apoptosis, in response to bornyl cis-4-Hydroxycinnamate from Piper betle stems [81].…”