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Anticancer Effect of Fraction Isolated from Medicinal Birch Polypore Mushroom, Piptoporus betulinus (Bull.: Fr.) P. Karst. (Aphyllophoromycetideae): In Vitro Studies
Abstract
Piptoporus betulinus (Bull.: Fr.) P. Karst. (Fomitopsidaceae) has been commonly used in folk medicine as an antiparasitic and antimicrobial agent in the treatment of wounds and for the treatment of rectal cancer and stomach diseases. Tea obtained from this mushroom has antibacterial, antifatiguing, immunoenhancing, and soothing properties. The purpose of the present study was the evaluation of in vitro anticancer activity of fraction isolated from P. betulinus. The effect on cell proliferation, motility, and viability was assessed in a range of cancer and normal cells. P. betulinus fraction prepared from dried fruiting bodies was subjected to anticancer evaluation in human lung carcinoma (A549), colon adenocarcinoma (HT-29), and rat glioma (C6) cell cultures. Human skin fibroblasts (HSF), bovine aorta endothelial cells (BAEC), models of rat oligodenrocytes (OLN-93), hepatocytes (Fao), rat astroglia, and mouse neurons (P19) were applied to test toxicity in normal cells. The following methods were applied: tumor cell proliferation (MTT assay and BrdU assay), cytotoxicity (LDH assay), tumor cell motility (wound assay), tumor cell morphology (May-Grünwald-Giemsa staining), and death detection (ELISA). P. betulinus fraction elicited anticancer effects that were attributed to decreased tumor cell proliferation, motility, and the induction of morphological changes. Of note is the fact that it produced no or low toxicity in tested normal cells.
... The effect of the biologically active compounds extracted from Piptoporus betulinus with the use of the different solvents on cancer cell viability has been shown previously [9,11,36]. A549 human lung carcinoma, C6 rat glioma, and HT-29 human colon adenocarcinoma cell viability was significantly affected by Piptoporus betulinus ethyl acetate fraction in a dose-response manner, as was shown by MTT assay [36]. ...
... The effect of the biologically active compounds extracted from Piptoporus betulinus with the use of the different solvents on cancer cell viability has been shown previously [9,11,36]. A549 human lung carcinoma, C6 rat glioma, and HT-29 human colon adenocarcinoma cell viability was significantly affected by Piptoporus betulinus ethyl acetate fraction in a dose-response manner, as was shown by MTT assay [36]. ...
... The measurement of LDH release from the cells, damaged by the active compounds present in Piptoporus betulinus extract, showed a statistically significant increase in LDH in the culture medium collected from above the cells treated by the highest concentration of the extract (10 µL mL A cell-specific cytotoxic effect of Piptoporus betulinus extract was demonstrated in the previous study by Lemieszek et al. [36]. Piptoporus betulinus ethyl acetate fraction, used in the highest concentration (250 µg mL −1 ), had statistically significant cytotoxic activity in an LDH assay against three rat cancer cell lines, FAO hepatoma, OLN-93 oligodendroglial, and astroglia, while two normal cell lines, HSF human skin fibroblasts and BAEC bovine aorta endothelial cells, were not sensitive to the action of this fraction [36]. ...
Piptoporus betulinus is a fungus known for its medicinal properties. It possesses antimicrobial, anti-inflammatory, and anti-cancer activity. In this study, several tests were performed to evaluate the cytotoxic effect of the ethanolic extract of Piptoporus betulinus on two melanoma human cell lines, WM115 primary and A375 metastatic cell lines, as well as Hs27 human skin fibroblasts. The extract proved to affect cancer cells in a dose-dependent manner, and at the same time showed a low cytotoxicity towards the normal cells. The total phenolic content (TPC) was determined spectrophotometrically by the Folin-Ciocalteu method (F-C), and the potential antioxidant activity was measured by ferric-reducing antioxidant power (FRAP) assay. One of the active compounds in the extract is betulin. It was isolated and then its cytotoxic activity was compared to the results obtained from the Piptoporus betulinus extract. To further understand the mechanism of action of the extract’s anticancer activity, tests on model cell membranes were conducted. A model membrane of a melanoma cell was designed and consisted of 1,2-dimyristoyl-sn-glycero-3-phosphocholine, disialoganglioside-GD1a and cholesterol: DMPC:GD1a:chol (5:2:3 mole ratio). Changes in a Langmuir monolayer were observed and described based on Π-Amol isotherm and compressibility modulus changes. LB lipid bilayers were deposited on a hydrophilic gold substrate and analyzed by IR and X-ray photoelectron spectroscopy. Our study provides new data on the effect of Piptoporus betulinus extract on melanoma cells and its impact on the model of melanoma plasma membranes.
