Ethanolic extract of Descurainia sophia seeds sensitizes A549 human lung cancer cells to TRAIL cytotoxicity by upregulating death receptors
Chae Jun Lim,
Ok-Sun Bang and
No Soo Kim
Abstract Background Our previous genome-wide gene expression analysis revealed that tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptors 4 (DR4) and 5 (DR5) are markedly upregulated by the ethanolic extract of D. sohia seeds (EEDS) in A549 TRAIL-refractory cancer cells. In the present study, we investigated whether the EEDS-mediated upregulation of TRAIL death receptors was associated with increased TRAIL-mediated toxicity in A549 cells in vitro.
Methods Cell proliferation and viability were determined by an automatic cell counter. Gene silencing was performed by introducing small interfering RNA into cells. Expression changes of cellular proteins were determined by western blot analysis. Apoptotic cell death was monitored by western blot analysis. Analysis of variance followed by the post-hoc Dunnett’s test was used to compare the data.
Results EEDS treatment increased both mRNA and protein levels of DR4 and DR5 in the TRAIL refractory A549 cells. Co-treatment of A549 cells with sub-lethal dose of EEDS and recombinant TRAIL increased the apoptotic cell death. Upregulation of DR5 by EEDS was mediated by an endoplasmic reticulum stress-induced transcription factor, CCAAT/enhancer-binding protein homologous protein (CHOP), and knockdown of CHOP expression inhibited EEDS-induced DR5 upregulation and abolished the EEDS-associated increase in TRAIL toxicity in A549 cells.
Conclusions EEDS can sensitize A549 cells to TRAIL cytotoxicity by upregulation of TRAIL death receptors. Our findings suggested that EEDS is a good initial herbal source for the development of an anticancer supplement for anticancer therapeutics associated with TRAIL.
Isolation of methyl gamma linolenate from spirulina platensis using flash chromatography and its apoptosis inducing effect Jubie S.1, Dhanabal S. P.2* and Chaitanya M. V. N. L.
Abstract Background Isolation of methyl gamma linolenate from Spirulina platensis using flash chromatography and its apoptosis inducing effect against human lung carcinoma A- 549 cell lines.
Methods Gamma linolenic acid is an important omega-6 polyunsaturated fatty acid (PUFA) of medicinal interest was isolated from microalgae Spirulina platensis using flash chromatography system (Isolera system) as its methyl ester. The isolated methyl gamma linolenate was characterized by IR, 1 H NMR, 13 C NMR and mass spectral analysis and the data were consistent with the structure.
Results The percentage yield of isolated methyl gamma linolenate is found to be 71 % w/w, which is a very good yield in comparison to other conventional methods. It was subjected to in-vitro cytotoxic screening on A-549 lung cancer cell lines using SRB assay and result was compared with standard rutin.
Conclusion It may be concluded that the Flash chromatography system plays a major role in improving the yield for theisolation of methyl gamma linoleate from Spirulina platensis and the isolated molecule is a potent cytotoxicagent towards human lung carcinoma cell lines, however it may be further taken up for an extensive study.
Anti-Inflammatory and Pro-apoptotic Effects of Curcumin and Resveratrol on the Human Lung Fibroblast Cell Line MRC-5 Burkhard Kloesch1*, Elisabeth Dietersdorfer1, Silvia Loebsch1 and Guenter Steiner1,2
1Ludwig Boltzmann Institute for Rheumatology and Balneology, Cluster Rheumatology, Balneology and Rehabilitation, Vienna, Austria 2Division of Rheumatology, Department of Internal Medicine III, Medical University Vienna, Austria
Abstract Background: The naturally occuring polyphenols curcumin and resveratrol are considered to be powerful antioxidants and anti-inflammatory compounds and both inhibit the proliferation of different types of cancer cells. In the present study, we investigated possible anti-inflammatory and pro-apoptotic effects of curcumin and resveratrol on the human lung fibroblast cell line MRC-5.
Methods: MRC-5 cells were stimulated for 6 h with interleukin (IL)-1β or phorbol 12- myristate 13-acetate (PMA) in the absence or presence of different concentrations of curcumin or resveratrol. The release of interleukin (IL)-6 was quantified by enzyme-linked immunosorbent assay (ELISA). The modulation in phosphorylation of the transcription factor nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) such as p38 and ERK1/2 were analyzed by Western blot. Cytotoxic and pro-apoptotic effects of curcumin and resveratrol were monitored by the measurement of lactate dehydrogenase (LDH) activity and by Annexin-V/7-AAD staining.
Results: Both curcumin and resveratrol effectively attenuated IL-1β and PMA-induced IL-6 expression in MRC-5 cells. Furthermore, curcumin treatment induced apoptosis via caspase-3 signaling and caused endoplasmic reticulum (ER) stress. Salubrinal, an inhibitor of serine/threonine phosphatase PP1, and antioxidants such as N-acetyl-cysteine (NAC), reduced glutathione (GSH) and sodium hydrogen sulfide (NaHS) diminished the cytotoxic effects of curcumin on MRC-5 cells. In contrast to curcumin, resveratrol had no negative effects on cell viability
Abstract HLJ1 is a novel tumour suppressor and is a potential druggable target for non-small-cell lung cancer (NSCLC). In this report, using a promoter-containing enhancer region as the HLJ1-targeting drug-screening platform, we identified several herbal compounds from a Chinese herbal bank with the capacity to enhance HLJ1 promoter activity and suppress tumour growth and invasion of NSCLC. Among the herbal drugs identified, the andrographolide (from Andrographis paniculata [Burm. f.] Nees.) most significantly induced HLJ1 expression and suppressed tumorigenesis both in vitro and in vivo. The andrographolide upregulates HLJ1 via JunB activation, which modulates AP-2α binding at the MMP-2 promoter and represses the expression of MMP-2. In addition, silencing of HLJ1 partially reverses the inhibition of cancer-cell invasion by andrographolide. Microarray transcriptomic analysis was performed to comprehensively depict the andrographolide-regulated signalling pathways. We showed that andrographolide can affect 939 genes (analysis of variance, false discovery rate < 0.05) that are dominantly involved in the cell cycle, apoptosis and adhesion-related biological signalling, including mitogen-activated protein kinase, focal adhesion and tight junction pathways, indicating the diverse effects of andrographolide on anticancer invasion and proliferation. In conclusion, the HLJ1-targeting drug-screening platform is useful for screening of novel anticancer compounds. Using this platform, we identified andrographolide is a promising new anticancer agent that could suppress tumour growth and invasion in NSCLC.
