β‐Eudesmol Induces JNK‐Dependent Apoptosis through the Mitochondrial Pathway in HL60 Cells

β‐eudesmol, a natural sesquiterpenol present in a variety of Chinese herbs, is known to inhibit the proliferation of human tumor cells. However, the molecular mechanisms of the effect of β‐eudesmol on human tumor cells are unknown. In the present study, we report the cytotoxic effect of β‐eudesmol on the human leukemia HL60 cells and its molecular mechanisms. The cytotoxic effect of β‐eudesmol on HL60 cells was associated with apoptosis, which was characterized by the presence of DNA fragmentation. β‐eudesmol‐induced apoptosis was accompanied by cleavage of caspase‐3, caspase‐9, and poly (ADP‐ribose) polymerase; downregulation of Bcl‐2 expression; release of cytochrome c from mitochondria; and decrease in mitochondrial membrane potential (MMP). Activation of c‐Jun N‐terminal kinases (JNK) mitogen‐activated protein kinases was observed in β‐eudesmol‐treated HL60 cells, and the inhibitor of JNK blocked the β‐eudesmol‐induced apoptosis, downregulation of Bcl‐2, and the loss of MMP. These data suggest that β‐eudesmol induces apoptosis in HL60 cells via the mitochondrial apoptotic pathway, which is controlled through JNK signaling. Copyright © 2012 John Wiley & Sons, Ltd.     

Chebulinic Acid Isolated From the Fruits of Terminalia chebula Specifically Induces Apoptosis in Acute Myeloid Leukemia Cells

Chebulinic acid, an ellagitannin found in the fruits of Terminalia chebula, has been extensively used in traditional Indian system of medicine. It has shown to have various biological activities including antitumor activity. The present study aims to investigate the cytotoxic potential of chebulinic acid in human myeloid leukemia cells. Interestingly, chebulinic acid caused apoptosis of acute promyelocytic leukemia HL‐60 and NB4 cells but not K562 cells. In vitro antitumor effects of chebulinic acid were investigated by using various acute myeloid leukemia cell lines. Chebulinic acid treatment to HL‐60 and NB4 cells induced caspase activation, cleavage of poly(ADP‐ribose) polymerase, DNA fragmentation, chromatin condensation, and changes in the mitochondrial membrane permeability. Additionally, inhibition of caspase activation drastically reduced the chebulinic acid‐induced apoptosis of acute promyelocytic leukemia cells. Our data also demonstrate that chebulinic acid‐induced apoptosis in HL‐60 and NB4 cells involves activation of extracellular signal‐regulated kinases, which, when inhibited with ERK inhibitor PD98059, mitigates the chebulinic acid‐induced apoptosis. Taken together, our findings exhibit the selective potentiation of chebulinic acid‐induced apoptosis in acute promyelocytic leukemia cells. Copyright © 2017 John Wiley & Sons, Ltd.     

Aloe‐emodin Induces Apoptosis in Human Liver HL‐7702 Cells through Fas Death Pathway and the Mitochondrial Pathway by Generating Reactive Oxygen Species

Aloe‐emodin (1,8‐dihydroxy‐3‐hydroxymethyl‐anthraquinone) is one of the primary active compounds in total rhubarb anthraquinones isolated from some traditional medicinal plants such as Rheum palmatum L. and Cassia occidentalis, which induce hepatotoxicity in rats. Thus, the aim of this study was to determine the potential cytotoxic effects and the underlying mechanism of aloe‐emodin on human normal liver HL‐7702 cells. The CCK‐8 assays demonstrated that aloe‐emodin decreased the viability of HL‐7702 cells in a dose‐dependent and time‐dependent manner. Aloe‐emodin induced S and G2/M phase cell cycle arrest in HL‐7702 cells. This apoptosis was further investigated by flow cytometry and nuclear morphological changes by DAPI staining, respectively. Moreover, aloe‐emodin provoked the production of intracellular reactive oxygen species and the depolarization of mitochondrial membrane potential (MMP). Further studies by western blot indicated that aloe‐emodin dose‐dependently up‐regulated the levels of Fas, p53, p21, Bax/Bcl‐2 ratio, and cleaved caspase‐3, ‐8, ‐9, and subsequent cleavage of poly(ADP‐ribose)polymerase (PARP). Taken together, these results suggest that aloe‐emodin inhibits cell proliferation of HL‐7702 cells and induces cell cycle arrest and caspase‐dependent apoptosis via both Fas death pathway and the mitochondrial pathway by generating reactive oxygen species, indicating that aloe‐emodin should be taken into account in the risk assessment for human exposure. Copyright © 2017 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.     