... A fraction prepared from dried fruiting bodies was subjected to anticancer evaluation in human lung carcinoma (A549), colon adenocarcinoma (HT-29), and rat glioma (C6) cell cultures, and elicited cytotoxic effects that were attributed to decreased tumor cell proliferation, motility and induction of morphological changes. Moreover, it produced no or low toxicity in tested normal cells 43 . ...
Zeyad Alresly
The aim of the present dissertation was to investigate the biological and chemical potential of two European mushroom species: Fomitopsis betulina and Calvatia gigantea. For this purpose, different extracts of both fungi were tested for: antimicrobial, antifungal, cytotoxic, in vitro wound healing, and anti-adhesive properties. Bioassay-guided fractionation led to the isolation of bioactive compounds, altogether 20 compounds were isolated and identified. The compounds were obtained from the ethyl acetate extracts, they included triterpenes, sterols and aromatic compounds. The separated substances from both fungi were proved for biological activities, some of them showed antimicrobial and cytotoxic activities.
... Dried fruiting bodies fraction of P. betulinus was evaluated for their anti-cancer properties and found to have an effect against human lung carcinoma (A549), colon adenocarcinoma (HT-29), and rat glioma (C6). This fraction showed decrease in the proliferation of tumor cells along with induction of morphological changes in cancer cell lines (Lemieszek et al. 2009). Phellinus baumii is a folk medicinal mushroom also called as oak bracket has cytotoxic activity against human melanoma cells A375 (Yang et al. 2020). ...
Today mankind confronts a heap of challenges for survival due to the advent of health-related issues, drug resistances, and imbalances in the ecosystems. In the era of technology, man has perpetually been endeavoring to search for diverse biotic components that can potentially be addressing the complicated life troubling issues. In this context, the fungi in general and mushrooms in particular have played an indispensable role in protecting and curing various health problems. Macrofungi or mushrooms are contemplated as biological and genetic resources with high nutritional, medicinal, and biotechnological potential. The interest in mushrooms has cultivated momentously in the last few decades, being promoted by the discovery of a repertoire of chemically disparate biologically active compounds having biopharmaceutical applications arbitrated through defined mechanisms (anti-tumor, anti-inflammatory, anti-cancer, anti-oxidative hepatoprotective, anti-viral, immunomodulating hypocholesterolemic, and anti-bacterial). The escalating knowledge about chemistry, biotechnology, and molecular biology of mushrooms as well as an improvement in screening methods has led to rapid surge in the application of mushrooms for medicinal purposes which in turn, have galvanized the development of several novel mycopharmaceuticals based on mushroom bioprospection. Taking into consideration the importance of mushrooms, this chapter aims to zero in on the nutritive value, functionalities of mushrooms, and potential applications in food industry.
... In spite of UV-Vis and MS spectra indicating the presence of various metabolites in the single fractions a comparison with databases of known fungal substances did only result in the identification of one single compound being 4-methoxybenzaldehyde. Although this substance is not very interesting from a pharmaceutical point of view the findings are in agreement with data reported by Lemieszek (Lemieszek et al. 2009). Surprisingly in case of the extract of P. betulinus none of the allegedly characteristic triterpenoids (Peintner, R.;Pöder, U.;Pümpel 1998;Hybelhauerová et al. 2008;Alresly et al. 2016) were found. ...
This thesis demonstrates the pharmaceutical potential of aqueous extracts of European polypores as well as their effects on the inflammatory response and their anti-thrombogenic properties. Furthermore the feasibility of producing an enzyme functionalised antibacterial nanofibrous wound dressing containing active constituents of polypores that participate in the healing process is demonstrated.