Gambogic acid synergistically potentiates cisplatin-induced apoptosis in non-small-cell lung cancer through suppressing NF-κB and MAPK/HO-1 signalling L-H Wang1,2, Y Li1, S-N Yang1, F-Y Wang1, Y Hou1, W Cui1, K Chen1, Q Cao1, S Wang1, T-Y Zhang1, Z-Z Wang2, W Xiao2, J-Y Yang1 and C-F Wu1
1Department of Pharmacology, Shenyang Pharmaceutical University, Shenyang 110016, People’s Republic of China
2Jiangsu Kanion Pharmaceutical Co. Ltd, Lianyungang 222001, People’s Republic of China
Abstract Background: Gambogic acid (GA) has been reported to have potent anticancer activity and is authorised to be tested in phase II clinical trials for treatment of non-small-cell lung cancer (NSCLC). The present study aims to investigate whether GA would be synergistic with cisplatin (CDDP) against the NSCLC.
Methods: 1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT), combination index (CI) isobologram, western blot, quantitative PCR, flow cytometry, electrophoretic mobility shift assay, xenograft tumour models and terminal deoxynucleotide transferase-mediated dUTP nick-end labelling analysis were used in this study.
Results: The cell viability results showed that sequential CDDP-GA treatment resulted in a strong synergistic action in A549, NCI-H460, and NCI-H1299 cell lines, whereas the reverse sequence and simultaneous treatments led to a slight synergistic or additive action. Increased sub-G1 phase cells and enhanced PARP cleavage demonstrated that the sequence of CDDP-GA treatment markedly increased apoptosis in comparison with other treatments. Furthermore, the sequential combination could enhance the activation of caspase-3, -8, and 9, increase the expression of Fas and Bax, and decrease the expression of Bcl-2, survivin and X-inhibitor of apoptosis protein (X-IAP) in A549 and NCI-H460 cell lines. In addition, increased apoptosis was correlated with enhanced reactive oxygen species generation. Importantly, it was found that, followed by CDDP treatment, GA could inhibit NF-κB and mitogen-activated protein kinase (MAPK)/heme oxygenase-1 (HO-1) signalling pathways, which have been validated to reduce ROS release and confer CDDP resistance. The roles of NF-κB and MAPK pathways were further confirmed by using specific inhibitors, which significantly increased ROS release and apoptosis induced by the sequential combination of CDDP and GA. Moreover, our results indicated that the combination of CDDP and GA exerted increased antitumour effects on A549 xenograft models through inhibiting NF-κB, HO-1, and subsequently inducing apoptosis.
Conclusion: Gambogic acid sensitises lung cancer cells to CDDP in vitro and in vivo in NSCLC through inactivation of NF-κB and MAPK/HO-1 signalling pathways, providing a rationale for the combined use of CDDP and GA in lung cancer chemotherapy.
Antineoplastic effects of deoxyelephantopin, a sesquiterpene lactone from Elephantopus scaber, on lung adenocarcinoma (A549) cells Farha A. Kabeer1, Geetha B. Sreedevi2, Mangalam S. Nair3, Dhanya S. Rajalekshmi3, Latha P. Gopalakrishnan2, Sujathan Kunjuraman1, Remani Prathapan1
1.Division of Cancer Research, Regional Cancer Centre, Thiruvananthapuram, Pin 695011, Kerala, India 2.Jawaharlal Nehru Tropical Botanic Garden and Research Institute, Pin 695562, Thiruvananthapuram, India 3.National Institute of Interdisciplinary Science and Technology, Pin 695019, Thiruvananthapuram, India
Abstract OBJECTIVE: Deoxyelephantopin, a sesquiterpene lactone from Elephantopus scaber, showed inhibition of the growth of various tumor cells in vitro. In the present study, we investigated the cytotoxicity and apoptosis-inducing capacity of deoxyelephantopin on lung adenocarcinoma (A549) cells.
METHODS: The cytotoxic effect of deoxyelephantopin on A549 cells and normal lymphocytes was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 50% inhibitory concentration (IC50) value was determined. The self-renewal and proliferating potential of A549 cells after treatment with deoxyelephantopin were examined by colony formation assay. Cellular morphology of deoxyelephantopin-treated cells was observed using phase-contrast microscopy. The induction of apoptosis was evaluated using acridine orange and ethidium bromide staining, Hoechst 33342 staining, terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end-labeling (TUNEL) assay, DNA fragmentation analysis and Annexin V-fluorescein isothiocyanate staining by flow cytometry. Activation of caspases was detected using fluorogenic substrate specific to caspases 2, 3, 8 and 9 and flow cytometric analysis. The total cellular DNA content and expression of cleaved poly (ADP-ribose) polymerase was also analyzed.