Emodin Regulates Apoptotic Pathway in Human Liver Cancer Cells

Emodin, a natural anthraquinone, has been reported to possess antiproliferative effects in many cancer cell lines. However, anticancer mechanism against human liver cancer remains unclear. In this study, we observed that emodin induced apoptosis in HepG2 cells and caused a significant accumulation of cells in the G1 phase. Western blot data showed that emodin treatment caused the increasing of release of cytochrome c into cytosol from mitochondria and the activation of caspase‐8 and caspase‐9, which suggest that the intrinsic and extrinsic pathways could be involved. Emodin treatment also resulted in a dose‐dependent accumulation of intracellular reactive oxygen species. Furthermore, emodin increased the protein level of p53 and decreased the protein level of NF‐κB/p65 in HepG2 cells, which indicated these two regulators might play a role in emodin‐induced apoptosis. Computational modeling showed that emodin could directly bind to the BH3 domain of Bcl‐2 through forming one hydrogen bond with Ala146 residue in Bcl‐2. From these examinations, emodin not only significantly downregulated expression of Bcl‐2 but also inhibited the heterodimerization of Bcl‐2 with Bax because of strong interaction between emodin and Bcl‐2. These suggest that emodin induces apoptosis in liver cancer cell line through a multifaceted complex cascade of events. Copyright © 2012 John Wiley & Sons, Ltd.     

Anticancer activity and molecular mechanisms of α‐conidendrin,a polyphenolic compound present in Taxus yunnanensis,on human breast cancer cell lines

α‐Conidendrin is a polyphenolic compound found mainly in Taxus yunnanensis, as the source of chemotherapy drug paclitaxel, which has been used in traditional medicine for treatment of cancer. This study aimed to investigate the anticancer activity and molecular mechanisms of α‐conidendrin on breast cancer cell lines. The results of the present study show that α‐conidendrin possesses potent antiproliferative effects on breast cancer cell lines MCF‐7 and MDA‐MB‐231. α‐Conidendrin significantly induced apoptosis in breast cancer cells via reactive oxygen species generation, upregulation of p53 and Bax, downregulation of Bcl‐2, depolarization of mitochondrial membrane potential (MMP), release of cytochrome c from mitochondria, and activation of caspases‐3 and ‐9. α‐Conidendrin remarkably inhibited the proliferation of breast cancer cells through induction of cell cycle arrest by upregulating p53 and p21 and downregulating cyclin D1 and CDK4. Unlike breast cancer cells, the antiproliferative effect of α‐conidendrin on human foreskin fibroblast cells (normal cells) was very small. In normal cells, reactive oxygen species levels, loss of MMP, release of cytochrome c, mRNA expression of p53, p21, cyclin D1, CDK4, Bax, and Bcl‐2 as well as mRNA expression and activity of caspases‐3 and ‐9 were significantly less affected by α‐conidendrin compared with cancer cells. These results suggest that α‐conidendrin can be a promising agent for treatment of breast cancer with little or no toxicity against normal cells.     

