c‐Jun N‐terminal Kinase‐Dependent Endoplasmic Reticulum Stress Pathway is Critically Involved in Arjunic Acid Induced Apoptosis in Non‐Small Cell Lung Cancer Cells

Though arjunic acid, a triterpene isolated from Terminalia arjuna, was known to have antioxidant, antiinflammatory, and cytotoxic effects, its underlying antitumor mechanism still remains unclear so far. Thus, in the present study, the molecular antitumor mechanism of arjunic acid was examined in A549 and H460 non‐small cell lung cancer (NSCLC) cells. Arjunic acid exerted cytotoxicity by 3‐[4, 5‐dimethylthiazol‐2‐yl]‐2, 5‐diphenyl tetrazolium bromide (MTT) assay and significantly increased sub‐G1 population in A549 and H460 cells by cell cycle analysis. Consistently, arjunic acid cleaved poly (ADP‐ribose) polymerase (PARP), activated Bax, and phosphorylation of c‐Jun N‐terminal kinases (JNK), and also attenuated the expression of pro‐caspase‐3 and Bcl‐2 in A549 and H460 cells. Furthermore, arjunic acid upregulated the expression of endoplasmic reticulum (ER) stress proteins such as IRE1 α, ATF4, p‐eIF2α, and C/EBP homologous protein (CHOP) in A549 and H460 cells. Conversely, CHOP depletion attenuated the increase of sub‐G1 population by arjunic acid, and also JNK inhibitor SP600125 blocked the cytotoxicity and upregulation of IRE1 α and CHOP induced by arjunic acid in A549 and H460 cells. Overall, our findings suggest that arjunic acid induces apoptosis in NSCLC cells via JNK mediated ER stress pathway as a potent chemotherapeutic agent for NSCLC.     

Auraptene Induces Apoptosis via Myeloid Cell Leukemia 1‐Mediated Activation of Caspases in PC3 and DU145 Prostate Cancer Cells

Although auraptene, a prenyloxy coumarin from Citrus species, was known to have anti‐oxidant, anti‐bacterial, antiinflammatory, and anti‐tumor activities, the underlying anti‐tumor mechanism of auraptene in prostate cancers is not fully understood to date. Thus, in the present study, we have investigated the anti‐tumor mechanism of auraptene mainly in PC3 and DU145 prostate cancer cells, because auraptene suppressed the viability of androgen‐independent PC3 and DU145 prostate cancer cells better than androgen‐sensitive LNCaP cells. Also, auraptene notably increased sub‐G1 cell population and terminal deoxynucleotidyl transferase dUTP nick end labeling‐positive cells as features of apoptosis in two prostate cancer cells compared with untreated control. Consistently, auraptene cleaved poly(ADP‐ribose) polymerase, activated caspase‐9 and caspase‐3, suppressed the expression of anti‐apoptotic proteins, including Bcl‐2 and myeloid cell leukemia 1 (Mcl‐1), and also activated pro‐apoptotic protein Bax in both prostate cancer cells. However, Mcl‐1 overexpression reversed the apoptotic effect of auraptene to increase sub‐G1 population and induce caspase‐9/3 in both prostate cancer cells. Taken together, the results support scientific evidences that auraptene induces apoptosis in PC3 and DU145 prostate cancer cells via Mcl‐1‐mediated activation of caspases as a potent chemopreventive agent for prostate cancer prevention and treatment. Copyright © 2017 John Wiley & Sons, Ltd.     

Obovatol Induces Apoptosis in Non‐small Cell Lung Cancer Cells via C/EBP Homologous Protein Activation

Although obovatol, a phenolic compound from the bark of Magnolia obovata, was known to have antioxidant, neuroprotective, antiinflammatory, antithrombotic and antitumour effects, its underlying antitumour mechanism is poorly understood so far. Thus, in the present study, the antitumour molecular mechanism of obovatol was investigated in non‐small cell lung cancer cells (NSCLCs). Obovatol exerted cytotoxicity in A549 and H460 NSCLCs, but not in BEAS‐2B cells. Also, obovatol increased sub‐G1 accumulation and early and late apoptotic portion in A549 and H460 NSCLCs. Consistently, obovatol cleaved PARP, activated caspase 9/3 and Bax and attenuated the expression of cyclin D1 in A549 and H460 NSCLCs. Interestingly, obovatol upregulated the expression of endoplasmic reticulum stress proteins such as C/EBP homologous protein (CHOP), IRE1α, ATF4 and p‐elF2 in A549 and H460 NSCLCs. Conversely, depletion of CHOP blocked the apoptotic activity of obovatol to increase sub‐G1 accumulation in A549 and H460 NSCLCs. Overall, our findings support scientific evidences that obovatol induces apoptosis via CHOP activation in A549 and H460 NSCLCs. Copyright © 2016 John Wiley & Sons, Ltd.     