RESULTS: Deoxyelephantopin exhibited cytotoxicity to A549 cells (IC50 = 12.287 μg/mL), however, there was no toxicity towards normal human lymphocytes. Deoxyelephantopin suppressed the colony-forming ability of A549 cells in a dose-dependent manner. Acridine orange, ethidium bromide and Hoechst 33342 staining showed cell shrinkage, chromosomal condensation and nuclear fragmentation, indicating induction of apoptosis. Deoxyelephantopin increased apoptosis of A549 cells, as evidenced by more TUNEL-positive cells. DNA fragmentation and Annexin V staining revealed late-stage apoptotic cell population. Deoxyelephantopin inhibited A549 cell growth by cell cycle arrest at G2/M phase and induced apoptosis through both extrinsic and intrinsic pathways.
CONCLUSION: These results suggest that deoxyelephantopin has great potential as a new chemotherapeutic agent to be developed further for the treatment of lung cancer
Anticancer activity of botanical compounds in ancient fermented beverages (Review) P.E. McGOVERN1, M. CHRISTOFIDOU-SOLOMIDOU2,3, W. WANG4, F. DUKES2, T. DAVIDSON1 and W.S. EL-DEIRY4
1Biomolecular Archaeology Laboratory, University of Pennsylvania Museum of Archaeology and Anthropology; 2Pulmonary, Allergy and Critical Care Division, 3Center of Excellence in Environmental Toxicology (CEET), and 4Division of Hematology and Oncology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA
Abstract Humans around the globe probably discovered natural remedies against disease and cancer by trial and error over the millennia. Biomolecular archaeological analyses of ancient organics, especially plants dissolved or decocted as fermented beverages, have begun to reveal the preliterate histories of traditional pharmacopeias, which often date back thousands of years earlier than ancient textual, ethnohistorical, and ethnological evidence. In this new approach to drug discovery, two case studies from ancient Egypt and China illustrate how ancient medicines can be reconstructed from chemical and archaeological data and their active compounds delimited for testing their anticancer and other medicinal effects. Specifically, isoscopoletin from Artemisia argyi, artemisinin from Artemisia annua, and the latter's more easily assimilated semi-synthethic derivative, artesunate, showed the greatest activity in vitro against lung and colon cancers. In vivo tests of these compounds previously unscreened against lung and pancreatic cancers are planned for the future.
Conclusion ..These data show that both artesunate and artemisinin treatments resulted in a concentration-dependent inhibition of cell growth in Lewis lung carcinoma cells, but that artesunate was >30 times as effective ... ...The active compounds detected by the p53 studies included artemisinin, artesunate, borneol, isoscopoletin, and ursolic acid, with inhibition effects on tumor growth ranging in concentration from micromolar to millimolar amounts. Although artesunate, and to a lesser extent artemisinin, are most active against HCT116 colon adenocarcinoma... ...ursolic acid, found in species of thyme, is much more effective against colon cancer under low-oxygen conditions (hypoxia) than normal atmospheric conditions (normoxia). This compound inhibits tumor growth at a concentration of around 50 μM under normoxia; much lower concentrations of 1-10 μM are sufficient to achieve the same result under hypoxia. Tumor cells, especially those which are solid and dense, survive and proliferate under hypoxia. A strategy in anticancer therapy is to counteract this property of tumor cells. It may be hypothesized that the anticancer potency of ursolic acid is related to mechanisms operating under hypoxia... ...Based on these results, we propose in vivo and clinical studies of one derivative compound from A. annua in particular: artesunate. As already known from in vivo mouse studies of other cancers, we plan to test this compound for its clinical efficacy against lung and pancreatic cancer. To date, the effectiveness of artesunate against these specific cancers has not been determined...
Aloe-emodin novel anticancer Herbal Drug Khemkaran Ahirwar1 * Sanmati K. Jain1
Abstract The electrochemical behaviour of the anticancer herbal drug emodin hydroxyanthraquinone present in Aloe vera leaves has a specific in vitro and in vivo antineuroectodermal tumor activity. The compound does not inhibit the proliferation of normal fibroblasts n or that of hemopoietic progenitor cells. The cytotoxicity mechanism consists of the induction of apoptosis, whereas the selectivity against neuroectodermal tumor cells is founded on a specific energy-dependent pathway of drug incorporation. Natural compounds that have traditionally been used to treat a variety of diseases for hundreds of years (1, 2, 3) . We assayed only those natural compounds that have already been proven to be nontoxic, and we evaluated their efficacy against highly malignant tumors that are not normally included in the classical screening assays.
Conclusion: Aloe-Emodin was able to inhibit cell growth in several tumor cells, including human lung carcinoma, 10 hepatoma, 11 and leukemia cell lines. 13 aloe-emodin shows a high specificity for neuroectodermal tumor cells. 14 one of the important approaches for cancer chemotherapy is to regulate cell-cycle progression. G1/S cell-cycle arrest was found in human hepatoma, 12 glioma, 16 breast, 10 lung, 11 and colon 15 carcinoma cells upon treatment of rhubarb anthraquinones (emodin, 9 aloe-emodin, 13 and rhein 16 The herbal medicines have great importance in the treatment of many diseases. Since herbal medicines are mainly used by Chinese, but now gaining acceptance all over the world and mostly in India. Herbal plants and their derivatives are widely used in the treatment of cancer. The treatment of cancer must include the benefits of botanical medicines. There are many classes of plant-derived cytotoxic natural products and the structural modification studies for further improvement and development of drug. New anticancer drugs derived from research on plant antitumor agents will be continuously discovered. The activities of flavonoids and the synergistic action shown by them with other drugs make them ideal in alternative cancer therapies. The chemopreventive effects that most flavonoids exert are likely to be the sum of their effect on several distinct mechanisms working inside the cell. The flavonoids have been focused for the research since 1930’s but many of them have been used in traditional medicines for thousands of years in eastern countries. Anthraquinones are an important group of bioactive components found in many species of medicinal herbs such as rhubarb, senna, aloe and purslane. Induction of apoptosis is commonly reported among emodin and aloe-emodin, which involve disruption of mitochondria membrane potential, cytochrome c release, and activation of caspase 3. Emodin and aloe-emodin were also able to induce cell-cycle arrest, involving an increase in p53 expression level and accompanied by upregulation of p21. This suggests that emodin could be a promising candidature for the research and development of new anti-tumor drugs.