Cardioprotective Effects of Tannic Acid on Isoproterenol‐Induced Myocardial Injury in Rats: Further Insight into ‘French Paradox’

Tannic acid (TA) is a polyphenolic compound, which has shown diverse pharmacological effects with antimutagenic, anticarcinogenic and antibactericidal properties. However, cardioprotective effects of TA have not been reported. To investigate the protective effects of TA, rats were administered TA for 7 days and then intoxicated with isoproterenol (ISO). Myocardial ischemia injury was indicated by changes in electrocardiographic (ECG) patterns, morphology and cardiac marker enzymes. Furthermore, protein expression levels of c‐fos, c‐jun, tumor necrosis factor‐α (TNF‐α), interleukin‐1β (IL‐1β), cleaved‐caspase‐3 and ‐9 were analyzed by immunohistochemistry, and activities of apoptosis‐related proteins Bax, Bcl‐2, caspase‐3 and nuclear factor kappa B (NF‐κB) were detected by Western blot. Pretreatment with TA ameliorated changes in morphology and ECG, reduced activities of marker enzymes, suppressed overexpression of apoptosis‐related proteins, upregulated expression of antioxidants. Moreover, TA pretreatment contributed to the decrease in ratio of Bax/Bcl‐2, as well as reduced expression of TNF‐α, IL‐1β, caspase‐3, cleaved‐caspase‐3 and ‐9. TA displayed cardioprotective effects, which may be attributed to lowering of Bax/Bcl‐2 ratio, c‐fos and c‐jun expression and inhibition of NF‐κB activation, as well as oxidative stress, inflammation and apoptosis. These findings provide further insight into the ‘French paradox’ and the mechanisms underlying the beneficial effects of TA. Copyright © 2015 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.     

Antitumour effects of selected plant polyphenols,gallic acid and ellagic acid,on sensitive and multidrug‐resistant leukaemia HL60 cells

The aim of this study was to examine the antitumour effects of plant phenolic acids, gallic acid (GA) and ellagic acid (EA), on human promyelocytic leukaemia sensitive HL60 cell line and its resistant sublines exhibiting two MDR phenotypes: HL60/VINC (overexpressing P‐glycoprotein) and HL60/MX2 (characterized by the presence of mutated α isoform of topoisomerase II). Both studied compounds exerted comparable cytotoxic activities towards sensitive HL60 cells and their MDR counterparts. It was also found that GA and EA modulated the cellular level of reactive oxygen species in a dose‐dependent and time‐dependent manner. Furthermore, it was demonstrated that GA (IC₉₀) and EA (IC₅₀ and IC₉₀) significantly increased the percentage of sub‐G1 subpopulation of all studied leukaemia cells causing oligonucleosomal DNA fragmentation. Both compounds used at IC₉₀ triggered mainly the apoptotic death of these cells. However, GA had no effect on the activity of caspase‐3 as well as caspase‐8 in sensitive HL60 cells and their MDR counterparts. In contrast, EA provoked a significant activation of these caspases in all studied leukaemia cells. It was also found that lysosomes were not involved in triggering programmed death of sensitive HL60 and MDR cells by GA and EA.     

丹参酮ⅡA诱导白血病细胞凋亡过程中端粒酶活性的改变

目的观察丹参酮Ⅱ_A(TanⅡ_A)对HL60,K562细胞端粒酶活性的抑制作用和对凋亡相关基因的影响,探讨丹参酮Ⅱ_A对造血细胞的作用机理。方法以HL60,K562为靶细胞,应用细胞培养技术,流式细胞术,透射电镜观察TanⅡ_A对HL60,K562细胞的作用,利用PCRTRAP方法检测TanⅡ_A处理前后HL60,K562细胞端粒酶活性的改变。结果经05μg·mL^-1TanⅡ_A作用6d后,HL60,K562细胞生长明显受到抑制,生长抑制率分别为756%和563%。经丹参酮诱导后,HL60,K562细胞发生凋亡,出现亚二倍体峰;同时显著下调HL60及K562细胞的cmyc,bcl2基因表达,上调cfos基因表达。HL60,K562细胞在TanⅡ_A作用后,端粒酶活性受到抑制,端粒酶活性抑制率分别为308%,508%。结论TanⅡ_A可明显抑制HL60和K562细胞的增殖和细胞端粒酶活性,并诱导细胞凋亡。     

(−)‐Epigallocatechin‐3‐Gallate Ameliorates Photodynamic Therapy Responses in an In Vitro T Lymphocyte Model