Activation of Caspase‐9/3 and Inhibition of Epithelial Mesenchymal Transition are Critically Involved in Antitumor Effect of Phytol in Hepatocellular Carcinoma Cells

This study was designed to investigate the antitumor mechanism of Phytol in hepatocellular carcinomas including Huh7 and HepG2 cells in association with caspase dependent apoptosis and epithelial mesenchymal transition (EMT) signaling. Phytol significantly suppressed the viability of Huh7 and HepG2 cells. Also, Phytol significantly increased the sub G1 population and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) positive cells in a concentration dependent manner in Huh7 and HepG2 cells. Consistently, Phytol cleaved poly (adenosine diphosphate‐ribose) polymerase (PARP), activated caspase‐9/3, and Bax attenuated the expression of survival genes such as Bcl‐2, Mcl‐1, and c‐Myc in Huh7 and HepG2 cells. Of note, Phytol also suppressed typical morphology change of EMT such as loss of cell adhesion and formation of fibroblast like mesenchymal cells in HepG2 cells. Furthermore, Phytol also reversed the loss of E‐cadherin and overexpression of p‐smad2/3, alpha‐smooth muscle actin, and Snail induced by EMT promoter transforming growth factor beta1 in HepG2 cells. Overall, our findings suggest that Phytol exerts antitumor activity via apoptosis induction through activation of caspas‐9/3 and inhibition of EMT in hepatocellular carcinoma cells as a potent anticancer candidate for liver cancer treatment. Copyright © 2015 John Wiley & Sons, Ltd.     

Ocimum sanctum induces apoptosis in A549 lung cancer cells and suppresses the in vivo growth of lewis lung carcinoma cells

Although Ocimum sanctum has been used extensively for its medicinal values in India and China, its antitumor activity against human nonsmall cell lung carcinoma (NSCLC) A549 cells has not been investigated until now. Therefore, the antitumor mechanism of ethanol extracts of Ocimum sanctum (EEOS) was elucidated in A549 cells in vitro and the Lewis lung carcinoma (LLC) animal model. EEOS exerted cytotoxicity against A549 cells, increased the sub‐G1 population and exhibited apoptotic bodies in A549 cells. Furthermore, EEOS cleaved poly(ADP‐ribose)polymerase (PARP), released cytochrome C into cytosol and simultaneously activated caspase‐9 and ‐3 proteins. Also, EEOS increased the ratio of proapoptotic protein Bax/antiapoptotic protein Bcl‐2 and inhibited the phosphorylation of Akt and extracellular signal regulated kinase (ERK) in A549 cancer cells. In addition, it was found that EEOS can suppress the growth of LLC inoculated onto C57BL/6 mice in a dose‐dependent manner. Overall, these results demonstrate that EEOS induces apoptosis in A549 cells via a mitochondria caspase dependent pathway and inhibits the in vivo growth of LLC, suggesting that EEOS can be applied to lung carcinoma as a chemopreventive candidate. Copyright © 2009 John Wiley & Sons, Ltd.     

Inhibition of Myeloid Cell Leukemia 1 and Activation of Caspases Are Critically Involved in Gallotannin‐induced Apoptosis in Prostate Cancer Cells