Effects of Coptis extract combined with chemotherapeutic agents on ROS production, multidrug resistance, and cell growth in A549 human lung cancer cells
Chengwei He, Rong Rong, Jing Liu, Jianbo Wan, Keyuan Zhou, Jing X Kang,
Abstract Background Non–small cell lung cancer is associated with high expression of multidrug resistance (MDR) proteins and low production of reactive oxygen species (ROS). Coptis extract (COP), a Chinese medicinal herb, and its major constituent, berberine (BER), have anticancer properties. This study aims to investigate the effects of COP and BER combined with chemotherapeutic agents, including fluorouracil (5-FU), camptothecin (CPT), and paclitaxel (TAX) on cell proliferation, ROS production, and MDR in A549 human non-small cell lung cancer cells.
Methods A549 cells were treated with different doses of COP and BER, combined with 5-FU, CPT, and TAX. Cell viability was measured by an XTT (2,3-bis-(2-methoxy-4- nitro-5- sulfophenyl)-2 H-tetrazolium-5-carboxanilide) assay. Intracellular ROS levels were determined by measuring the oxidative conversion of cell permeable 2′,7′-dichlorofluorescein diacetate to fluorescent dichlorofluorescein. MDR of A549 cells was assessed by rhodamine 123 retention assay.
Results Both COP and BER significantly inhibited A549 cell growth in a dose-dependent manner. Combinations of COP or BER with chemotherapeutic agents (5-FU, CPT, and TAX) exhibited a stronger inhibitory effect on A549 cell growth. In addition, COP and BER increased ROS production and reduced MDR in A549 cells.
Conclusion As potential adjuvants to chemotherapy for non–small cell lung cancer, COP and BER increase ROS production, reduce MDR, and enhance the inhibitory effects of chemotherapeutic agents on A549 cell growth.
Anticancer Potentials of Root Extract of Polygala senega and Its PLGA Nanoparticles-Encapsulated Form Saili Paul,1 Soumya Sundar Bhattacharyya,1 Naoual Boujedaini,2 and Anisur Rahman Khuda-Bukhsh1
1Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India 2Boiron Laboratory, 69110 Lyon, France
Abstract Ethanolic extract of Polygala senega (EEPS) had little or no cytotoxic effects on normal lung cells, but caused cell death and apoptosis to lung cancer cell line A549. In the present paper, ethanolic root extract of P. senega (EEPS) was nanoencapsulated (size: 147.7 nm) by deploying a biodegradable poly-(lactic-co-glycolic) acid (PLGA). The small size of the NEEPS resulted in an enhanced cellular entry and greater bioavailability. The growth of cancer cells was inhibited better by NEEPS than EEPS. Both EEPS and NEEPS induced apoptosis of A549 cells, which was associated with decreased expression of survivin, PCNA mRNA, and increased expression of caspase-3, p53 mRNAs of A549 cells. The results show that the anticancer potential of the formulation of EEPS-loaded PLGA nanoparticles was more effective than EEPS per se, probably due to more aqueous dispersion after nanoencapsulation. Therefore, nanoencapsulated ethanolic root extract of P. senega may serve as a potential chemopreventive agent against lung cancer.
Conclusion In conclusion, it has been demonstrated that homeopathic mother tincture of P. senega has an anticancer effect against lung cancer cells in vitro, and PLGA encapsulation helps it to enhance its cellular uptake and anticancer potentials, presumably by increasing drug bioavailability. This should stimulate further research on nano-encapsulation of homeopathic mother tinctures, as also medicinal herbal extracts, particularly with suspected anticancer potentials, to examine whether this would prove to be a novel approach for accelerating anticancer potentials for other cases as well.
Source : Evidence-Based Complementary and Alternative Medicine Volume 2011 (2011), Article ID 517204, 13 pages doi:10.1155/2011/517204 Link to Full Article
Silibinin modulates TNF-α and IFN-γ mediated signaling to regulate COX2 and iNOS expression in tumorigenic mouse lung epithelial LM2 cells
Lori D. Dwyer-Nield1,2,
Rana P. Singh1,3,
Alvin M. Malkinson1,2,
Abstract Silibinin inhibits mouse lung tumorigenesis in part by targeting tumor microenvironment. Tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) can be pro- or anti-tumorigenic, but in lung cancer cell lines they induce pro-inflammatory enzymes cyclooxygenase 2 (COX2) and inducible nitric oxide synthase (iNOS). Accordingly, here we examined mechanism of silibinin action on TNF-α + IFN-γ (hereafter referred as cytokine mixture) elicited signaling in tumor-derived mouse lung epithelial LM2 cells. Both signal transducers and activators of the transcription (STAT)3 (tyr705 and ser727) and STAT1 (tyr701) were activated within 15 min of cytokine mixture exposure, while STAT1 (ser727) activated after 3 h. Cytokine mixture also activated Erk1/2 and caused an increase in both COX2 and iNOS levels. Pretreatment of cells with a MEK, NF-κB, and/or epidermal growth factor receptor (EGFR) inhibitor inhibited cytokine mixture-induced activation of Erk1/2, NF-κB, or EGFR, respectively, and strongly decreased phosphorylation of STAT3 and STAT1 and expression of COX2 and iNOS. Also, janus family kinases (JAK)1 and JAK2 inhibitors specifically decreased cytokine-induced iNOS expression, suggesting possible roles of JAK1, JAK2, Erk1/2, NF-κB, and EGFR in cytokine mixture-caused induction of COX2 and iNOS expression via STAT3/STAT1 activation in LM2 cells. Importantly, silibinin pretreatment inhibited cytokine mixture-induced phosphorylation of STAT3, STAT1, and Erk1/2, NF-κB-DNA binding, and expression of COX2, iNOS, matrix metalloproteinases (MMP)2, and MMP9, which was mediated through impairment of STAT3 and STAT1 nuclear localization. Silibinin also inhibited cytokine mixture-induced migration of LM2 cells. Together, we showed that STAT3 and STAT1 could be valuable chemopreventive and therapeutic targets within the lung tumor microenvironment in addition to being targets within tumor itself, and that silibinin inhibits their activation as a plausible mechanism of its efficacy against lung cancer.