The use of Fomitopsis betulina (birch mushroom) for the treatment of various ailments and diseases has been known for several thousand years. Triterpenes including polyporenic acids and some further compounds have been isolated as bioactive metabolites. Numerous in-vitro and some in-vivo studies showed, among others, inhibiting effects on cancer cells, immunomodulating and antiinflammatory activities. However, further extensive research and the involvement of the pharmaceutical industry are required to exploit this potential not only in the form of food supplements but also as approved drugs.
Background/Objectives: This study investigated the anticancer potential of an aqueous extract of the fungus Fomitopsis betulina. Methods: The study assessed the effect of the extract on nine cancer cell lines, including melanoma (LM-MEL-75), lung cancer (A549), and colorectal cancer (HT29, LoVo), and four normal cell lines. The cytotoxicity of the extract was evaluated using MTT, sulforhodamine-B (SRB), and clonogenic viability assays. Additionally, the study examined the effect of the extract on plant model organisms, garden cress (Lepidium sativum) and common onion (Allium cepa), to further investigate its biological activity. Results: The assays demonstrated selective cytotoxicity of the extract toward cancer cells, while sparing normal cells. The extract induced significant cytotoxic effects at lower concentrations in lung cancer, melanoma, and colon cancer cells, showing promise as a potential anticancer agent. The results also revealed that the extract inhibited seed germination and root growth, suggesting its potential to disrupt cell cycles and induce apoptosis. Conclusions: This study highlights the therapeutic potential of F. betulina and highlights the need for further research to identify the active ingredients and mechanisms underlying its anticancer effects.
Interest in edible and medicinal macrofungi is millennial in terms of their uses in health and food products in Central Asia, while interest in inedible and medicinal macrofungi has grown in popularity in recent years. Edible and inedible medicinal basidiomycetes were collected during field surveys from different regions of Uzbekistan. The morphological characters and similarity assessment of rDNA-Internal Transcribed Spacer sequence data were used to measure diversity and habitat associations. A number of 17 species of medicinal macrofungi of ethnomycological and medicinal interest was found associated with 23 species of trees and shrubs belonging to 11 families and 14 genera. Polyporaceae and Hymenochaetaceae were represented by the highest number of species followed by Ganodermataceae, Fomitopsidaceae, Auriculariaceae, Cerrenaceae, Grifolaceae, Phanerochaetaceae, Laetiporaceae, Schizophyllaceae, and Stereaceae. The highest number of medicinal basidiomycete species was reported in the following host genera: Acer, Betula, Celtis, Crataegus, Juglans, Juniperus, Lonicera, Malus, Morus, Platanus, Populus, Prunus, Quercus, and Salix. An updated list of edible and inedible medicinal mushrooms identified in Uzbekistan, their morphological characteristics, and phylogenetic placement are given for the first time. Information is provided on their uses in traditional and modern medicine. Their bioactive compounds and extracts can be applied as medicines, as well as food and cosmetic ingredients.
While mephedrone (4-methylmethcathinone), a synthetic cathinone derivative, is widely abused by adolescents and young adults, the knowledge about its long-term effects on memory processes is limited. Kynurenic acid (KYNA) is a neuroactive metabolite of the kynurenine pathway of tryptophan degradation. KYNA is considered an important endogenous modulator influencing physiological and pathological processes, including learning and memory processes. The aim of this study was to determine whether (A) binge-like mephedrone administration (10.0 and 30.0 mg/kg, intraperitoneally, in 4 doses separated by 2 h) induces memory impairments, assessed 2, 8 and 15 days after mephedrone cessation in the passive avoidance test in mice, and whether (B) KYNA is involved in these memory processes. To clarify the role of KYNA in the mephedrone effects, its level in the murine brain in vivo, and in cortical slices in vitro, as well as the activities of kynurenine aminotransferases (KATs) I and II were assessed. Furthermore, cell line experiments were conducted to investigate the effects of mephedrone on normal human brain cells. Our results showed memory impairments 8 and 15 days after binge-like mephedrone administration. At the same time, reduction in the KYNA level in the murine brain was noted. In vitro studies showed no effect of mephedrone on the production of KYNA in cortical slices or on the activity of the KAT I and II enzymes. Finally, exposure of normal cells to mephedrone in vitro resulted in a modest reduction of cell viability and proliferation.