Although gallotannin contained in several medicinal plants was known to have multi‐biological activities, such as antioxidant, antiinflammatory, antimicrobial, immunomodulatory, and antitumor effects, the underlying apoptotic mechanism of gallotannin is not fully understood so far. Thus, in the present study, the apoptotic mechanism of gallotannin was elucidated in DU145, PC‐3, and M2182 prostate cancer cells in association with myeloid cell leukemia 1 (Mcl‐1) signaling. Gallotannin exerted dose‐dependent cytotoxicity in DU145, PC‐3, and M2182 prostate cancer cells. Also, gallotannin showed apoptotic morphological features and increased the number of terminal deoxynucleotidyl transferase dUTP nick end labeling positive cells and sub‐G1 accumulation in three prostate cancer cell lines. Consistently, gallotannin cleaved poly (ADP‐ribose) polymerase (PARP) and attenuated the expression of procaspases 9 and 3 in three prostate cancer cell lines. Furthermore, gallotannin attenuated the expression of survival genes such as Mcl‐1, B‐cell lymphoma 2, and B‐cell lymphoma 2 extra large in three prostate cancer cell lines. Interestingly, overexpression of Mcl‐1 reversed the ability of gallotannin to cleave PARP and increase sub‐G1 population in three prostate cancer cell lines. Conversely, silencing of Mcl‐1 enhanced apoptosis by gallotannin in three prostate cancer cell lines by FACSCalibur (Becton Dickinson, Franklin Lakes, NJ, USA). Taken together, our findings demonstrate that inhibition of Mcl‐1 and activation of caspases are critically involved in gallotannin‐induced apoptosis in prostate cancer cells. Copyright © 2015 John Wiley & Sons, Ltd.     

Effect of Diphlorethohydroxycarmalol Isolated From Ishige okamurae on Apoptosis in 3 t3‐L1 Preadipocytes

Obesity is an important issue in the world of public health and preventive medicine. Inhibition of proliferation of preadipocytes plays an important role in proposed antiobesity mechanisms. In this study, we investigated the effect of diphlorethohydroxycarmalol (DPHC) isolated from Ishige okamurae on the apoptotic pathway. The results showed that DPHC inhibited population growth in 3 T3‐L1 preadipocytes as assessed using the 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay. Flow cytometric analysis of 3 T3‐L1 preadipocytes showed that the number of early and late apoptotic cells increases in a dose‐dependent manner after exposure to DPHC, while the number of normal cells was reduced. Our findings indicate that the induction of apoptosis in 3 T3‐L1 preadipocytes by DPHC is mediated through the activation of caspase‐3, Bax, and caspase‐9, and then through the cleavage of poly(ADP‐ribose) polymerase and the down‐regulation of Bcl‐2. The data also indicated that treatment with DPHC inhibits histone deacetylase activity in 3 T3‐L1 preadipocytes. These results show that DPHC efficiently induces apoptosis in 3 T3‐L1 preadipocytes. Copyright © 2012 John Wiley & Sons, Ltd.     

Apoptotic Effect of Astragalin in Melanoma Skin Cancers via Activation of Caspases and Inhibition of Sry‐related HMg‐Box Gene 10

Though Astragalin (kaempferol‐3‐glucoside) contained in Paeonia lactiflora and other plants was known to have anti‐oxidant, antiinflammatory, and anti‐tumor activity, the anti‐tumor mechanism of Astragalin has never been reported in melanomas until now. Thus, in the present study, the underlying apoptotic mechanism of Astragalin isolated from Aceriphyllum rossii was elucidated in A375P and SK‐MEL‐2 melanoma cells. Astragalin exerted cytotoxicity in A375P and SK‐MEL‐2 cells in a concentration‐dependent manner. Also, Astragalin significantly increased the number of TdT‐mediated dUTP nick end labeling positive cells and sub‐G1 population as a feature of apoptosis in A375P and SK‐MEL‐2 cells compared with untreated control. Consistently, western blotting revealed that Astragalin activated caspase 9/3 and Bax, cleaved poly (ADP‐ribose) polymerase, and attenuated the expression of cyclin D1, Mcl‐1, and Sry‐related HMg‐Box gene 10 (SOX10) in A375P and SK‐MEL‐2 cells. Of note, ectopic expression of SOX10 reduced the apoptotic ability of Astragalin to inhibit proliferation, cleave poly (ADP‐ribose) polymerase, and caspase 3 in A375P and SK‐MEL‐2 melanoma cells. Overall, our findings provide evidence that Astragalin induces apoptosis in A375P and SK‐MEL‐2 melanoma cells via activation of caspase9/3 and inhibition of SOX10 signaling. Copyright © 2017 John Wiley & Sons, Ltd.     