NB Silibinin (INN), also known as silybin, is the major active constituent of silymarin, standardized extract of the milk thistle seeds
Terpinen-4-ol Induces Apoptosis in Human Nonsmall Cell Lung Cancer In Vitro and In Vivo Chieh-Shan Wu,1 Yun-Ju Chen,2 Jeremy J. W. Chen,3 Jeng-Jer Shieh,3 Chia-Hsin Huang,4 Pei-Shan Lin,3 Gee-Chen Chang,3,5,6 JingHua-Tsai Chang,7 and Chi-Chen Lin3,6
1Department of Dermatology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan 2Department of Child Care, College of Humanities and Social Sciences, Southern Taiwan University, Tainan 71005, Taiwan 3Institute of Biomedical Sciences, College of Life Science, National Chung Hsing University, Taichung 40227, Taiwan 4Agricultural Research Institute, Council of Agriculture Executive, Yuan 10014, Taiwan 5Department of Internal Medicine, Division of Chest Medicine, Taichung Veterans General Hospital, Taichung 40705, Taiwan 6Department of Medical Education and Research, Taichung-Veterans General Hospital, Taichung 40705, Taiwan 7Institute of Medicine, Chung Shan Medical University, Taichung 40201,
.Abstract Terpinen-4-ol, a monoterpene component of the essential oils of several aromatic plants, exhibits antitumor effects. In this study, the antitumor effects of terpinen-4-ol and the cellular and molecular mechanisms responsible for it were evaluated and studied, respectively on human nonsmall cell lung cancer (NSCLC) cells. Our results indicated that terpinen-4-ol elicited a dose-dependent cytotoxic effect, as determined by MTT assay. Increased sub-G1 population and annexin-V binding, activation of caspases 9 and 3, cleavage of poly(ADPribose) polymerase (PARP), and a decrease of mitochondrial membrane potential (MMP) indicated involvement of the mitochondrial apoptotic pathway in terpinen-4-ol-treated A549 and CL1-0 cells. Elevation of the Bax/Bcl-2 ratio and a decrease in IAP family proteins XIAP and survivin were also observed following terpinen-4-ol treatment. Notably, terpinen-4-ol was able to increase p53 levels in A549 and CL1-0 cells. Diminution of p53 by RNA interference induced necrosis instead of apoptosis in A549 cells following terpinen-4-ol treatment, indicating that terpinen-4-ol-elicited apoptosis is p53-dependent. Moreover, intratumoral administration of terpinen-4-ol significantly suppressed the growth of s.c. A549 xenografts by inducing apoptosis, as confirmed by TUNEL assay. Collectively, these data provide insight into the molecular mechanisms underlying terpinen-4-ol-induced apoptosis in NSCLC cells, rendering this compound a potential anticancer drug for NSCLC.
Source : Evidence-Based Complementary and Alternative Medicine Volume 2012 (2012), Article ID 818261, 13 pages doi:10.1155/2012/818261 Link to Full Article
Anticancer potentials of root extract of Polygala senega against benzo［a］pyrene-induced lung cancer in mice 1. Saili Paul (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India ) 2. Soumya Sundar Bhattacharyya (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India ) 3. Asmita Samaddar (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235, India ) 4. Naoual Boujedaini (Boiron Laboratory, 20, rue de la Liberation, Sainte-Les-Foy-Lyon, France ) 5. Anisur Rahman Khuda-Bukhsh (Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, Kalyani 741235,
Objective: To evaluate anticancer potentials of Polygala senega on lung cancer induced by benzo［a］pyrene (B［a］P) in mice.
Methods: Swiss albino mice were divided into five groups with each containing six animals. Group 1 served as control, and the animals received olive oil as vehicle. Group 2 animals were treated with B［a］P (50 mg/kg body weight dissolved in olive oil) orally twice a week for four consecutive weeks. Group 3 animals were fed B［a］P as in group 2 and 48% alcohol (since the vehicle of the remedy was alcohol). Group 4 animals were B［a］P-intoxicated mice (as in group 2) which were additionally fed ethanolic extract of Polygala senega (EEPS) daily for 16 weeks. EEPS treatment started after the first dose of B［a］P. Group 5 animals were treated with EEPS alone for 16 weeks to test cytotoxicity of EEPS if any. Mice were sacrificed after 16 weeks and the following parameters were assessed: the anti-oxidant activity measured by 2,2-diphenyl-1-picrylhydrazyl free radical assay, tumor incidence, lung weight and body weight, DNA damage evaluation by comet assay and enzyme-linked immunosorbent assay (ELISA); toxicity biomarkers like catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, lipid peroxidation (LPO) and total thiol content were also detected.
Results: Treatment with EEPS increased the final body weight and significantly decreased the lung weight in group 4 mice (P<0.01) compared with group 3 mice. Comet assay showed that EEPS-treated mice in group 4 presented a decrease of DNA damage significantly (P<0.01) in lung tissues. There was a significant increase observed in the level of p53 in group 4 as compared with group 3 (P<0.01) detected by ELISA. A highly significant increase in tissue LPO with concomitant decrease in the activity of anti-oxidants was observed in group 2 and group 3 mice (P<0.05) compared with the control mice. These adverse changes were reversed significantly in group 4 mice (P<0.01).