(?)‐Epigallocatechin‐3‐gallate (EGCG), the most abundant polyphenolic constituent in green tea, is known as a powerful antioxidant but concomitantly possesses a prooxidant property. We investigated the effect of EGCG on phloxine B (PhB)‐induced photocytotoxicity in human T lymphocytic leukemia Jurkat cells. EGCG significantly potentiated PhB‐induced photocytotoxic effects, including the inhibition of cell proliferation, DNA fragmentation, and caspase‐3 activity induction in Jurkat cells. Catalase attenuated the enhanced cytotoxicity by EGCG, suggesting the involvement of extracellularly produced hydrogen peroxide. Indeed, EGCG significantly enhanced extracellular hydrogen peroxide formation induced by photo‐irradiated PhB. The EGCG also enhanced intracellular reactive oxygen species accumulation, c‐Jun N ‐terminal kinase (JNK) phosphorylation, and interferon‐γ (IFN‐γ) gene expression, all of which are involved in PhB‐induced apoptosis. Taken together, our data suggest that EGCG is capable of potentiating photodynamic therapy responses, presumably through the intracellular oxidative stress‐sensitive JNK/IFN‐γ pathway by exogenous hydrogen peroxide formation. Copyright © 2014 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.     

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.     

粟米草三萜类化学成分及其活性研究

目的研究粟米草Mollugo pentaphylla中的三萜类化学成分,寻找其中具有细胞毒活性化合物。方法综合运用大孔吸附树脂、硅胶柱色谱、半制备型高效液相及SephadexLH-20凝胶柱色谱等各种色谱技术进行系统化学研究,根据其理化性质和MS、NMR等波谱数据鉴定化合物结构,同时对所得的三萜皂苷进行细胞毒活性测试。结果从粟米草干燥地上部分醇提取物中共分离得到6个三萜类化合物,分别鉴定为粟米草苷E(1)、3-O-[α-L-rhamnopyranosy1(1→2)-α-L-arabinopyranosyl]-28-O-[β-D-glucopyranosyl (1→6)-β-D-glucopyranosyl] oleanolic acid(2)、竹节香附皂苷R8(3)、竹节香附素A(4)、mollugogenolsA(5)、齐墩果酸(6)。细胞毒活性显示,化合物1~5对人前列腺癌DU145细胞、人宫颈癌HeLa细胞及人早幼粒白血病HL-60细胞均显示一定的抑制作用,尤其是对人早幼粒急性白血病HL-60细胞,其IC50分别为10.21、38.43、40.28、20.59、83.16μmol/L。结论化合物1为新的齐墩果酸型三萜皂苷,2~4为首次从该植物中分离得到。细胞毒活性显示,化合物1~5对人前列腺癌DU145细胞、人宫颈癌HeLa细胞及人早幼粒白血病HL-60细胞均显示一定的抑制作用。     

Antiproliferative Activity of Phenylpropanoids Isolated from Lagotis brevituba Maxim

The aim of the present study was to evaluate the antiproliferative effect of phenylpropanoids isolated from the n‐BuOH‐soluble fraction of an ethanolic extract of Lagotis brevituba Maxim. The phenylpropanoids were identified as echinacoside, lagotioside, glucopyranosyl(1–6)martynoside, plantamoside, and verbascoside. Three of the compounds, lagotioside, glucopyranosyl(1–6)martynoside, and plantamoside, were isolated from L. brevituba for the first time. The antiproliferative activity of the isolates was evaluated in human gastric carcinoma (MGC‐803), human colorectal carcinoma (HCT116), human hepatocellar carcinoma (HepG2), and human lung cancer (HCT116) cells using an 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay. Plantamoside showed promising activity against MGC‐803 cells, with a half maximal inhibitory concentration value of 37.09 μM. The mechanism of the pro‐apoptosis effect of plantamoside was then evaluated in MGC‐803 cells. Changes in cell morphology, including disorganization of the architecture of actin microfilaments and formation of apoptotic bodies, together with cell cycle arrest in G2/M phases, were observed after treatment of plantamoside. The antiproliferative and pro‐apoptotic effects were associated with a decrease in the ratio of Bcl‐2/Bax and reduced mitochondrial membrane potential, which was accompanied by the release of reactive oxygen species and Ca²⁺ into the cytoplasm. Taken together, the results indicated that plantamoside promotes apoptosis via a mitochondria‐dependent mechanism. Copyright © 2017 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.     