Apoptotic Effect of Sanggenol L via Caspase Activation and Inhibition of NF‐κB Signaling in Ovarian Cancer Cells

In the present study, the underlying apoptotic mechanism of sanggenol L was elucidated in ovarian cancer cells. Sanggenol L showed cytotoxic and antiproliferative effect in A2780, SKOV‐3, and OVCAR‐3 ovarian cancer cells in a concentration‐dependent fashion. Consistently, sanggenol L increased sub‐G1 phase population and early and late apoptotic portion in ovarian cancer cells. Also, sanggenol L activated caspase9/3, suppressed the phosphorylation of IκBα and p65 NF‐κB (nuclear factor kappa‐light‐chain‐enhancer of activated B cells), attenuated the expression of Cyclin D1, and cleaved poly(adenosine diphosphate ribose ‐ribose) polymerase in SKOV‐3, A2780, and OVCAR‐3 cells. Furthermore, sanggenol L blocked nuclear translocation of NF‐κB and also attenuated the expression of NF‐κB related genes such as c‐Myc, Cyclin D1, and Bcl‐_XL, Bcl‐2, in lipopolysaccharide‐treated SKOV‐3 cells. Overall, our findings for the first time suggest that sanggenol L induces apoptosis via caspase activation and inhibition of NF‐κB/IκBα phosphorylation as a potent chemotherapeutic agent for ovarian cancers. Copyright © 2015 John Wiley & Sons, Ltd.     

Phloroglucinol Protects INS‐1 Pancreatic β‐cells Against Glucotoxicity‐Induced Apoptosis

Decreasing numbers, and impaired function, of pancreatic β‐cells are key factors in the development of type 2 diabetes. This study was designed to investigate whether phloroglucinol protected pancreatic β‐cells against glucotoxicity‐induced apoptosis using a rat insulinoma cell line (INS‐1). High glucose treatment (30 mM) induced INS‐1 cell death; however, the level of glucose‐induced apoptosis was significantly reduced in cells treated with 100‐μM phloroglucinol. Treatment with 10–100‐μM phloroglucinol increased cell viability and decreased intracellular levels of reactive oxygen species, nitric oxide, and lipid peroxidation dose‐dependently in INS‐1 cells pretreated with high glucose. Furthermore, phloroglucinol treatment markedly reduced the protein expression of Bax, cytochrome c, and caspase 9, while increasing anti‐apoptotic Bcl‐2 protein expression. Cell death type was examined using annexin V/propidium iodide staining, revealing that phloroglucinol markedly reduced high glucose‐induced apoptosis. These results demonstrated that phloroglucinol could be useful as a potential therapeutic agent for the protection of pancreatic β‐cells against glucose‐induced apoptosis. Copyright © 2015 John Wiley & Sons, Ltd.     

Pachymic Acid Induces Apoptosis of EJ Bladder Cancer Cells by DR5 Up‐Regulation,ROS Generation,Modulation of Bcl‐2 and IAP Family Members

Pachymic acid (PA) is a lanostane‐type triterpenoid derived from Poria cocos mushroom that possess various biological effects such as anti‐cancer, antiinflammatory and anti‐metastasis effects. In this study, we investigated the anti‐cancer effects of PA in EJ bladder cancer cells. The results showed that PA significantly inhibited proliferation of EJ cells in a dose‐dependent manner. PA induced accumulation of sub‐G1 DNA content (apoptotic cell population), apoptotic bodies and chromatin condensation and DNA fragmentation in EJ cells in a dose‐dependent manner. PA also induces activation of caspase‐3, ‐8 and ‐9, and subsequent cleavage of poly (ADP‐ribose) polymerase, and significantly suppressed the inhibitor of apoptosis protein family proteins in a dose‐dependent manner. Furthermore, PA activates Bid and induced the loss of mitochondrial membrane potential (ΔΨ_m) with up‐regulated pro‐apoptotic proteins (Bax and Bad), down‐regulated anti‐apoptotic proteins (Bcl‐2 and Bcl‐xL) and cytochrome c release. In turn, PA increased the generation of reactive oxygen species (ROS); also, the ROS production was blocked by N‐acetyl‐L‐cysteine. The expressions of TNF‐related apoptosis inducing ligand and death receptor 5 were up‐regulated by PA in a dose‐dependent manner, suggesting extrinsic pathway also involved in PA‐induced apoptosis. This study provides evidence that PA might be useful in the treatment of human bladder cancer. Copyright © 2015 John Wiley & Sons, Ltd.     