Conclusion: Chemopreventive potentials of Polygala senega against chemically induced lung cancer in mice are confirmed.
In vitro and in vivo antitumor activity of Macrothelypteris torresiana and its acute/subacute oral toxicity by X.H. Huang, P.C. Xiong, C.M. Xiong, Y.L. Cai, A.H. Wei, J.P. Wang, X.F. Liang, J.L. Ruan
The aim of this study was to evaluate the antitumor potential of Macrothelypteris torresiana by studying in vitro antitumor activity of the protoapigenone, as well as in vivo antitumor activity and acute/subacute oral toxicity of the total flavonoid fraction from the roots of M. torresiana. Considering that the protoapigenone is a main constituent of the total flavonoid fraction and it might play a key role in the antitumor activity of M. torresiana, the MTT assay was used to investigate the in vitro antitumor activity of the protoapigenone. Our study revealed that the protoapigenone of M. torresiana showed significant antitumor activity towards Hep G2, Tca-8113, MCF-7, M5 and K562 with [IC.sub.50] values of 2.3, 0.6, 0.8, 0.3 and 0.9 [mu]g/ml, respectively. The antitumor potential of the total flavonoid fraction was evaluated using preparations 1, 2 and 3, which were prepared by total flavonoid fraction directly diluted with sterile saline, dissolved using sodium carboxymethyl cellulose (CMC-Na) and included by hydroxypropyl-[beta]-cyclodextrin, respectively. These were investigated in vivo using mouse sarcoma S-180 in BALB/c mice after completing tumor inoculation for 24 h. Pronounced antitumor activity was observed in the treated groups for preparations 2 and 3, and the high and medium doses in particular showed very high inhibition ratio of tumor growth (<50%). No significant difference was observed when compared to the positive control group (5-fluorouracil). The acute/subacute oral toxicity test was performed, and the results of acute oral toxicity showed that the [LD.sub.50] values of preparations 2 and 3 were 2.76 and 0.87 g/kg body wt., respectively. According to the results of the subacute oral toxicity study, the total flavonoid fraction had low toxicity. The overall results of this study suggest that the total flavonoid fraction from the roots of M. torresiana shows significant antitumor activity and represents a potential source of medicine for the treatment of cancer.
NOTE Macrothelypteris torresiana (Gaud.) Ching (Thelypteridaceae) is widely distributed in the south of China and has been used as folk medicine mainly for the treatment of diseases such as hydropsy and traumatic bleeding (Institute of Botany, 1976; Ding, 1982). This recent scientific attention was because of some unusual flavonoids isolated from the species M. torresiana showing significant antitumor activities. Among of them, a unique flavonoid named protoapigenone showed significant antitumor activity against human cancer cell lines Hep G2, Hep 3B (liver), MCF-7 (breast), A549 (lung) and MDA-MB-231 with [IC.sub.50] values of 1.60, 0.23, 0.78, 3.88 and 0.27 ([mu]g/ml, respectively (Lin et al., 2005).
1Department of Life Science, Fu-Jen Catholic University, Taipei 24205, Taiwan 2Department of Internal Medicine, Division of Cardiology, Tian-Sheng Memorial Hospital, Pingtung 92843, Taiwan 3Multi Chemical Laboratory, SGS Taiwan Ltd., Taipei 24803, Taiwan 4Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung City 40201, Taiwan 5Graduate Institute of Cancer Biology, China Medical University and Hospital, Taichung City 40402, Taiwan 6Clinical Laboratory, Chung Shan Medical University Hospital, Taichung City 40201, Taiwan 7Institutes of Biochemistry and Biotechnology, Chung Shan Medical University, No. 110, Sec. 1, Jianguo N. Road, Taichung 40201, Taiwan
Abstract Ocimum gratissimum (OG) is widely used as a traditional herb for its antibacterial activity in Taiwan. Recently, antitumor effect of OG on breast cancer cell is also reported; however, the effects of OG on human pulmonary adenocarcinoma cell A549 remain unclear. Therefore, we aimed to investigate whether aqueous OG extract (OGE) affects viability of A549 cells and the signals induced by OGE in A549 cells. Cell viability assays revealed that OGE significantly and dose-dependently decreased the viability of A549 cell but not that of BEAS-2B cell. Morphological examination and DAPI staining indicated that OGE induced cell shrinkage and DNA condensation for A549 cells. Further investigation showed that OGE enhanced activation of caspase-3, caspase-9 and caspase-8 and increased protein level of Apaf-1 and Bak, but diminished the level of Bcl-2. Additionally, OGE inhibited the phosphorylation of extracellular signal-regulated kinase (ERK) yet enhanced the phosphorylation of c-Jun N-terminal kinase (JNK) and p38 MAP kinase (p38). In conclusion, our findings indicate that OGE suppressed the cell viability of A549 cells, which may result from the activation of apoptotic signaling and the inhibition of anti-apoptotic signaling, suggesting that OGE might be beneficial to lung carcinoma treatment.
Conclusion ....In conclusion, the present study provides evidences that OGE treatments significantly alter viability of lung adenocarcinoma A549 cells through a synergy of induction of apoptotic signaling and suppression of antiapoptotic signaling, as shown in Figure 7. Moreover, OGE treatment simultaneously inhibits the activation of ERK and enhances the activation of JNK and p38, which is consistent with the enhanced apoptotic signaling and reduced antiapoptotic signaling based on their well-known effects on these signal cascades (Figure 7). By manipulating both arms of apoptotic and antiapoptotic pathway, OGE represents a promisingly effective chemopreventive agent against lung adenocarcinoma.