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.     

Antiproliferative effect of flavonoids and sesquiterpenoids from Achillea millefolium s.l. on cultured human tumour cell lines

The antiproliferative activities of n‐hexane, chloroform, aqueous‐methanol and aqueous extracts of the aerial parts of the Achillea millefolium aggregate on three human tumour cell lines were investigated by means of MTT assays. The chloroform‐soluble extract exerted high tumour cell proliferation inhibitory activities on HeLa and MCF‐7 cells, and a moderate effect on A431 cells; accordingly, it was subjected to detailed bioactivity‐guided fractionation. As a result of the multistep chromatographic purifications (VLC, CPC, PLC, gel filtration), five flavonoids (apigenin, luteolin, centaureidin, casticin and artemetin) and five sesquiterpenoids (paulitin, isopaulitin, psilostachyin C, desacetylmatricarin and sintenin) were isolated and identified by spectroscopic methods. The antiproliferative assay demonstrated that centaureidin is the most effective constituent of the aerial parts of yarrow: high cell growth inhibitory activities were observed especially on HeLa (IC₅₀ 0.0819 µm ) and MCF‐7 (IC₅₀ 0.1250 µm ) cells. Casticin and paulitin were also highly effective against all three tumour cell lines (IC₅₀ 1.286–4.76 µm ), while apigenin, luteolin and isopaulitin proved to be moderately active (IC₅₀ 6.95–32.88 µm ). Artemetin, psilostachyin C, desacetylmatricarin and sintenin did not display antiproliferative effects against these cell lines. This is the first report on the occurrence of seco‐pseudoguaianolides (paulitin, isopaulitin and psilostachyin C) in the Achillea genus. Copyright © 2008 John Wiley & Sons, Ltd.     

A p‐Menth‐1‐ene‐4,7‐diol (EC‐1) from Eucalyptus camaldulensis Dhnh. Triggers Apoptosis and Cell Cycle Changes in Ehrlich Ascites Carcinoma Cells

Anticancer activities of p‐menth‐1‐ene‐4,7‐diol (EC‐1) isolated from Eucalyptus camaldulensis Dhnh. were studied on Ehrlich ascites carcinoma (EAC) cells by MTT (3‐[4,5‐dimethylthiazol‐2‐yl]‐2,5 diphenyl tetrazolium bromide) assay. Anticancer activities also analyzed in EAC‐bearing mice by assessment of cancer growth inhibition, changes in cancer volume, changes in life span, and hematological parameters. Apoptosis was analyzed by fluorescence microscope, DNA fragmentation assay, and flow cytometry. The expression of apoptosis‐related genes, Bcl‐2, Bcl‐X, PARP‐1, p53, and Bax, were analyzed using polymerase chain reaction (PCR). EC‐1 significantly inhibited proliferation of EAC cells in vivo and restored the altered hematological parameters of EAC‐bearing mice. Cytological observation by fluorescence microscope showed apoptosis of EAC cells upon treatment with EC‐1. Also, DNA fragmentation assay revealed EAC cells' apoptosis following EC‐1 treatment. Increased mRNA expressions of p53 and Bax genes and negative expressions of Bcl‐2 and Bcl‐X were observed in cells treated with EC‐1. These findings confirmed the induction of apoptosis by EC‐1. In addition, MTT assay showed dose‐dependent anticancer activity of EC‐1 against EAC cell. Cell cycle analysis revealed that EC‐1 treatment caused suppression of EAC cells at S phase. To conclude, EC‐1 is a novel anticancer compound and showed antiproliferative and apoptotic activities in cellular and mice models. Copyright © 2015 John Wiley & Sons, Ltd.