7,3′‐dimethoxy hesperetin induces apoptosis of fibroblast‐like synoviocytes in rats with adjuvant arthritis through caspase 3 activation

Approaches inducing fibroblast‐like synoviocytes (FLS) apoptosis in rheumatoid arthritis (RA) patients have been considered as a promising strategy for treating RA. Here, adjuvant arthritis (AA) in rat was induced by complete Freund's adjuvant and FLS were separated and cultured using a tissue explant cultivation method. The apoptotic effect of 7,3′‐dimethoxy hesperetin (DMHP, a highly antirheumatic active derivative of hesperidin) on AA FLS was evaluated with MTT assay, Hoechst staining and flow cytometry analysis. Bcl‐2, Bax, caspase 3 gene expressions and caspase 3 activity were assayed to identify whether caspase 3 was involved in the apoptosis induced by DMHP. It was found that DMHP significantly decreased AA FLS proliferation in vitro by MTT assay. The AA FLS treated with DMHP displayed typical apoptotic characteristics including irregularity in shape, nuclear shrinkage and chromatin condensation. Flow cytometry analysis indicated that DMHP could obviously increase the AA FLS apoptosis rate. Compared with the AA‐FLS control group, DMHP markedly decreased the mRNA expression of Bcl‐2, whereas those of Bax and caspase 3 were increased. Moreover, DMHP significantly increased caspase 3 activity in a dose‐dependent manner. In aggregate, the results demonstrate that DMHP effectively induces AA FLS apoptosis through caspase 3 activation and can be considered as a possible antirheumatic agent. Copyright © 2010 John Wiley & Sons, Ltd.     

Liquiritigenin induces mitochondria‐mediated apoptosis via cytochrome c release and caspases activation in heLa Cells

It has been demonstrated that many flavonoids possess a potent and broad spectrum of antitumor activity. Liquiritigenin is a flavanone extracted from Glycyrrhizae. This study investigated the effects of liquiritigenin on cell viability and apoptosis induction in human cervical carcinoma (HeLa) cells. The results show that liquiritigenin significantly suppressed cell proliferation in a dose‐ and time‐dependent manner in HeLa cells. In addition, liquiritigenin promoted apoptosis in HeLa cells, evidenced by apoptotic morphological changes and Annexin‐V binding. The apoptosis induction with liquiritigenin is associated with the up‐regulation of p53 and Bax, along with down‐regulation of Bcl‐2 and survivin. Finally, examination of the mitochondrial pathway of apoptosis revealed that cytochrome c is released from mitochondria to cytosol, associated with the activation of caspase‐9 and ‐3, and the cleavage of poly (ADP‐ribose) polymerase (PARP). Overall, the results indicate that liquiritigenin induces apoptosis in part via the mitochondrial pathway, which is associated with p53 up‐regulation, release of cytochrome c and elevated activity of caspase‐9 and ‐3 in HeLa cells. Copyright © 2010 John Wiley & Sons, Ltd.     

Morin enhances auranofin anticancer activity by up‐regulation of DR4 and DR5 and modulation of Bcl‐2 through reactive oxygen species generation in Hep3B human hepatocellular carcinoma cells

Evidence suggests that auranofin (AF) exhibits anticancer activity by inhibiting thioredoxin reductase (TrxR). Here, in this study, we have investigated the synergistic effects of AF and morin and their mechanism for the anticancer effects focusing on apoptosis in Hep3B human hepatocellular carcinoma cells. We assessed the anticancer activities by annexin V/PI double staining, caspase, and TrxR activity assay. Morin enhances the inhibitory effects on TrxR activity of AF as well as reducing cell viability. Annexin V/PI double staining revealed that morin/AF cotreatment induced apoptotic cell death. Morin enhances AF‐induced mitochondrial membrane potential (ΔΨm) loss and cytochrome c release. Further, morin/AF cotreatment upregulated death receptor DR4/DR5, modulated Bcl‐2 family members (upregulation of Bax and downregulation of Bcl‐2), and activated caspase‐3, ‐8, and ‐9. Morin also enhances AF‐induced reactive oxygen species (ROS) generation. The anticancer effects results from caspase‐dependent apoptosis, which was triggered via extrinsic pathway by upregulating TRAIL receptors (DR4/DR5) and enhanced via intrinsic pathway by modulating Bcl‐2 and inhibitor of apoptosis protein family members. These are related to ROS generation. In conclusion, this study provides evidence that morin can enhance the anticancer activity of AF in Hep3B human hepatocellular carcinoma cells, indicating that its combination could be an alternative treatment strategy for the hepatocellular carcinoma.     