Source :Evidence-Based Complementary and Alternative Medicine Volume 2011 (2011), Article ID 739093, 7 pages doi:10.1155/2011/739093 Link to Full Article
Growth Inhibition and Regression of Lung Tumors by Silibinin: Modulation of Angiogenesis by Macrophage-Associated Cytokines and Nuclear Factor-κB and Signal Transducers and Activators of Transcription 3
Abstract The latency period for lung tumor progression offers a window of opportunity for therapeutic intervention. Herein, we studied the effect of oral silibinin (742 mg/kg body weight, 5 d/wk for 10 weeks) on the growth and progression of established lung adenocarcinomas in A/J mice. Silibinin strongly decreased both tumor number and tumor size, an antitumor effect that correlates with reduced antiangiogenic activity. Silibinin reduced microvessel size (50%, P < 0.01) with no change in the number of tumor microvessels and reduced (by 30%, P < 0.05) the formation of nestin-positive microvessels in tumors. Analysis of several proteins involved in new blood vessel formation showed that silibinin decreased the tumor expression of interleukin-13 (47%) and tumor necrosis factor-α (47%), and increased tissue inhibitor of metalloproteinase-1 (2-fold) and tissue inhibitor of metalloproteinase-2 (7-fold) expression, without significant changes in vascular endothelial growth factor levels. Hypoxia- inducible factor-1α expression and nuclear localization were also decreased by silibinin treatment. Cytokines secreted by tumor cells and tumor-associated macrophages regulate angiogenesis by activating nuclear factor-κB (NF-κB) and signal transducers and activators of transcription (STAT). Silibinin decreased the phosphorylation of p65NF-κB (ser276, 38%; P < 0.01) and STAT-3 (ser727, 16%; P < 0.01) in tumor cells and decreased the lung macrophage population. Angiopoietin-2 (Ang-2) and Ang-receptor tyrosine kinase (Tie-2) expression were increased by silibinin. Therapeutic efficacy of silibinin in lung tumor growth inhibition and regression by antiangiogenic mechanisms seem to be mediated by decreased tumor-associated macrophages and cytokines, inhibition of hypoxia-inducible factor-1α, NF-κB, and STAT-3 activation, and up-regulation of the angiogenic inhibitors, Ang-2 and Tie-2.
MORE: Lung cancer is the leading cause of cancer death in both men and women in the United States, with an estimated 213,380 new lung cancer cases and 160,390 associated deaths in 2007 (1). The 5-year survival rate of 14% has shown little improvement over the last 30 years, even with the development of molecularly targeted therapies such as epidermal growth factor receptor inhibitors. Tobacco exposure has been implicated in 90% of lung carcinomas; compared with never smokers, smokers have a 20-fold greater risk of developing lung cancer (2). Because smoking is the major risk factor for developing lung cancer and most smokers have small pulmonary nodules, strategies for inducing nodule regression or preventing their further growth should decrease the number of patients diagnosed with advanced malignant disease.
Efforts are being made towards identifying dietary supplements to prevent and treat lung cancer. One such agent, silibinin, inhibits the growth of various cancer cell lines and primary tumors in several chemically induced rodent models, including mouse lung (3–7). Silibinin is a flavonolignan, a major component in the silymarin complex of flavonolignans and polyphenols present in milk thistle (Silybum marianum) seeds. Silymarin has been extensively used in patients with liver disease for decades (8). Silibinin has shown strong anticancer efficacy against SHP-77 and A549 lung cancer cells, in which it inhibits cell growth and induces cell cycle arrest (9). It also inhibits the invasion of lung cancer cells via down-regulating phosphoinositide 3-kinase-Akt and mitogen-activated protein kinase signaling pathways and decreased production of urokinase-plasminogen activator and matrix metalloproteinase-2 (10, 11). Silibinin inhibits the in vivo growth of A549 xenografts in nude mice, reduces systemic toxicity of doxorubicin, and reduces doxorubicin-induced chemoresistance by inhibiting nuclear factor-κB (NF-κB) signaling (12)...........
In summary, oral silibinin showed antitumor effects in urethane-induced and established lung adenocarcinomas, most likely by decreasing microvessel size and inhibiting newly formed microvessel growth in tumors. The decrease in TAM infiltration into lungs as well as the lower levels of angiogenic cytokines, and greater TIMP-1 and TIMP-2 concentrations, along with the inhibition of HIF-1α, NF-κB, and STAT3 activation, could account for the antiangiogenic effects of silibinin. Additionally, elevating levels of Ang-2 and Tie-2 without changing VEGF amounts could have led to microvessel regression in tumors by silibinin. Overall, our findings here, together with our earlier studies (7), suggest that silibinin is a promising agent for intervention in human lung cancer oncogenesis.
Grape Seed Proanthocyanidins Inhibit the Growth of Human Non-Small Cell Lung Cancer Xenografts by Targeting Insulin-Like Growth Factor Binding Protein-3, Tumor Cell Proliferation, and Angiogenic Factors
Abstract Purpose: Lung cancer is a leading cause of cancer-related deaths worldwide. Here, we assessed the chemotherapeutic effect of grape seed proanthocyanidins (GSPs) on human non-small cell lung cancer (NSCLC) cells in vitro and in vivo using a tumor xenograft model.
Experimental Design: The effects of GSPs on human NSCLC cell lines in terms of cellular proliferation were determined. The chemotherapeutic effects of a GSP- supplemented AIN76A control diet fed to nude mice bearing tumor xenografts (A549 and H1299) were evaluated in terms of biomarkers of cell proliferation and angiogenesis and on insulin-like growth factor binding protein-3 using immunohistochemical detection, ELISA, and Western blotting.