Mushrooms degrade matter to produce metabolite for sustaining life. For degradation they produce enzymes and certain metabolites. Both the enzymes and the metabolites are of use for human. The metabolites have medicinal applications. The traditional use and the scientific work on some traditionally used medicinal mushrooms have been discussed here. Some of the mushrooms are Auricularia delicata, A. polytricha, A. auricular, Agaricus blazei, Coprinus comatus, Cordyceps spp., Fomes fomentarius, Fomitopsis pinicola, Ganoderma lucidum, etc. Auricularia delicata has been used in the traditional medicine of Manipur, India, for dysentery and liver healing therapy. A scientific investigation done by one of the authors on traditionally used Auricularia species showed its hepatoprotective activity. The compound isolated from the ethyl acetate extract was chlorogenic acid. It is known to have hepatoprotective activity. This has been a good example illustrating that traditional medicine is a good lead to drug discovery.
Among the numerous items of equipment with the ‘Iceman’, who died more than 5000 years ago on an alpine glacier, were three fungal objects: two different shaped fruitbody pieces of the polypore Piptoporus betulinus, each mounted separately on a leather thong, and, found in his girdle bag, a relatively large quantity of tinder material prepared from the ‘true tinder bracket’ Fomes fomentarius. A full description of these items and a chronological report on their identification is given. The question about the possible use of the fungi is discussed on the basis of a comprehensive collection of ethnomycological and pharmacological literature data.
Kynurenic acid (KYNA) is a broad-spectrum antagonist at all subtypes of ionotropic glutamate receptors, but is preferentially active at the strychnine-insensitive glycine allosteric site of the N-methyl-D-aspartate (NMDA) receptor and is also a non-competitive antagonist at the alpha7 nicotinic receptor. KYNA occurs in the CNS, urine, serum and amniotic fluid. Whilst it possesses anticonvulsant and neuroprotective properties in the brain, its role in the periphery, however, is unknown. In this study we demonstrated the presence of kynurenine aminotransferase (KAT) I and II in the cytoplasm of bovine aortic endothelial cells (BAEC). BAEC incubated in the presence of the KYNA precursor L-kynurenine synthesized KYNA concentration- and time-dependently. KYNA production was inhibited by the aminotransferase inhibitor aminooxyacetic acid but was not affected by a depolarising concentration of K(+) or by 4-aminopyridine. The glutamate agonists L-aspartate and L-glutamate depressed KYNA production significantly. The selective ionotropic glutamate receptor agonists alpha-amino-2,3-dihydro-5-methyl-3-oxo-4-isoxazolepropionic acid (AMPA) and NMDA were ineffective in this respect. D,L-Homocysteine and L-homocysteine sulphinic acid lowered KYNA production in BAEC. Further investigations are needed to assess the role and importance of KYNA in vessels and peripheral tissues.
Insulin-like growth factors (IGFs) are potent proliferation stimulators for numerous tumor cells and often function as autocrine growth factors. We have previously shown that exogenous IGF-I and IGF-II enhance proliferation of colorectal carcinoma cells. The biological signal of both factors is transmitted through the IGF-I receptor (IGF-I-R). This receptor was expressed in 12/12 colorectal carcinoma cell lines tested. αIR3, a neutralizing monoclonal antibody (MAb) directed against the human IGF-I-R, inhibited proliferation in 7/12 lines (Caco-2, HT-29, LS411N, LSS13, LS1034, WiDr and SW620), as reflected by a reduction of MTT conversion (19 to 42%), a decrease in cell number (39 to 72%) and an increase in doubling time (up to 2-fold). In addition, in 4 cell lines (Caco-2. LSS13, LS1034, WiDr) αIR3 suppressed colony formation in methylcellulose (40 to 84%). Excess of exogenous IGF completely neutralized αIR3-mediated inhibitory effects. Northern blot analysis revealed abundant expression of 2 IGF-II transcripts of 5.0 and 4.3 kb in LS1034 cells. In addition, we observed that growth inhibition by αIR3 was correlated with a more differentiated phenotype. Our results suggest that growth of many colorectal carcinoma cell lines is regulated by autocrine IGF-II-mediated stimulation of the IGF-I-R.