Corosolic Acid Triggers Mitochondria and Caspase‐dependent Apoptotic Cell Death in Osteosarcoma MG‐63 Cells

The response of osteosarcoma MG‐63 cells to corosolic acid treatment has been investigated. The results showed that corosolic acid significantly inhibited cell viability in both a dose and a time dependent manner. It was found that corosolic acid increased the Bax/Bcl‐2 ratio by up‐regulating Bax expression, disrupted mitochondrial membrane potential and triggered the release of cytochrome c from mitochondria into the cytoplasm. Corosolic acid treatment triggered the activation of caspase‐8, 9 and 3. The apoptosis was obviously inhibited by pretreatment with a general caspase inhibitor, z‐VAD‐FMK. Moreover, pretreatment of CsA, a cyclophilin D ligand that inhibits mitochondria potential uncoupling, prevented the activation of caspase‐9 and caspase‐3, but not caspase‐8, and the apoptosis of MG‐63 cells, triggered by corosolic acid. All these results indicated that corosolic acid‐induced apoptosis was associated with the activation of caspases via a mitochondrial pathway. Copyright © 2011 John Wiley & Sons, Ltd.     

Ramalin‐Mediated Apoptosis Is Enhanced by Autophagy Inhibition in Human Breast Cancer Cells

Breast cancer, the most commonly diagnosed cancer in women worldwide, is treated in various ways. Ramalin is a chemical compound derived from the Antarctic lichen Ramalina terebrata and is known to exhibit antioxidant and antiinflammatory activities. However, its effect on breast cancer cells remains unknown. We examined the ability of ramalin to induce apoptosis and its mechanisms in MCF‐7 and MDA‐MB‐231 human breast cancer cell lines. Ramalin inhibited cell growth and induced apoptosis in both cell lines in a concentration‐dependent manner. By upregulating Bax and downregulating Bcl‐2, ramalin caused cytochrome c and apoptosis‐inducing factor to be released from the mitochondria into the cytosol, thus activating the mitochondrial apoptotic pathway. In addition, activated caspase‐8 and caspase‐9 were detected in both types of cells exposed to ramalin, whereas ramalin activated caspase‐3 only in the MDA‐MB‐231 cells. Ramalin treatment also increased the levels of LC3‐II and p62. Moreover, the inhibition of autophagy by 3‐methyladenine or Atg5 siRNA significantly enhanced ramalin‐induced apoptosis, which was accompanied by a decrease in Bcl‐2 levels and an increase in Bax levels. Therefore, autophagy appears to be activated as a protective mechanism against apoptosis in cancer cells exposed to ramalin. These findings suggest that ramalin is a potential anticancer agent for the treatment of patients with non‐invasive or invasive breast cancer. Copyright © 2015 John Wiley & Sons, Ltd.     

Mangosenone F,A Furanoxanthone from Garciana mangostana,Induces Reactive Oxygen Species‐Mediated Apoptosis in Lung Cancer Cells and Decreases Xenograft Tumor Growth

Mangosenone F (MSF), a natural xanthone, was isolated form Carcinia mangotana, and a few studies have reported its glycosidase inhibitor effect. In this study we investigated the anti lung cancer effect of MSF both in vitro and in vivo. MSF inhibited cancer cell cytotoxicity and induced and induced apoptosis via reactive oxygen species (ROS) generation in NCI‐H460. MSF treatment also showed in pronounced release of apoptogenic cytochrome c from the mitochondria to the cytosol, downregulation of Bcl‐2 and Bcl‐xL, and upregulation of Bax, suggesting that caspase‐mediated pathways were involved in MSF‐induced apoptosis. ROS activation of the mitogen‐activated protein kinase signaling pathway was shown to play a predominant role in the apoptosis mechanism of MSF. Compared with cisplatin treatment, MSF treatment showed significantly increased inhibition of the growth of NCI‐H460 cells xenografted in nude mice. Together, these results indicate the potential of MSF as a candidate natural anticancer drug by promoting ROS production. Copyright © 2015 John Wiley & Sons, Ltd.