Results:In vitro treatment of NSCLC cells with GSPs resulted in inhibition of cellular proliferation. Administration of GSPs (0.1%, 0.2%, and 0.5%, w/w) as a supplement of an AIN76A control diet resulted in a dose-dependent inhibition of the growth of NSCLC (A549 and H1299) tumor xenografts in athymic nude mice (25-76%; P < 0.05-0.001). The growth-inhibitory effect of GSPs on the NSCLC xenograft tumors was associated with the enhancement of the levels of insulin-like growth factor binding protein-3 in the tumor microenvironment and plasma and antiproliferative, antiangiogenic, and proapoptotic effects.
Conclusions:This preclinical study reveals for the first time that dietary GSPs have the ability to inhibit the growth of human NSCLC tumor xenografts grown in vivo in athymic nude mice. More studies are needed to develop GSPs as a pharmacologically safe agent for the prevention of lung cancer in humans.
SOURCE:doi: 10.1158/1078-0432.CCR-08-1901 Clinical Cancer Research February 2009 15; 821 LINK TO FULL ARTICLE
Abstract Background Inverse associations between cruciferous vegetable intake and lung cancer risk have been consistently reported. However, associations within smoking status subgroups have not been consistently addressed.
Methods We conducted a hospital-based case-control study with lung cancer cases and controls matched on smoking status, and further adjusted for smoking status, duration, and intensity in the multivariate models. A total of 948 cases and 1743 controls were included in the analysis.
Results Inverse linear trends were observed between intake of fruits, total vegetables, and cruciferous vegetables and risk of lung cancer (ORs ranged from 0.53-0.70, with P for trend < 0.05). Interestingly, significant associations were observed for intake of fruits and total vegetables with lung cancer among never smokers. Conversely, significant inverse associations with cruciferous vegetable intake were observed primarily among smokers, in particular former smokers, although significant interactions were not detected between smoking and intake of any food group. Of four lung cancer histological subtypes, significant inverse associations were observed primarily among patients with squamous or small cell carcinoma - the two subtypes more strongly associated with heavy smoking.
Conclusions Our findings are consistent with the smoking-related carcinogen-modulating effect of isothiocyanates, a group of phytochemicals uniquely present in cruciferous vegetables. Our data support consumption of a diet rich in cruciferous vegetables may reduce the risk of lung cancer among smokers.
Effects of Feiyanning Decoction on gene expression of nuclear factor-κB activated by tumor necrosis factor-α in lung adenocarcinoma cell line
1. Ju-yong WANG (Tumor Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China 2. Ye JIN (Department Bioengineering, School of Life Sciences, Shanghai University, Shanghai 200444, China ) 3. Zhen-ye XU (Tumor Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China ) 4. Zhan ZHENG (Tumor Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China )
Objective: To study the effects of Feiyanning Decoction, a compound traditional Chinese herbal medicine, on gene expression of nuclear factor-κB (NF-κB) activated by tumor necrosis factor-α (TNF-α) in lung adenocarcinoma cell line (A549).
Methods: A549 cells were incubated with rat serum containing Feiyanning at different concentrations for 24 and 48 h, respectively. Morphology of cells was observed by an inverted microscope after treatment with reagents. The cell proliferation was examined by 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2, 4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-8) assay. The expressions of NF-κB and inhibitor κBα (IκBα) were studied by Western blotting. NF-κB-dependent luciferase reporter (3×κB-luc) was transfected for 24 h, and the cells were treated with the reagents for 24 h, and then the transcriptional activity of NF-κB promoter was detected by luciferase assay.
Results: TNF-α (1 μg/L) strongly induced the expression of NF-κB by approximately 1.76-fold compared with the control in the nuclei of A549 cells, and the induced NF-κB expression was significantly suppressed by addition of Feiyanning (P<0.01). In addition, Feiyanning inhibited the transcriptional activity of the NF-κB promoter. However, we observed no significant changes in IκBα expression (P>0.05).
Conclusion: Feiyanning Decoction can markedly inhibit human lung cancer A549 cell proliferation, which may be partly due to inhibition of NF-κB activation induced by TNF-α. It is therefore expected to be a new strategy for treating lung cancer.
Cancer Chemopreventive Potential of Apples, Apple Juice, and Apple Components Author Clarissa Gerhauser
Affiliation Division of Toxicology and Cancer Risk Factors, German Cancer Research Center (DKFZ), Heidelberg, Germany Review Abstract
Apples (Malus sp., Rosaceae) are a rich source of nutrient as well as non-nutrient components and contain high levels of polyphenols and other phytochemicals. Main structural classes of apple constituents include hydroxycinnamic acids, dihydrochalcones,flavonols (quercetin glycosides), catechins and oligomeric procyanidins, as well as triterpenoids in apple peel and anthocyanins in red apples. Several lines of evidence suggest that apples and apple products possess a wide range of biological activities which may contribute to health beneficial effects against cardiovascular disease, asthma and pulmonary dysfunction, diabetes, obesity, and cancer (reviewed by Boyer and Liu, Nutr J 2004). The present review will summarize the current knowledge on potential cancer preventive effects of apples, apple juice and apple extracts (jointly designated as apple products). In brief, apple extracts and components, especially oligomeric procyanidins, have been shown to influence multiple mechanisms relevant for cancer prevention in in vitro studies. These include antimutagenic activity, modulation of carcinogen metabolism, antioxidant activity, anti-inflammatory mechanisms, modulation of signal transduction pathways, antiproliferative and apoptosis-inducing activity, as well as novel mechanisms on epigenetic events and innate immunity. Apple products have been shown to prevent skin, mammary and colon carcinogenesis in animal models. Epidemiological observations indicate that regular consumption of one or moreapples a day may reduce the risk for lungand colon cancer.