Escape of cancer cells from the circulation (extravasation) is thought to be a major rate-limiting step in metastasis, with few cells being able to extravasate. Furthermore, highly metastatic cells are believed to extravasate more readily than poorly metastatic cells. We assessed in vivo the extravasation ability of highly metastatic ras-transformed NIH 3T3 cells (PAP2) versus control nontumorigenic nontransformed NIH 3T3 cells and primary mouse embryo fibroblasts. Fluorescently labeled cells were injected intravenously into chicken embryo chorioallantoic membrane and analyzed by intravital videomicroscopy. The chorioallantoic membrane is an appropriate model for studying extravasation, since, at the embryonic stage used, the microvasculature exhibits a continuous basement membrane and adult permeability properties. The kinetics of extravasation were assessed by determining whether individual cells (n = 1481) were intravascular, extravascular, or in the process of extravasation, at 3, 6, and 24 h after injection. Contrary to expectations, our results showed that all three cell types extravasated with the same kinetics. By 24 h after injection > 89% of observed cells had completed extravasation from the capillary plexus. After extravasation, individual fibroblasts of all cell types demonstrated preferential migration within the mesenchymal layer toward arterioles, not to venules or lymphatics. Thus in this model and for these cells, extravasation is independent of metastatic ability. This suggests that the ability to extravasate in vivo is not necessarily predictive of subsequent metastasis formation, and that postextravasation events may be key determinants in metastasis.
We identified betulinic acid (BetA) as a new cytotoxic agent active against neuroectodermal tumor cells including neuroblastoma, medulloblastoma, glioblastoma and Ewing's sarcoma cells representing the most common solid tumors of childhood. BetA induced apoptosis independent of wild-type p53 protein and accumulation of death inducing ligand/receptor systems such as CD95. BetA had a direct effect on mitochondria resulting in the release of soluble apoptogenic factors such as cytochrome c or AIF from mitochondria into the cytosol where they induced activation of caspases. Over expression of the anti-apoptotic proteins Bcl-2 or Bcl-XL that blocked loss of the mitochondrial membrane potential and cytochrome c release from mitochondria conferred resistance to BetA at the level of mitochondrial dysfunction, protease activation and nuclear fragmentation. Neuroblastoma cells resistant to CD95- or doxorubicin-triggered apoptosis remained sensitive to treatment with BetA suggesting that BetA may bypass some forms of resistance. Moreover, BetA exhibited potent antitumor activity on primary tumor cell cultures from all neuroblastoma (4/4), all medulloblastoma (4/4) and most glioblastoma patients (20/24) ex vivo. These findings suggest that BetA may be a promising new agent in the treatment of neuroectodermal tumors in vivo.
Abbreviations: AIF, apoptosis inducing factor; BetA, betulinic acid; DiOC6(3), 3,3'-dihexyloxacarbocyanide iodide; DTT, Dithiothreitol; ECL, enhanced chemiluminescence; FACS, fluorescence-activated cell-sorting; ICE, interleukin 1β-converting enzyme; ΔYm mitochondrial transmembrane potential; PARP, poly(ADP-ribose) polymerase; zVAD.fmk, benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone.
Activities of cellulolytic and hemicellulolytic enzymes endo-1,4-beta-glucanase, exo-1,4-beta-glucanase, 1,4-beta-glucosidase, endo-1,4-beta-xylanase, 1,4-beta-xylosidase and 1,4-beta-mannosidase and ligninolytic enzymes Mn-peroxidase and laccase were detected during the growth of the white-rot fungus Pleurotus ostreatus on wheat straw in the presence and absence of cadmium. The loss of substrate dry weight and Mn-peroxidase activity decreased with increasing Cd concentration, whereas the activities of endo-1,4-beta-glucanase, 1,4-beta-glucosidase and laccase were highly increased in the presence of metal. The onset of hemicellulose-degrading enzyme activity was delayed in the presence of cadmium. The degradation of a model synthetic dye Poly B-411 did not correspond to the activities of ligninolytic enzymes. This is the first report about 1,4-beta-mannosidase in P. ostreatus.